WO1998034094A1 - Optical apparatus - Google Patents

Optical apparatus Download PDF

Info

Publication number
WO1998034094A1
WO1998034094A1 PCT/NZ1998/000009 NZ9800009W WO9834094A1 WO 1998034094 A1 WO1998034094 A1 WO 1998034094A1 NZ 9800009 W NZ9800009 W NZ 9800009W WO 9834094 A1 WO9834094 A1 WO 9834094A1
Authority
WO
WIPO (PCT)
Prior art keywords
reflector
flow
electromagnetic radiation
particles
reflective surface
Prior art date
Application number
PCT/NZ1998/000009
Other languages
French (fr)
Inventor
Johnathan Sharpe
Peter Niven Schaare
Original Assignee
The Horticulture & Food Research Institute Of New Zealand Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to CA002279574A priority Critical patent/CA2279574C/en
Priority to NZ337340A priority patent/NZ337340A/en
Application filed by The Horticulture & Food Research Institute Of New Zealand Ltd. filed Critical The Horticulture & Food Research Institute Of New Zealand Ltd.
Priority to EP98901601A priority patent/EP1017987B1/en
Priority to AU57836/98A priority patent/AU752985B2/en
Priority to DE69830598T priority patent/DE69830598T2/en
Priority to US09/355,461 priority patent/US6819411B1/en
Priority to AT98901601T priority patent/ATE298084T1/en
Priority to JP53276398A priority patent/JP4323571B2/en
Publication of WO1998034094A1 publication Critical patent/WO1998034094A1/en
Priority to US10/990,648 priority patent/US7221453B2/en
Priority to US11/805,572 priority patent/US7586604B2/en
Priority to US12/555,641 priority patent/US7929137B2/en
Priority to US12/944,308 priority patent/US8975035B2/en
Priority to US13/086,223 priority patent/US8553226B2/en
Priority to US13/764,640 priority patent/US8610892B2/en
Priority to US14/635,910 priority patent/US20150204772A1/en

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • G01N15/14Electro-optical investigation, e.g. flow cytometers
    • G01N15/1434Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
    • G01N15/1436Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47Scattering, i.e. diffuse reflection
    • G01N21/49Scattering, i.e. diffuse reflection within a body or fluid
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B5/00Optical elements other than lenses
    • G02B5/04Prisms
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • G01N15/14Electro-optical investigation, e.g. flow cytometers
    • G01N15/1434Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • G01N15/14Electro-optical investigation, e.g. flow cytometers
    • G01N15/1456Electro-optical investigation, e.g. flow cytometers without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
    • G01N15/1459Electro-optical investigation, e.g. flow cytometers without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
    • G01N15/149
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • G01N15/14Electro-optical investigation, e.g. flow cytometers
    • G01N15/1434Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
    • G01N2015/1452Adjustment of focus; Alignment
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/645Specially adapted constructive features of fluorimeters
    • G01N2021/6463Optics
    • G01N2021/6469Cavity, e.g. ellipsoid

Definitions

  • This invention relates to an optical apparatus.
  • this invention has application to the field of flow cytometry.
  • inventive aspects have application beyond flow cytometry and may have broad application in the field of optics generally.
  • aspects of the invention may be used in photometry or optical particle detection apparatus.
  • the light source is directed onto the particle from a single direction.
  • An analysis may be made of light reflected or produced by the particle eg. fluorescence to reveal certain properties of the particle.
  • the particular portion of the particle illuminated depends on the orientation of the particle with respect to the light source. Where the particle or object is asymmetrical, the light measurements will vary depending on which portion is illuminated, making it difficult to analyse the particle or object as a whole.
  • Flow cytometers are often used to measure the properties of cells or particles which are carried in a stream of fluid.
  • the stream is generally comprised of a sheath fluid into the centre of which is injected a narrow aqueous suspension of cells/particles.
  • the sheath fluid focuses the sample cells/particles into single file.
  • the stream containing the particles/cells passes through an inspection point which is the focus of an intense light beam.
  • the particles/cells may have been stained with a light - sensitive stain which when illuminated, will absorb the incident light and fluoresce. Light scatters off the particles and/or alternatively causes fluorescence. This scattered or fluorescent light is then measured by a detector generally aligned with the incident beam.
  • the characteristics of the detected signal(s) such as peak intensity, peak area or other characteristics of interest may then be used to derive properties of the particle, for example size.
  • the detected signal(s) may be used to trigger sorting hardware which can be programmed to divert droplets from the stream of fluid.
  • sorting hardware which can be programmed to divert droplets from the stream of fluid.
  • the sorting criteria will vary with the application, for example, the sorting may be conducted according to size or, in the case of spermatozoa, the DNA content of the cell.
  • particle asymmetry often renders the optical characteristics of a particle difficult to measure.
  • a flat particle can pass through the inspection point with a random orientation.
  • the intensity of the resultant scattered or fluorescent light may vary according to particle orientation and the detectors will measure different light intensities at different locations.
  • particle asymmetry can lead to a reduced resolution of measurement of the particles. It follows that, in cytometers with a sorting capability, this reduced resolution in measurement of the particles results in a decreased ability to accurately separate populations of cells with different optical properties. Such a problem is encountered in separation of male and female mammalian sperm.
  • sperm carry the sex determining chromosomes and the total DNA content found in male and female sperm may differ.
  • the difference in the DNA content between male and female sperm is approximately 4%.
  • This difference in DNA provides a means by which sperm may be separated in a sorting flow cytometer, making a predetermination of an offspring's sex possible when artificial breeding of animals is carried out.
  • utilising such a technique in artificial breeding would offer considerable economic advantages in livestock management, but is currently made difficult by the asymmetric geometry of the flat sperm head.
  • bull sperm are flat cells with head dimensions of approximately 10 microns by 4 microns by 1 micron attached to a 40 micron flagellum.
  • the asymmetric properties of the bull sperm head result in a high variation in both scattered light and fluorescent light emission with sperm orientation.
  • fluorescent emission varies by a factor of two with sperm orientation (see DNA Contention Measurements of Mammalian Sperm. CYTOMETRY 3:1-9 [1982]), effectively masking the 4% variation in intensity due to the sex of the sperm.
  • a number of flow cytometric systems have been developed in an attempt to overcome the problems encountered when analysing asymmetric particles such as sperm cells.
  • One flow cytometric system that has been developed in an attempt to overcome this problem introduces asymmetric cells travelling in a slow moving stream into the middle of a fast flowing sheath stream. Hydrodynamics then tends to align the asymmetric cells with their long axis parallel to the direction of the fast flowing sheath stream.
  • lasers typically deliver a single wavelength of light
  • use of lasers also has made it difficult to utilise a single light source to provide a variety of wavelengths of light, e.g. in conjunction with filters that filter out all but the desired wavelengths.
  • an optical apparatus including: a prism having a conical portion with an apex at a forward end of the prism and a central axis extending through the apex of the prism; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct an incident beam of electromagnetic radiation onto the apex of the conical portion in a direction substantially aligned with the central axis of the conical portion; and a reflective surface provided behind the apex of the prism; such that the beam refracted by the prism will be reflected by the reflective surface back through the prism to project from the forward end of the prism as an annular beam of electromagnetic radiation.
  • the optical apparatus described above thereby serves to produce an annular beam of electromagnetic radiation from a single beam of electromagnetic radiation incident onto the apex of the conical portion.
  • the arrangement is such to provide the beam with a constant cross section to produce a cylindrical tube of light.
  • the prism may also include a cylindrical base portion at a rear end thereof which has a circular cross section corresponding to the cross section of the base of the conical portion.
  • an optical apparatus including: a prism having a pyramidal portion with an even number of inclined faces meeting at an apex at a forward end of the prism and a central axis extending through the apex an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct an incident beam of electromagnetic radiation onto the apex of the pyramidal portion in a direction substantially aligned with the central axis of the pyramidal portion; and a reflective surface provided behind the apex of the prism; such that the beam refracted by the prism will be reflected by the reflective surface back through the prism to project from the forward end of the prism as a number of parallel beams.
  • the pyramidal portion has an even number of inclined faces since the optical geometry is such that the beams cross the prism to reflect from the opposing face.
  • the number of the inclined faces is not limited. For example, there may be 4, 6, 8 ... 12 inclined triangular faces converging towards the apex of the pyramidal portion.
  • the pyramidal portion also includes a base portion with a cross section corresponding to the base of the pyramidal portion. For example, where the pyramid has four inclined faces an appropriate base portion would be a rectangular prism or a cube.
  • the reflective surface may be provided at the rear end of the prism.
  • the invention is not limited to this arrangement and may potentially be disposed within the prism itself.
  • Another preferred arrangement is for the reflective surface to be spaced from the base portion.
  • Another desirable feature is that this spacing be adjustable to provide a variable annular beam diameter.
  • the reflective surface is spaced from the prism the electromagnetic radiation may suffer losses from multiple interface reflection.
  • the transmission losses would be less than for a longer prism with the reflective surface provided at the rear end.
  • the prisms are manufactured from optical glass such as BK7 optical glass.
  • optical glass such as BK7 optical glass.
  • the application is intended for use with UV electromagnetic radiation, it is preferred to manufacture the prism from UV-suitable material such as fused silica.
  • the reflective surface be comprised of a UV-grade mirror to increase the transmission efficiency of the optical apparatus.
  • the optical apparatus may be used with ultra-violet radiation, preferably produced from a laser source.
  • the electromagnetic radiation may also include other wavelengths including those in the visible spectrum.
  • the incident electromagnetic radiation is in the form of a collimated beam.
  • the optical apparatus described above in connection with the first two aspects may desirably be used in combination with a paraboloidal reflector having an internal paraboloidal-shaped reflective surface and an optical axis.
  • a paraboloidal reflector having an internal paraboloidal-shaped reflective surface and an optical axis.
  • Such a reflector will be oriented to receive, on its reflective surface, the electromagnetic radiation projected from the forward end of the prism.
  • a paraboloidal- shaped reflective surface will have a focus at which all light parallel to the optical axis and incident onto the reflective surface will be directed.
  • the parallel electromagnetic radiation projected from the prism will be received onto the paraboloidal reflector to converge at the focus.
  • Such a concentration of electromagnetic radiation may have many useful and varied applications in the field of optics.
  • the invention is capable of providing radially symmetric illumination to the focus of the paraboloidal reflector.
  • radially symmetric means that for every beam of incident radiation to the focus, a substantially diametrically opposite beam will be incident to the focus.
  • Each beam of the radially symmetric illumination may have the same angle to the optical axis of the paraboloidal reflector.
  • a convergent disc of electromagnetic radiation onto the focus will be included in the definition of "radially symmetric".
  • Such a convergent disc can be achieved through the use of the first-described optical apparatus in combination with the paraboloidal reflector. Any object can be placed at the focus of the paraboloidal reflector for illumination and inspection.
  • the apparatus has particular application to flow cytometry in that a flow source may be provided to direct particles through the focus of the paraboloidal reflector.
  • the source of electromagnetic radiation may not be directed directly at the apex of the prism and the invention allows for the use of mirrors and other reflectors as desired.
  • a second reflector may be disposed between the prism and the paraboloidal reflector, the second reflector having reflective portions to reflect the incident beam from the source onto the apex of the prism and transmitting portions to transmit the beam(s) projected from the forward end of the prism.
  • the invention is not limited to the particular prisms described in the forgoing aspects of the invention.
  • Other optical configurations are envisaged to produce the projected annular beam or parallel beams of electromagnetic radiation.
  • other types of reflectors which focus incident radiation towards one or more foci could be adopted.
  • a third aspect of the invention provides an optical apparatus including: an optical configuration adapted to produce an annular beam of electromagnetic radiation having a central axis or plurality of beams of electromagnetic radiation wherein said plurality of beams are evenly spaced from a central axis; and a focussing reflector having an internal reflective surface having an optical axis and one or more foci, the reflector being oriented to receive, onto its reflective surface, the annular beam or the plurality of beams of electromagnetic radiation.
  • the optical element may comprise any known reflective axicons as well as the particular prisms described above which, in some cases are also axicons.
  • the axicon may comprise an inner conical mirror with forward reflective surfaces surrounded by an outer conical mirror with forward reflective surfaces wherein the optical axes of the two mirrors are aligned.
  • the reflective surfaces form the letter "W", hence the name w-axicon or waxicon.
  • the focussing reflector has an internal reflective surface which is paraboloidal in shape.
  • paraboloidal reflector used throughout the specification and the claims will be understood to mean “a reflector conforming to the shape of a paraboloid of revolution”. The term is also to be understood to mean “a portion of a full paraboloid of revolution”.
  • optical axis of a paraboloid such an axis may also be considered to be the parabolic or central axis of the paraboloid.
  • the apparatus may be incorporated into a flow cytometer including a flow source to produce a flow of particles to be analysed in which the flow source is adapted to direct the flow of particles substantially through one of the foci of the reflective surface.
  • the flow source can be adapted to substantially align the flow with the optical axis of the reflective surface.
  • an aperture may be provided in the focussing reflector for passage of the flow therebeyond.
  • the present invention will be used in a flow cytometer accommodating a sorting function.
  • the flow means may include a nozzle and the flow cytometer may incorporate electrostatic droplet deflection sorting apparatus below the aperture in the focussing reflector.
  • an optical method including: providing a prism having a conical portion with an apex at the forward end, a central axis extending through the apex and a reflective surface provided behind the apex of the prism; directing an incident beam of electromagnetic radiation onto the apex of the conical portion in a direction substantially aligned with the central axis of the conical portion to produce an annular beam of electromagnetic radiation projecting from the forward end of the prism.
  • an optical method including: providing a prism having a pyramidal portion with an even number of inclined faces meeting at an apex at a forward end of the prism, a central axis extending through the apex and a reflective surface provided behind the apex of the prism; directing an incident beam of electromagnetic radiation onto the apex of the pyramidal portion in a direction substantially aligned with the central axis of the pyramidal portion to produce parallel beams of electromagnetic radiation projecting from the forward end of the prism.
  • an analysation instrument including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to converge substantially coplanar, substantially radially symmetric electromagnetic radiation towards the inspection zone.
  • the electromagnetic radiation coverges in the form of a disc disposed symmetrically relative to the central axis.
  • a method of analysing including: providing a flow of particles to be analysed; directing the flow of particles to be analysed through an inspection zone; converging substantially coplanar, substantially radially symmetric electromagnetic radiation towards the inspection zone.
  • an analysation instrument including: a flow source to produce a flow of particles to be analysed; a source of electromagnetic radiation; a reflector adapted to reflect at least a portion of the electromagnetic radiation at the flow of particles to illuminate the flow of particles; an optical configuration including a sensor adapted to sense electromagnetic radiation; wherein the reflector is also adapted to reflect, to the optical configuration, any electromagnetic radiation produced as a result of the illumination of the flow of particles.
  • the reflector described in accordance with this aspect serves the dual purpose of reflecting the electromagnetic radiation onto the flow of particles as well as collecting the electromagnetic radiation for transmission to the sensor.
  • Such a configuration can be achieved with the use of a reflector having an internal reflective surface which is paraboloidal in shape.
  • any use of the term “illumination” or “illuminate” is not restricted to merely visible illumination as non-visible wavelengths may also be used. As mentioned previously, in certain applications ultra violet radiation may be used. Furthermore, reference to electromagnetic radiation "produced” by the particle may include any florescence produced by the particles as a result of the incident illumination and/or any light scattered by the particles. It should also be understood that “irradiate” is intended to have the same meaning as “illuminate”.
  • a method of analysing including providing: a flow of particles to be analysed; providing a source of electromagnetic radiation; reflecting with a reflector at least a portion of the electromagnetic radiation to illuminate the flow of particles; reflecting with the reflector at least a portion of any electromagnetic radiation produced from the illumination of the flow of particles; sensing a portion of the electromagnetic radiation produced from the illumination of the flow of particles.
  • a flow cytometer including: a flow source to produce a linear flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement adapted to converge electromagnetic radiation onto the flow at the inspection zone in a radially symmetric manner about the inspection zone; a collector to collect electromagnetic radiation either produced or deflected from the particles in the flow; a processor to derive, from the collected electromagnetic radiation, predetermined information relating to each of at least some of the particles in the flow; and a correlator to correlate the derived information with the associated particle downstream of the inspection zone.
  • the radially symmetric illumination may be provided in the form of a continuous disc convergent towards the inspection zone.
  • Another preferred radially symmetric arrangement of the illumination is in the form of discreet beams converging towards the inspection zone. Either way, the particle is illuminated evenly from all sides.
  • a flow cytometer including: a flow source to produce a linear flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; and an optical arrangement including a focussing reflector having an internal reflective surface with one or more foci, the optical arrangement adapted to converge electromagnetic radiation onto the flow of particles at the inspection zone by reflection from the focussing reflector, the focussing reflector being oriented such that one of the one or more foci is substantially coincident with or located within the inspection zone.
  • the focussing reflector comprises a paraboloidal reflector having an internal reflective surface of paraboloidal-shape.
  • the flow of particles will thus flow through the focus of the paraboloidal reflector at which the electromagnetic radiation is converged.
  • the focussing reflector may have an ellipsoidal reflective surface with two foci and an optical axis extending between the two foci.
  • the flow source is oriented so that the flow of particles is aligned with the optical axis of the reflective surface.
  • any forms of the focussing reflector may be provided with an aperture for the passage of flow beyond the focussing reflector.
  • Such an embodiment is particularly adapted for use in a sorting flow cytometer which collects the electromagnetic radiation produced from the particles in the flow, processes the collected electromagnetic radiation to derive predetermined information relating to each of at least some of the particles in the flow and correlates the derived information with the associated particle downstream of the inspection zone.
  • the sorting flow cytometer can not only analyse the particles in the flow but sort the particles according to predetermined sets of selection criteria.
  • a preferred type of sorting flow cytometer is a jet-in-air flow cytometer.
  • a flow cytometer including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles, at the inspection zone; a collector to collect electromagnetic radiation either produced or deflected from the particles, the collector having an internal reflective surface with an optical axis and one or more foci, wherein the collector is oriented such that the flow of particles is substantially aligned with the optical axis.
  • a flow cytometer including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles, at the inspection zone; a collector to collect electromagnetic radiation either produced or deflected from the particles, the collector having an internal reflective surface with an optical axis and one or more foci, wherein the collector is disposed such that one of the one or more foci is substantially coincident or located within the inspection zone; a processor to derive, from the collected electromagnetic radiation, predetermined information relating to each of at least some of the particles in the flow; and a correlator to correlate the derived information with the associated particle downstream of the inspection zone.
  • the collector may be of the same form as the focussing reflector as described in accordance with previous aspects of the invention.
  • the collector may also comprise part of the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles.
  • the collector may serve the dual function of collecting the produced electromagnetic radiation as well as reflecting the incident radiation onto the particles.
  • an analysation instrument including: a first reflector having a partial ellipsoidal shape; a near focal point of the partial ellipsoidal shape of the first reflector; a distant focal point of the partial ellipsoidal shape of the first reflector; a central axis of the partial ellipsoidal shape defined by the near focal point and distant focal point of the partial ellipsoidal shape of the first reflector; a source of electromagnetic radiation disposed at the near focal point of the partial ellipsoidal shape capable of emitting electromagnetic radiation toward the first reflector; a second reflector having a partial ellipsoidal shape oriented relative to the first reflector so as to be capable of receiving electromagnetic radiation reflected by the first reflector; a near focal point of the partial ellipsoidal shape of the second reflector; a distant focal point of the partial ellipsoidal shape of the second reflector; a central axis of the partial ellipsoidal shape defined by the near focal point and distant focal point of
  • the source of electromagnetic radiation may comprise an arc lamp.
  • a preferred relationship between the first reflector and the second reflector is that the distant focal point of the first reflector and the distant focal point of the second reflector overlap.
  • the focal lengths of the first and second reflectors may be equivalent.
  • the focal lengths of the two reflectors may be different in that the first reflector has a greater focal length than the second reflector.
  • ellipsoidal reflector as used in the above described aspect of the invention and in following aspects and in the following description of the invention, is understood to mean a reflector which conforms to the shape of an ellipsoid of revolution. Furthermore, the term is understood to mean a portion of a full ellipsoid of revolution such as one third of an ellipsoid of revolution with an opening at the vertex.
  • the near focal point is intended to mean the focal point closest to the ellipsoidal portion being used.
  • a method of analysing including: utilising a first reflector having a partial ellipsoidal surface with a near focal point and a distant focal point; emitting electromagnetic radiation from a source of electromagnetic radiation positioned at the near focal point of the first reflector; reflecting electromagnetic radiation emitted by the source of electromagnetic radiation from the first reflector; utilising a second reflector having a partial ellipsoidal surface with a near focal point and a distant focal point; providing a flow of particles to be analysed; directing the flow of particles through an inspection zone; positioning the second reflector so that the near focal point of the second reflector overlaps the inspection zone and so that the second reflector is capable of receiving electromagnetic radiation reflected by the first reflector.
  • an analysation instrument including: a first reflector having a partial paraboloidal shape; a focal point, and a focal length of the partial paraboloidal shape of the first reflector; a parabolic axis of the partial paraboloidal shape of the first reflector; a source of electromagnetic radiation disposed at the focal point of the partial paraboloidal shape adapted to emit electromagnetic radiation toward the first reflector; a second reflector having a partial paraboloidal shape oriented relative to the first reflector so as to be capable of receiving electromagnetic radiation reflected by the first reflector; a focal point, and a focal length of the partial paraboloidal shape of the second reflector; a parabolic axis of the partial paraboloidal shape of the second reflector; a flow source to produce a flow of particles to be analysed; and an inspection zone of the flow of particles located at the focal point of the partial paraboloidal shape of the second reflector.
  • An arc lamp may be the source of electromagnetic radiation. It is preferred that the parabolic axes, i.e., optical axes, of the first and second paraboloidal-shapes are colinear. In one embodiment of the invention the focal lengths of the first and second reflectors may be equivalent. Alternatively the focal length of the first reflector may be greater than the focal length of the second reflector. A filter may be arranged between the focal points of the two reflectors.
  • a method of analysing including: utilising a first reflector having a partial paraboloidal surface, an optical axis and a focal point; emitting electromagnetic radiation from a source of electromagnetic radiation positioned at the focal point of the first reflector; reflecting electromagnetic radiation emitted by the source of electromagnetic radiation from the first reflector; utilising a second reflector having a partial paraboloidal surface, an optical axis and a focal point; providing a flow of particles to be analysed; directing the flow of particles through an inspection zone; positioning the second reflector so that the focal point of the second reflector overlaps the inspection zone and so that the second reflector is capable of receiving electromagnetic radiation reflected by the first reflector.
  • the present invention also provides, in accordance with another aspect of the invention, a nozzle including an opening for a flow of particles to flow through; a reflector coupled to the nozzle and oriented to reflect electromagnetic radiation at the flow of particles.
  • the reflector may take on various forms such as an ellipsoidal reflective surface or a paraboloidal reflective surface, the reflector and the nozzle may even be integral.
  • the flow of particles passes through an inspection zone and a source of electromagnetic radiation is provided to illuminate the inspection zone.
  • the reflective surface is of the kind having a focal point, then it is preferred that the focal point of the reflective surface overlaps the inspection zone.
  • the reflective surface may comprise a metal shape embedded in the nozzle.
  • the reflective surface may comprise a reflective coating applied to the nozzle.
  • the focal point of the reflective surface can be external to the nozzle.
  • the nozzle may be adapted to receive electromagnetic radiation through the opening in the nozzle to illuminate the reflector or through the nozzle material itself, e.g. via light transmission through a glass nozzle.
  • a method of illuminating a flow of particles including: providing a nozzle having a reflector coupled to the nozzle and oriented to reflect electromagnetic radiation; supplying a flow of particles; directing the flow of particles through the nozzle; reflecting electromagnetic radiation with the reflector toward the flow of particles.
  • a flow cytometer including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles, at the inspection zone; a partial ellipsoidal collector to collect electromagnetic radiation either produced or deflected from the particles, the collector having an internal reflective surface of partial ellipsoidal shape with two foci and an optical axis oriented along a line between the two foci; the flow source being oriented such that the flow of particles is substantially aligned with the optical axis.
  • the preferred form of the flow cytometer may be a jet-in-air flow cytometer. Most preferably, the flow cytometer enables sorting through the use of electrostatic plates.
  • a corresponding aspect of the invention provides a method of flow cytometry including passing a flow of particles to be analysed through an inspection zone; providing a focussing reflector having one or more foci; converging electromagnetic radiation onto the flow of particles at the inspection zone by reflection from the focussing reflector and aligning the inspection zone with one of the one or more foci.
  • Figure 1 (a) is a cross-sectional view of one embodiment of an optical apparatus capable of producing an annular beam of electromagnetic radiation
  • Figure 1(b) is a section through the beam of Figure 1 ;
  • Figure 1 (d) is a perspective view of one embodiment of a prism for use in the optical apparatus of Figure 1(a) ;
  • Figure 1(e) is a perspective view of an alternative form of a prism for use in the optical apparatus of figure 1(a);
  • Figure 1(f) is a perspective view of an alternative prism arrangement for use in the optical apparatus of Figure 1(a);
  • Figure 1 (g) is a perspective view of an alternative prism arrangement for use in the optical apparatus of Figure 1(a);
  • Figure 2 is sectional view of a paraboloidal reflector
  • Figure 3 shows various views though a reflector which includes transmitting and reflecting surfaces
  • Figure 4 is a cross-sectional view of a possible embodiment for a reflector apparatus
  • Figure 5 is a cross-sectional view of a possible embodiment for a detector apparatus
  • Figure 6 is a cross-sectional view of one preferred embodiment of a flow cytometer in accordance with an aspect of the present invention.
  • Figure 7 is a cross-sectional view of a second embodiment of a flow cytometer in accordance with an aspect of the present invention.
  • Figure 8 is a cross-sectional view of a third embodiment of a flow cytometer in accordance with an aspect of the present invention.
  • Figure 9 is a cross-sectional view of a fourth embodiment of a flow cytometer in accordance with an aspect of the present invention.
  • Figure 10 is a cross-sectional view of a fifth embodiment of a flow cytometer in accordance with an aspect of the present invention.
  • Figure 11 is a cross-sectional view of a sixth embodiment of a flow cytometer in accordance with an aspect of the present invention.
  • Figure 12 is a cross-sectional view of a reflector incorporated into a flow nozzle design according to an aspect of the present invention
  • Figure 13 is a cross-sectional view of a seventh embodiment of a flow cytometer in accordance with an aspect of the present invention
  • Figure 14 is a cross-sectional view of an eighth embodiment of a flow cytometer in accordance with an aspect of the present invention.
  • Figure 15 is a cross-sectional view of a ninth embodiment of a flow cytometer in accordance with an aspect of the present invention.
  • Figure 1(a) illustrates an optical apparatus including a prism 1.
  • the prism 1 has an apex 2 at a forward end of the prism, a right conical portion having a conical face 2, and a right cylindrical base portion contiguous with the conical portion.
  • the base portion has a circular shaped rear end 4 with a reflective coating.
  • An optical arrangement is provided to provide incoming electromagnetic radiation 5 such as ultra-violet light from a laser light source.
  • the UV light 5 is directed in direction aligned with the central axis of the prism 1 onto the apex 2 of the prism 1 via a second reflector in the form of mirror 6 positioned at an angle of 45 degrees with respect to the incoming light 5 and the central axis of the prism 1.
  • the prism 22 has a right pyramidal portion with four inclined faces meeting at an apex.
  • a base portion is also provided which is square in cross-section, corresponding to the cross-section of the base of the pyramidal portion.
  • the prism can be used in the same manner as prism 1 by directing incident light onto the apex of the prism in line with the central axis of the prism. However, in this embodiment, the projected light will emerge as four parallel beams equally spaced from the central axis.
  • the number of inclined faces of the pyramidal portion may vary, provided that an even number is maintained.
  • Figure 1(f) illustrates an alternative prism arrangement in which a reflective surface may be spaced from the rear end of the conical prism shown in Figure 1(e) or the pyramidal prism shown in Figure 1(d).
  • the spacing of the reflective surface 27 from the prism may be adjustable.
  • Figure 1(g) illustrates an alternative prism arrangement known as a w-axicon or waxicon.
  • the waxicon 28 comprises an inner conical axicon surrounded by an annular axicon concentric with the inner axicon.
  • the reflective surfaces define a W, hence the name waxicon.
  • Figure 2 shows a paraboloidal reflector 20 in the form of a mirror having a paraboloidal-shaped internal reflective surface.
  • the paraboloidal internal reflective surface has a focus and an optical axis running through the focus.
  • the paraboloidal shaped reflective surface can have the property whereby any light which leaves the focus of the paraboloidal reflector and becomes incident on the surface of the reflector will be reflected out of the reflector 20 parallel to the optical axis.
  • An aperture 21 is centrally positioned within the paraboloidal reflector 20, in line with the optical axis.
  • the paraboloidal reflector 20 may be used to provide multi-directional illumination of an object for analysis or inspection.
  • the incident light can be reflected towards the object at the focus. Further, if the incoming parallel light is evenly spaced in relation to the optical axis then the light illuminating the object at the focus will be radially symmetric.
  • the paraboloidal reflector 20 may thus be teamed with the optical apparatus shown in Figure 1 in a manner in which the paraboloidal reflector 20 is oriented to receive the light projected from the forward end of the prism 1 with the central axis of the prism 1 aligned with the optical axis of the paraboloidal reflector 20.
  • This particular arrangement is discussed further in connection with the flow cytometer shown in Figures 6, 7,9,10,11,13.
  • the paraboloidal reflector is not limited in its use in combination with the optical apparatus shown in Figure 1.
  • FIGs 3(a) (i) and (ii) are plan views of another embodiment of the second reflector of Figure 1 generally indicated by arrow 30.
  • the mirror 30 includes reflective surfaces 31 and 32.
  • the mirror 30 also includes a transmitting portion which is in the form of an annular ring 33. It should be appreciated that in some embodiments the transmitting portion 33 may be in the form of an aperture which extends through the mirror 30. However, in other embodiments such as that shown more clearly in Figure 3(b), the transmitting portion 33 may be in the form of a transparent material, such as glass 34 which has not been covered by a reflective surface 35.
  • any incoming light 36 that impacts on the reflective surface 35 is reflected, whereas incoming light which impacts on the transmitting portion 33 may continue to travel substantially in the same direction
  • the transmitting portion 33 when arranged at a 45 degree angle from which it is viewed in plan in Figure 3(a) (i) serves to allow passage of the annular beam of light projected from the forward end of the prism.
  • Figure 3(a) (ii) shows a plan view of the second reflector having an egg-shaped transmitting portion 33 necessary to achieve the annular transmitting portion 33 when oriented at 45 degrees.
  • Figure 4 shows an alternative reflector apparatus generally indicated by arrow 40 which may be used to collect illumination reflected from the paraboloidal reflector 20 in Figure 2.
  • the reflector apparatus 40 includes a body 46 having a number of reflective surfaces 41, 42 and 43 which are positioned with respect to the detector apparatus 40 so that they may reflect any light they receive in different directions and/or at different angles.
  • the reflector apparatus 40 also includes within its body 46 regions 44 and 45 (both of which are bounded by dotted lines) which allow for the transmission of light 47 through the reflector apparatus 40.
  • the regions 44 and 45 may be in the form of apertures through the body 46 or alternatively made of a transparent substance/material capable of allowing for the transmission of light.
  • regions 44 and 45 are made of a transparent substance/material it will usually be desirable that the regions have the same length as shown by double headed arrow x to ensure distance travelled and refraction of the light 47 is substantially identical in both regions.
  • the reflective surfaces 41, 42, and 43 are capable of discriminating against the different types of light A, B and C that may be received by the reflector apparatus 40, by reflecting it in different directions and/or at different angles.
  • the different types of light A, B and C may be reflected to suitable light detectors (not shown) for determination of the characteristics of each type of light.
  • Figure 5 illustrates a detector apparatus generally indicated by arrow 50 which may also be used to collect illumination from the paraboloidal reflector shown in Figure 2.
  • the detector apparatus 50 may also provide for the transmission of light 51 from a light source (not shown) in a similar manner to the reflector apparatus described above in connection with Figures 3 and 4.
  • the detector apparatus 50 may also have a number of light detectors 52, 53 and 54 spatially positioned so that they may receive the different types of light A, B and C incident on the reflector apparatus 50.
  • the spatial orientation of the light detectors 52, 53 and 54 on the detector apparatus 50 allows for the discrimination between different types of light.
  • measurement of certain light is not desired, eg. light merely reflected from the light source, such light can be allowed to travel through the transmitting portion(s) 51 of the detector apparatus.
  • FIG 6 illustrates a first preferred embodiment of a flow cytometer generally indicated by arrow 70.
  • the flow cytometer 70 includes the optical apparatus substantially as shown in Figure 1.
  • the optical apparatus includes an optical arrangement including a light source 71 and a mirror 72.
  • the light source 71 produces collimated ultra-violet laser light 73 which is directed via mirror 72 to a prism 74 having a central axis.
  • the prism 74 is configured to produce a cylinder of light 75 having a longitudinal axis coincident with the central axis of the prism.
  • the prism may be the same as that indicated in Figure 1(a) or (e) of the drawings.
  • the prism may have a pyramidal face such as that shown in Figure 1 (d) to produce parallel beams of light evenly spaced from the central axis of the prism.
  • the projected light 75 passes through an annular gap 76 in a second reflector 77 so as to be incident on the 45 degree point of a paraboloidal reflector/collector 78.
  • the reflector also services as a collector.
  • the paraboloidal reflector/collector 78 will be simply referred to as the paraboloidal reflector 78.
  • the paraboloidal reflector 78 has an optical axis aligned with the central axis of the prism and a focus F lying on the optical axis.
  • a nozzle assembly 79 which delivers a particle stream 80 eg sperm cells, which is substantially aligned with the optical axis of the paraboloidal reflector and passes through an inspection zone located at the focus F.
  • the nozzle assembly 79 delivers the sperm cells in a saline sheath solution and may utilise any of the known jet-in-air techniques to produce a laminar-flow particle stream with the sperm flowing single file through the inspection zone at F.
  • the paraboloidal reflector 78 is designed with two criteria in mind. Firstly, the reflector 78 should be able to withstand the corrosive environment introduced by the saline sheath environment. Secondly, the reflector should be designed to maximise reflectance of light of the UV frequency. Either of a rhodium reflective coating or an AlSi0 2 reflective coating on a nickel substrate were found to be effective.
  • the effect of the cylinder of light 75 being incident at the 45 degree point of the paraboloidal mirror 78 is that it is reflected at 90 degrees so as to form a substantially coplanar disc of light which is convergent on the focal point F of the paraboloidal reflector.
  • this disc of light is able to interact with the particle stream 80 and illuminate the particles within the stream with substantially radially symmetric illumination.
  • the particles will fluoresce when illuminated.
  • stains is an accepted technique in sperm sexing since the number of molecules of stain bound will be equivalent to the number of molecules of DNA. This difference in uptake will yield a difference in the number of cells available for excitation and fluorescence. The difference in DNA content between X and Y sperm will yield a corresponding measurable difference in fluorescent light. Any of the known stains currently used for sperm sexing may be used. In particular, Hoechst 33342 which is of the bis-benzimidazole family shown below has been shown to provide the necessary X-Y differential resolution.
  • the second reflector 77 is positioned at a substantially 45 degree angle so as to reflect the scattered and/or fluoresced light towards a light detector in the form of a photo- multiplier tube 82.
  • the second reflector 77 as appropriate may comprise the forms illustrated in figures 3-5.
  • the fluorescent light is of interest and the light merely scattered from the sperm in the sample stream may be of little or no interest.
  • the fluorescent light will be of a different frequency and the separation of the two frequencies can be achieved through the use of a high pass filter 200 positioned before the photo-multiplier tube 82.
  • the separation of frequencies may be achieved through the use of a dichroic mirror to reflect only those frequencies of interest.
  • the dichroic mirror may be incorporated into the second reflector 77.
  • no filter is necessary.
  • an array of measurement detectors may be provided with an appropriate array of filters for measuring different forms of light.
  • the use of a second reflector in the form as that shown in Figure 4 allows for the separation of light from different parts of the paraboloidal reflector, it being possible to apply different filters to each of the separate light parts.
  • Light which has not interacted with the particles may be refracted by the medium which makes up the sample stream 80 and radiate as a disc in the opposite direction to the incoming light.
  • the particle stream will generally have a small diameter the resulting refraction of light by the medium will not be great.
  • this light will substantially retrace the path of the illuminating cylinder of light and exit through the annular gap 76 in the second reflector 77. This creates a simple yet effective beam dump.
  • the amount of light measured by the photo-multiplier tube is passed to a processor, e.g., a computer (not shown) to derive predetermined information such as an association between the amount of measured light and a property of the cell from each of at least some of the particles in the flow.
  • a processor e.g., a computer (not shown) to derive predetermined information such as an association between the amount of measured light and a property of the cell from each of at least some of the particles in the flow.
  • This information is then correlated by a correlator, such as a computer, with the corresponding particle downstream of the inspection zone to enable sorting of the particle depending whether it meets certain selection criteria. For example, male and female sperm may be sorted by sex.
  • the flow sorting technique uses electrostatics to charge and deflect a cell containing droplet as it passes through an electric field.
  • the droplet is created by a mechanical oscillation applied through a piezo-electric transducer thus perturbing the sample stream as it exits the nozzle 79.
  • Each individual droplet can be charged depending on the characteristics of its contained particle just prior to break-off by application of a voltage to the carrier fluid. Depending on its charge, the droplet will be deflected from its normal gravitational trajectory by oppositely charged plates 83.
  • To incorporate droplet sorting it may be necessary to provide a means by which to view the stream so as to count the number of droplet spacings between the inspection point (ie. the focal point F) and the break-off point of the droplets. This can usually be achieved by inserting a small periscope through the aperture 84 in the base of the paraboloidal reflector 78. Droplets which are not electrostatically deflected from the central path are collected directly below and flushed to waste.
  • FIG 7 there is provided an alternative flow cytometer generally indicated by arrow 100, this flow cytometer being substantially similar to the flow cytometer 70 shown in Figure 6. Therefore, for ease of reference, similar numbering has been used to illustrate the components used in this embodiment.
  • light measurement detectors may be suitable positioned so that they only receive this light.
  • the radial optics deliver a convergent disk of light at the excitation wavelength to the inspection point. Adjusting the vertical dimension of the radial focus is relatively simple if a concave or convex element is positioned in the laser beam in front of the axicon. However, broadening the focus laterally, while retaining sufficient light intensity at the focus for stain excitation and fluorescence, is not trivial.
  • the radial focus requires that the illumination light cylinder be altered to cause divergence tangentially around its circular cross-section. This would result in a lateral displacement of the incoming light disk thereby broadening the intensity distribution of the focal area.
  • Some optical elements were proposed to perform this function.
  • the first optical element would take the form of a radially etched diffraction grating. Such a component would successfully achieve the goal of lateral displacement with a minimal dispersive effect in the vertical profile of the focus.
  • the second optical element is a light shaping diffuser element. Implementation of this element into the radial optics design would result in both vertical and lateral focus broadening.
  • Other options include a diffractor or a cylindrical lens causing the beam to diffract sideways and broaden the focus.
  • Another approach is to use the focussing characteristics of the laser beam which is a Caussian beam where the depth of focus 1 is proportional to the focal length f and inversely proportional to the beam diameter D.
  • the variable L is defined as the half- height width of the flex density profile as plotted along the optical axis.
  • paraboloidal and ellipsoidal configurations of reflectors can be used to provide illumination of an inspection zone of a linear flow of particles.
  • One distinct advantage of this type of system is the ability to use a low cost arc lamp to replace the more expensive lasers commonly used in instruments of this type. Lasers are preferred in some devices because of the intensity of light that they can deliver. However, they have the disadvantage of only providing a specific wavelength of electromagnetic radiation. Arc lamps, however, are less expensive and can provide many different wavelengths of electromagnetic radiation in their emissions. Then, the proper wavelength can be selected by use of an inexpensive filter which filters out the undesired wavelengths of electromagnetic radiation.
  • FIG 8 shows an analysation instrument 201, such as a flow cytometer, in which a first reflector 200 having a partial ellipsoidal shape is disposed above a flow source which produces a flow 237 of particles to be analysed.
  • the reflector can be referred to as a partial ellipsoidal reflector as it is essentially a halved ellipsoid. Nevertheless, it is understood that given the contour of its surface it is recognized as ellipsoidal or similarly having a partial ellipsoidal shape.
  • This first reflector 200 has both a near focal point 202 disposed near the top of the ellipsoid shown in Figure 8 and a distant focal point 204 disposed below the partial ellipsoidal shape in Figure 8.
  • a central axis 208 of the partial ellipsoidal shape is defined by these two focal points.
  • a second reflector 216 can be disposed or oriented below the first reflector.
  • the second reflector can have a partial ellipsoidal shape.
  • the partial ellipsoidal shape can have a near focal point 220 disposed near the bottom of Figure 8 and a distant focal point 224 disposed overlapping or coincident with the distant focal point 204 of the first reflector.
  • the partial ellipsoidal shape of the second reflector can have a central axis 228 defined by its near and distant focal points.
  • the central axis 208 of the first reflector is substantially aligned with the central axis 228 of the second reflector.
  • a source of electromagnetic radiation such as an arc lamp 212 can be disposed at the near focal point of the first reflector 200. Due to the properties of an ellipsoid, electromagnetic radiation emitted by the source of electromagnetic radiation from the near focal point 202 and incident upon the first reflector 200 can be reflected back to the distant focal point of the first reflector.
  • the distant focal point 204 of the first reflector and the distant focal point 224 of the second reflector are coincident and the central axis 208 of the first reflector and the central axis 228 of the second reflector are collinear, this reflected light can continue on a path such that it is incident upon the second reflector 216.
  • the second reflector 216 can then reflect the light which travelled through the distant focal point 224 of the second reflector to the near focal point 220 of the second reflector. In this fashion a real image of the source of electromagnetic radiation located at the near focal point 212 of the first reflector is created at the near focal point 220 of the second reflector 216. Therefore, a very intense light source can be concentrated on the inspection zone 236 of the linear flow of particles when the inspection zone is located at the near focal point 220 of the second reflector. Furthermore, this allows an arc lamp to be used - as a source with collimated beams, such as a laser, is unnecessary due to the ability of the reflectors to create a real image of the source of the electromagnetic radiation. Plus, a filter, such as a dichroic filter 240, can be used to filter out any wavelengths of undesired electromagnetic radiation.
  • the fluorescence 215 can be reflected by the second reflector back towards a reflective surface, such as dichroic filter 240 which reflects the fluorescence to detector housing 244 to be detected. Because of the ellipsoidal geometry a converging set of beams is created - thus, there is no need for optics to focus the fluorescence on the detector.
  • Figure 8 also shows that a stream of cells can be deflected for sorting or analysation purposes as they fall through an opening in the second reflector 216.
  • the first reflector and second reflector are shown having focal lengths of fl and £2 respectively.
  • the real image of the arc lamp will be the same size as the actual arc lamp.
  • it is desirable to shrink the size of the real image of the arc source This is the case when there is a possibility of two cells being very close to one another in the inspection zone of the stream. In such a case, it can be important to reduce a real image so that incident radiation is incident upon only the cell under analysation and not a second cell nearby. This prevents fluorescence from a second cell which might give an incorrect analysis. There is more likelihood of cells being close by when the throughput of the analyser is increased.
  • the arrangement of Figure 8 could be used with only the bottom reflector and an alternative light source to illuminate the flow of particles. This might involve a laser directed at the flow of particles or off the reflective surface of the ellipsoidal reflector 216.
  • This is a unique arrangement in flow cytometry, because the flow of particles is aligned coaxially with the central axis of the ellipsoidal reflector 216 to pass through the near focal point of the ellipsoidal reflector 216. After the flow of particles passes through the focal point at which the particles are irradiated with electromagnetic radiation for the purpose of analysation, they can be sorted based upon their identifying characteristics.
  • Electrostatic plates can be provided and disposed below the opening in the ellipsoidal reflective surface to deflect the particles as they pass close to or between the electrostatic plates. This embodiment is particularly unique in jet-in-air types of flow cytometers.
  • a first reflector 200 having a partial paraboloidal shape, a focal point (or focus) 302 is disposed to reflect electromagnetic radiation from a source of electromagnetic radiation, such as arc lamp 312.
  • the source of electromagnetic radiation can be positioned at the focus of the paraboloid such that all emissions originating from the focus and incident on the partial paraboloid are reflected in collimated beams 313 toward a second reflector 316.
  • the first reflector 300 and the second reflector 316 each have parabolic axes 308 and 338 respectively.
  • a flow source 332 can provide a flow of particles 337 that flows through the focal point 320 of the second reflector 316.
  • the portion of the flow of particles that flows through the focal point can be referred to as the inspection zone 336 upon which the electromagnetic radiation is focused so as to analyse a cell falling through the inspection zone.
  • the stained cell When the incident electromagnetic radiation is incident upon a cell in the inspection zone, the stained cell can be caused to fluoresce. This fluorescence 315 can then be reflected by the second reflector 316 toward a reflector, such as dichroic mirror 340, which directs the fluorescence toward an optical apparatus 345 that focuses the fluorescence on a detector 344.
  • a reflector such as dichroic mirror 340
  • a nozzle 400 can be provided with a reflector coupled to the nozzle itself.
  • the reflector can even be integral to the nozzle. This presents a significant advantage to the user of the analysing apparatus as there is no need for alignment of the components since the coupling can accomplish that task.
  • a paraboloidal nozzle is shown.
  • the nozzle can be manufactured of a material such as glass that permits the transmission of electromagnetic radiation, such as visible light. Incident beams of electromagnetic radiation from a source of electromagnetic radiation, such as a laser source 520 in Figure 11 pass through the nozzle body and are incident on a reflector 402.
  • the reflector 402 is coupled to the nozzle itself rather than existing separate from the nozzle.
  • An opening 404 can be provided in the nozzle to allow a flow of particles 408 to flow through.
  • the reflector 402 can be oriented to reflect the incident electromagnetic radiation at the flow of particles 408.
  • Two possible shapes which can be used for the reflective surface of the reflector are a paraboloid and an ellipsoid.
  • a paraboloidal reflective surface 412 is shown while in Figure 11, an ellipsoidal reflective surface 512 is shown.
  • an inspection zone 416 can overlap a focal point(s) of the reflective surface, such as focal point 420 of the paraboloid of Figure 10 to produce the desired reflection patterns.
  • the nozzle can be used with a source of electromagnetic radiation, such as a laser source 520 as shown in Figure 11. However, it is also envisioned that an arc lamp or other source could be used as well.
  • the source of electromagnetic radiation emits beams 450 which can be directed at the reflective surface. When the electromagnetic radiation is incident upon a cell under analysis, fluorescence is created as shown by beams 451.
  • the nozzle body could be shaped in a paraboloidal or ellipsoidal shape and then coated with a reflective material 428 applied to the nozzle surface.
  • a reflector such as a metal reflector 424 could be inserted or embedded in the nozzle body as shown in Figure 12. It might even be possible to rely on refractive properties which cause internal reflection or even total internal reflection.
  • FIG 13 an embodiment is shown in which the nozzle is shaped such that the focal point 420 of the reflective surface is external to the nozzle. External is intended to means outside of or away from the nozzle border, In such an embodiment, electromagnetic radiation could be directed at the focal point without needing to traverse through the nozzle body.
  • the radial optics configuration for a flow cytometer 500 can combine 360 degree radial illumination and radially symmetric collection of fluorescence from particles or cells as they pass through the inspection point.
  • a glass cone 516 and a paraboloidal reflector 528 can be used.
  • the optical beam of a laser 508 can be steered onto the point of the glass cone. The beam can then be refracted into a divergent cone of light which is retro-reflected to produce a cylinder of laser light which encircles and is antiparallel to the input beam.
  • This light cylinder can then be reflected by a 45 degree elliptical ring mirror 512 and aligned parallel to the optical axis of the paraboloidal reflector 528.
  • the angle of incidence of the cylindrical beam at the reflector is 45 degrees, causing the beam to form a coplanar convergent disk perpendicular to and focused on the sample stream.
  • Stained cells can be carried by the sample stream through the radial excitation focus and caused to fluoresce.
  • Much of the fluorescence can be collected by the paraboloidal reflector and projected out in a collimated beam onto an aspheric condensing lens 504.
  • the lens can focus the fluorescent light to a spot which is imaged by a microscope objective 520 into a phomultiplier tube (PMT) 501 and filter housing.
  • PMT phomultiplier tube
  • Optical alignment of specimens flowing through the focal region of the paraboloidal reflector can be achieved by adjusting the flow cell position to maximise fluorescent signals from calibration microspheres.
  • the paraboloidal reflector can have a hole or opening in the base through which the sample stream can exit and where a jet observation camera and droplet sorting mechanism 532 can be situated.
  • FIG 15 a simplified version of the geometry of Figure 14 is shown.
  • the fluorescence collection elements can be retained to provide radially symmetric detection of cells as they pass through the inspection point of the flow cytometer.
  • Excitation of cells can be performed by steering a laser beam 608 onto the paraboloidal reflector 628 at an incidence angle that results in beam delivery from one direction similar to standard flow cytometer illumination. This can be accomplished by reflecting the beam off mirror 612.
  • Detection of cells can be performed by a paraboloidal reflector and aspheric lens combination.
  • a single PMT for example with a 400LP filter, can be positioned to collect all of the light focused by the aspheric lens.
  • the embodiment in Figure 15 is particularly useful as it does not require as extensive an alignment of optics as is required in other embodiments.
  • An ellipsoidal collector could also be used to deliver the laser light reflected from an adjusted mirror 612 and to reflect fluoresence to be collected at the PMT.
  • the embodiments in Figures 15 and 16 are particularly advantageous because of the simplistic substantially coaxial alignment of the reflector with the detector.

Abstract

Various optical apparatus provide a source of parallel light (7, 75). The parallel light (7, 75) is generally achieved by directing an incident beam at the apex of a prism (1, 22, 24, 26, 28). The prism may have varying configurations. One configuration has a forward conical face (24). Another configuration has a pyramidal forward end (22). Other configurations are also disclosed. The application also discloses the use of reflectors (20, 78, 216, 316, 400) having internal reflective surfaces shaped as three-dimensional figures of revolution, for example paraboloid or ellipsoid. The reflectors (20, 78, 216, 316) focus light incident onto the reflectors at one or more foci (F, 220, 320, 420). The reflectors may be used in combination with the optical apparatus including the prisms (1, 22, 24, 26, 28). The reflectors (20, 78, 216, 316) may be used in flow cytometers for focussing light at a sample stream (237, 337) passing through the focus (F, 220, 320, 420) of the reflector (20, 78, 216, 316). The collection of scattered and/or fluorescent light from an illuminated sample stream (237, 337) in a flow cytometer may be achieved with the use of a collector shaped as a figure of revolution e.g. paraboloid or ellipsoid. Various optical methods and methods for flow cytometry are also disclosed.

Description

OPTICAL APPARATUS
TECHNICAL FIELD
This invention relates to an optical apparatus. In particular, although not exclusively, this invention has application to the field of flow cytometry. However, it is to be understood that several of the inventive aspects have application beyond flow cytometry and may have broad application in the field of optics generally. For example, several aspects of the invention may be used in photometry or optical particle detection apparatus.
BACKGROUND ART
Generally when illuminating a particle or an object for analysis, the light source is directed onto the particle from a single direction. An analysis may be made of light reflected or produced by the particle eg. fluorescence to reveal certain properties of the particle. The particular portion of the particle illuminated depends on the orientation of the particle with respect to the light source. Where the particle or object is asymmetrical, the light measurements will vary depending on which portion is illuminated, making it difficult to analyse the particle or object as a whole.
Such difficulties are encountered in flow cytometry since it is common for particles being analysed to be asymmetrical eg. mammalian spermatozoa.
Flow cytometers are often used to measure the properties of cells or particles which are carried in a stream of fluid. The stream is generally comprised of a sheath fluid into the centre of which is injected a narrow aqueous suspension of cells/particles. The sheath fluid focuses the sample cells/particles into single file. The stream containing the particles/cells passes through an inspection point which is the focus of an intense light beam. The particles/cells may have been stained with a light - sensitive stain which when illuminated, will absorb the incident light and fluoresce. Light scatters off the particles and/or alternatively causes fluorescence. This scattered or fluorescent light is then measured by a detector generally aligned with the incident beam. The characteristics of the detected signal(s) such as peak intensity, peak area or other characteristics of interest may then be used to derive properties of the particle, for example size. In a flow cytometer with sorting capability (as opposed to a purely analytical instrument) the detected signal(s) may be used to trigger sorting hardware which can be programmed to divert droplets from the stream of fluid. The sorting criteria will vary with the application, for example, the sorting may be conducted according to size or, in the case of spermatozoa, the DNA content of the cell.
One problem with conventional flow cytometers is that particle asymmetry often renders the optical characteristics of a particle difficult to measure. For example, a flat particle can pass through the inspection point with a random orientation. Thus, the intensity of the resultant scattered or fluorescent light may vary according to particle orientation and the detectors will measure different light intensities at different locations.
Thus, particle asymmetry can lead to a reduced resolution of measurement of the particles. It follows that, in cytometers with a sorting capability, this reduced resolution in measurement of the particles results in a decreased ability to accurately separate populations of cells with different optical properties. Such a problem is encountered in separation of male and female mammalian sperm.
In mammals, sperm carry the sex determining chromosomes and the total DNA content found in male and female sperm may differ. For example, in cattle the difference in the DNA content between male and female sperm is approximately 4%. This difference in DNA provides a means by which sperm may be separated in a sorting flow cytometer, making a predetermination of an offspring's sex possible when artificial breeding of animals is carried out. Utilising such a technique in artificial breeding would offer considerable economic advantages in livestock management, but is currently made difficult by the asymmetric geometry of the flat sperm head. As an example, bull sperm are flat cells with head dimensions of approximately 10 microns by 4 microns by 1 micron attached to a 40 micron flagellum. The asymmetric properties of the bull sperm head result in a high variation in both scattered light and fluorescent light emission with sperm orientation. In particular, fluorescent emission varies by a factor of two with sperm orientation (see DNA Contention Measurements of Mammalian Sperm. CYTOMETRY 3:1-9 [1982]), effectively masking the 4% variation in intensity due to the sex of the sperm.
A number of flow cytometric systems have been developed in an attempt to overcome the problems encountered when analysing asymmetric particles such as sperm cells. One flow cytometric system that has been developed in an attempt to overcome this problem introduces asymmetric cells travelling in a slow moving stream into the middle of a fast flowing sheath stream. Hydrodynamics then tends to align the asymmetric cells with their long axis parallel to the direction of the fast flowing sheath stream.
While this approach tends to reduce the vertical variation of light intensity from asymmetric particles, the radial variation remains. This system has been further refined so as to further reduce the orientation-related variation in the detected light intensity of particles.
The system developed by Pinkel et al (see Flow Cytometry in Mammalian Sperm: Progress Morphology and DNA Measurement. THE JOURNAL OF HISTOCHEMISTRY AND CYTOCHEMISTRY 24:353-358 [1979]), showed that the orientation of bull sperm could be further aligned by bevelling the end of the tube which injected the sample stream (ie. that which contains the sperm) into the sheath flow.
The system which attempted to overcome the problems of flow cytometric analysis of asymmetric cells was that described by Johnson (see Sex Preselection by Flow Cytometric Separation of X AND Y Chromosome Bearing Sperm Based on DNA Difference: A review. REPRODUCTIVE FERTILITY DEVELOPMENTS 7:893-903 [1995]), in relation to separation of bull sperm by sex. Johnson's approach utilised two detectors; one in line with the illuminating laser beam (the 0 degree detector) and one at right angles to the beam (the 90 degree detector). Sperm emit fluorescence preferentially through their narrow edges. Johnson determined which sperm were aligned edge-on to the 90 degree detector by detecting the bright emission from their edges, and used the 0 degree detector for measuring the flat-face emission from only the aligned sperm.
However, this system still had a number of drawbacks. One drawback was that it was a requirement for this system that the sample flow be moving slowly with respect to the sheath flow, thereby reducing sample throughput. A further drawback was that it only produces good alignment at very low flow rates. At the optimal flow rate, which produced the greatest number of aligned cells per second, only 40% of cells were aligned. Thus, the number of aligned cells had been increased from 10% to 40%, but approximately 60% of the cells remained unaligned, and further, due to the requirement of a low flow rate, there was a reduction in system throughput. It will be appreciated that the rejection of unaligned cells again reduces the processing rate of this system and unnecessarily wastes sperm cells.
One system which moved towards radial light collection was the Ellipsoidal Collector described by Skogen-Hagenson et al (see A High Efficiency Flow Cytometer, CYTOCHEMISTRY 25:784-789 [1977]), who developed a light collection system based on a hollow "egg shaped" brass reflector. The reflector surface was elliptical in cross-section and light from the inspection point at one focus was collected at the second focus. This system was demonstrated to have an ability to reduce the orientation dependence observed with bull sperm.
However, it still had orientation dependent illumination, (ie. Light source coming from a single direction). A further problem with this system is that it is unable to provide a particle sort function (ie. according to sperm sex).
A further system which implemented both symmetric illumination and symmetric light collection was the Epi-Illumination system described by Garner et al (see Quantification of the X and Y Chromosome Bearing Spermatozoa of Domestic Animals by Flow Cytometry, BIOLOGY OF REPRODUCTION 28:312-321 [1983]). In this system the sample stream travelled directly towards a high numerical index microscope objective lens and was diverted sideways after the stream had passed through the focal point of the lens. Illumination was delivered through the lens and light was collected back through the lens.
While this system also demonstrated a good ability to eliminate the orientation dependencies of bull sperm, it was also incapable of modification for high speed sorting. This was due to its sideways diversion of the sperm immediately after passing through the focal point.
Earlier systems have also relied on laser light, because of the intensity of laser light sources. Unfortunately, such laser systems can be quite expensive and only add to the cost of devices such as flow cytometers. Because lasers typically deliver a single wavelength of light, use of lasers also has made it difficult to utilise a single light source to provide a variety of wavelengths of light, e.g. in conjunction with filters that filter out all but the desired wavelengths.
Furthermore, previous systems have often required the precise alignment of optics in order to accomplish a proper delivery of electromagnetic radiation onto the cell under analysation or collection of fluorescence emitted by a cell. This can be a tedious process that adds to the expense of the analysation instruments. Hence, there is a need for a system, e.g., in flow cytometry, in which the optics that focus and collect electromagnetic radiation for measurement purposes are quicly and easily established in their proper orientation.
It is an object of the present invention to overcome the afore mentioned shortcomings of known optical apparatus with particular application to flow cytometers. It is also an object of the invention to provide the public with a useful choice.
SUMMARY OF THE INVENTION
In accordance with a first aspect of the present invention there is provided an optical apparatus including: a prism having a conical portion with an apex at a forward end of the prism and a central axis extending through the apex of the prism; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct an incident beam of electromagnetic radiation onto the apex of the conical portion in a direction substantially aligned with the central axis of the conical portion; and a reflective surface provided behind the apex of the prism; such that the beam refracted by the prism will be reflected by the reflective surface back through the prism to project from the forward end of the prism as an annular beam of electromagnetic radiation.
The optical apparatus described above thereby serves to produce an annular beam of electromagnetic radiation from a single beam of electromagnetic radiation incident onto the apex of the conical portion. Preferably, the arrangement is such to provide the beam with a constant cross section to produce a cylindrical tube of light. The prism may also include a cylindrical base portion at a rear end thereof which has a circular cross section corresponding to the cross section of the base of the conical portion.
In accordance with a second aspect of the present invention there is provided an optical apparatus including: a prism having a pyramidal portion with an even number of inclined faces meeting at an apex at a forward end of the prism and a central axis extending through the apex an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct an incident beam of electromagnetic radiation onto the apex of the pyramidal portion in a direction substantially aligned with the central axis of the pyramidal portion; and a reflective surface provided behind the apex of the prism; such that the beam refracted by the prism will be reflected by the reflective surface back through the prism to project from the forward end of the prism as a number of parallel beams.
It is required that the pyramidal portion have an even number of inclined faces since the optical geometry is such that the beams cross the prism to reflect from the opposing face. Apart from this constraint , the number of the inclined faces is not limited. For example, there may be 4, 6, 8 ... 12 inclined triangular faces converging towards the apex of the pyramidal portion. Preferably, the pyramidal portion also includes a base portion with a cross section corresponding to the base of the pyramidal portion. For example, where the pyramid has four inclined faces an appropriate base portion would be a rectangular prism or a cube.
In either of the first two aspects of the invention, the reflective surface may be provided at the rear end of the prism. However, the invention is not limited to this arrangement and may potentially be disposed within the prism itself. Another preferred arrangement is for the reflective surface to be spaced from the base portion. Another desirable feature is that this spacing be adjustable to provide a variable annular beam diameter. However, where the reflective surface is spaced from the prism the electromagnetic radiation may suffer losses from multiple interface reflection. However, as such a design would have a reduced length from the front to the rear end, the transmission losses would be less than for a longer prism with the reflective surface provided at the rear end.
Suitably the prisms are manufactured from optical glass such as BK7 optical glass. However, where the application is intended for use with UV electromagnetic radiation, it is preferred to manufacture the prism from UV-suitable material such as fused silica. In such an application, it is also desirable that the reflective surface be comprised of a UV-grade mirror to increase the transmission efficiency of the optical apparatus.
As mentioned above, the optical apparatus may be used with ultra-violet radiation, preferably produced from a laser source. The electromagnetic radiation may also include other wavelengths including those in the visible spectrum. Suitably, the incident electromagnetic radiation is in the form of a collimated beam.
The optical apparatus described above in connection with the first two aspects may desirably be used in combination with a paraboloidal reflector having an internal paraboloidal-shaped reflective surface and an optical axis. Such a reflector will be oriented to receive, on its reflective surface, the electromagnetic radiation projected from the forward end of the prism. It will be appreciated that such a paraboloidal- shaped reflective surface will have a focus at which all light parallel to the optical axis and incident onto the reflective surface will be directed. In other words, the parallel electromagnetic radiation projected from the prism will be received onto the paraboloidal reflector to converge at the focus. Such a concentration of electromagnetic radiation may have many useful and varied applications in the field of optics. In particular, the invention is capable of providing radially symmetric illumination to the focus of the paraboloidal reflector. The term "radially symmetric" means that for every beam of incident radiation to the focus, a substantially diametrically opposite beam will be incident to the focus. Each beam of the radially symmetric illumination may have the same angle to the optical axis of the paraboloidal reflector. Thus a convergent disc of electromagnetic radiation onto the focus will be included in the definition of "radially symmetric". Such a convergent disc can be achieved through the use of the first-described optical apparatus in combination with the paraboloidal reflector. Any object can be placed at the focus of the paraboloidal reflector for illumination and inspection. As will be discussed with following aspects of the invention, the apparatus has particular application to flow cytometry in that a flow source may be provided to direct particles through the focus of the paraboloidal reflector.
It will be understood that the source of electromagnetic radiation may not be directed directly at the apex of the prism and the invention allows for the use of mirrors and other reflectors as desired. In particular, a second reflector may be disposed between the prism and the paraboloidal reflector, the second reflector having reflective portions to reflect the incident beam from the source onto the apex of the prism and transmitting portions to transmit the beam(s) projected from the forward end of the prism.
However, the invention is not limited to the particular prisms described in the forgoing aspects of the invention. Other optical configurations are envisaged to produce the projected annular beam or parallel beams of electromagnetic radiation. Furthermore, other types of reflectors which focus incident radiation towards one or more foci could be adopted.
Accordingly, a third aspect of the invention provides an optical apparatus including: an optical configuration adapted to produce an annular beam of electromagnetic radiation having a central axis or plurality of beams of electromagnetic radiation wherein said plurality of beams are evenly spaced from a central axis; and a focussing reflector having an internal reflective surface having an optical axis and one or more foci, the reflector being oriented to receive, onto its reflective surface, the annular beam or the plurality of beams of electromagnetic radiation.
For example, the optical element may comprise any known reflective axicons as well as the particular prisms described above which, in some cases are also axicons. For example, the axicon may comprise an inner conical mirror with forward reflective surfaces surrounded by an outer conical mirror with forward reflective surfaces wherein the optical axes of the two mirrors are aligned. The reflective surfaces form the letter "W", hence the name w-axicon or waxicon.
Preferably, the focussing reflector has an internal reflective surface which is paraboloidal in shape. The use of the term "paraboloidal reflector" used throughout the specification and the claims will be understood to mean "a reflector conforming to the shape of a paraboloid of revolution". The term is also to be understood to mean "a portion of a full paraboloid of revolution". Similarly, in regard to the optical axis of a paraboloid, such an axis may also be considered to be the parabolic or central axis of the paraboloid.
As mentioned in connection with the foregoing aspect of the invention, the apparatus may be incorporated into a flow cytometer including a flow source to produce a flow of particles to be analysed in which the flow source is adapted to direct the flow of particles substantially through one of the foci of the reflective surface. Suitably the flow source can be adapted to substantially align the flow with the optical axis of the reflective surface. Moreover, an aperture may be provided in the focussing reflector for passage of the flow therebeyond.
It is desirable that the present invention will be used in a flow cytometer accommodating a sorting function. Thus, the flow means may include a nozzle and the flow cytometer may incorporate electrostatic droplet deflection sorting apparatus below the aperture in the focussing reflector.
In accordance with a fourth aspect of the present invention there is provided an optical method including: providing a prism having a conical portion with an apex at the forward end, a central axis extending through the apex and a reflective surface provided behind the apex of the prism; directing an incident beam of electromagnetic radiation onto the apex of the conical portion in a direction substantially aligned with the central axis of the conical portion to produce an annular beam of electromagnetic radiation projecting from the forward end of the prism. In accordance with a fifth aspect of the present invention there is provided an optical method including: providing a prism having a pyramidal portion with an even number of inclined faces meeting at an apex at a forward end of the prism, a central axis extending through the apex and a reflective surface provided behind the apex of the prism; directing an incident beam of electromagnetic radiation onto the apex of the pyramidal portion in a direction substantially aligned with the central axis of the pyramidal portion to produce parallel beams of electromagnetic radiation projecting from the forward end of the prism.
In accordance with another aspect of the present invention there is provided an analysation instrument including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to converge substantially coplanar, substantially radially symmetric electromagnetic radiation towards the inspection zone.
Preferably, the electromagnetic radiation coverges in the form of a disc disposed symmetrically relative to the central axis.
In accordance with yet another aspect of the present invention there is provided a method of analysing including: providing a flow of particles to be analysed; directing the flow of particles to be analysed through an inspection zone; converging substantially coplanar, substantially radially symmetric electromagnetic radiation towards the inspection zone.
In accordance with a further aspect of the present invention there is provided an analysation instrument including: a flow source to produce a flow of particles to be analysed; a source of electromagnetic radiation; a reflector adapted to reflect at least a portion of the electromagnetic radiation at the flow of particles to illuminate the flow of particles; an optical configuration including a sensor adapted to sense electromagnetic radiation; wherein the reflector is also adapted to reflect, to the optical configuration, any electromagnetic radiation produced as a result of the illumination of the flow of particles.
Thus the reflector described in accordance with this aspect serves the dual purpose of reflecting the electromagnetic radiation onto the flow of particles as well as collecting the electromagnetic radiation for transmission to the sensor. Such a configuration can be achieved with the use of a reflector having an internal reflective surface which is paraboloidal in shape.
It will be understood that any use of the term "illumination" or "illuminate" is not restricted to merely visible illumination as non-visible wavelengths may also be used. As mentioned previously, in certain applications ultra violet radiation may be used. Furthermore, reference to electromagnetic radiation "produced" by the particle may include any florescence produced by the particles as a result of the incident illumination and/or any light scattered by the particles. It should also be understood that "irradiate" is intended to have the same meaning as "illuminate".
In accordance with a still further aspect of the present invention there is provided a method of analysing including providing: a flow of particles to be analysed; providing a source of electromagnetic radiation; reflecting with a reflector at least a portion of the electromagnetic radiation to illuminate the flow of particles; reflecting with the reflector at least a portion of any electromagnetic radiation produced from the illumination of the flow of particles; sensing a portion of the electromagnetic radiation produced from the illumination of the flow of particles.
In accordance with still a further aspect of the present invention there is provided a flow cytometer including: a flow source to produce a linear flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement adapted to converge electromagnetic radiation onto the flow at the inspection zone in a radially symmetric manner about the inspection zone; a collector to collect electromagnetic radiation either produced or deflected from the particles in the flow; a processor to derive, from the collected electromagnetic radiation, predetermined information relating to each of at least some of the particles in the flow; and a correlator to correlate the derived information with the associated particle downstream of the inspection zone.
As mentioned previously, the radially symmetric illumination may be provided in the form of a continuous disc convergent towards the inspection zone. Another preferred radially symmetric arrangement of the illumination is in the form of discreet beams converging towards the inspection zone. Either way, the particle is illuminated evenly from all sides.
In accordance with a further aspect of the present invention there is provided a flow cytometer including: a flow source to produce a linear flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; and an optical arrangement including a focussing reflector having an internal reflective surface with one or more foci, the optical arrangement adapted to converge electromagnetic radiation onto the flow of particles at the inspection zone by reflection from the focussing reflector, the focussing reflector being oriented such that one of the one or more foci is substantially coincident with or located within the inspection zone.
Various embodiments of the focussing reflector have been envisaged. In one such embodiment the focussing reflector comprises a paraboloidal reflector having an internal reflective surface of paraboloidal-shape. The flow of particles will thus flow through the focus of the paraboloidal reflector at which the electromagnetic radiation is converged. In another embodiment of the invention the focussing reflector may have an ellipsoidal reflective surface with two foci and an optical axis extending between the two foci. In particularly preferred versions of this, the flow source is oriented so that the flow of particles is aligned with the optical axis of the reflective surface. Moreover, any forms of the focussing reflector may be provided with an aperture for the passage of flow beyond the focussing reflector. Such an embodiment is particularly adapted for use in a sorting flow cytometer which collects the electromagnetic radiation produced from the particles in the flow, processes the collected electromagnetic radiation to derive predetermined information relating to each of at least some of the particles in the flow and correlates the derived information with the associated particle downstream of the inspection zone. In this way, the sorting flow cytometer can not only analyse the particles in the flow but sort the particles according to predetermined sets of selection criteria. A preferred type of sorting flow cytometer is a jet-in-air flow cytometer.
In another aspect of the present invention there is provided a flow cytometer including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles, at the inspection zone; a collector to collect electromagnetic radiation either produced or deflected from the particles, the collector having an internal reflective surface with an optical axis and one or more foci, wherein the collector is oriented such that the flow of particles is substantially aligned with the optical axis.
In yet another aspect of the present invention there is provided a flow cytometer including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles, at the inspection zone; a collector to collect electromagnetic radiation either produced or deflected from the particles, the collector having an internal reflective surface with an optical axis and one or more foci, wherein the collector is disposed such that one of the one or more foci is substantially coincident or located within the inspection zone; a processor to derive, from the collected electromagnetic radiation, predetermined information relating to each of at least some of the particles in the flow; and a correlator to correlate the derived information with the associated particle downstream of the inspection zone.
The collector may be of the same form as the focussing reflector as described in accordance with previous aspects of the invention. In fact, the collector may also comprise part of the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles. In other words the collector may serve the dual function of collecting the produced electromagnetic radiation as well as reflecting the incident radiation onto the particles.
In accordance with another aspect of the present invention there is provided an analysation instrument including: a first reflector having a partial ellipsoidal shape; a near focal point of the partial ellipsoidal shape of the first reflector; a distant focal point of the partial ellipsoidal shape of the first reflector; a central axis of the partial ellipsoidal shape defined by the near focal point and distant focal point of the partial ellipsoidal shape of the first reflector; a source of electromagnetic radiation disposed at the near focal point of the partial ellipsoidal shape capable of emitting electromagnetic radiation toward the first reflector; a second reflector having a partial ellipsoidal shape oriented relative to the first reflector so as to be capable of receiving electromagnetic radiation reflected by the first reflector; a near focal point of the partial ellipsoidal shape of the second reflector; a distant focal point of the partial ellipsoidal shape of the second reflector; a central axis of the partial ellipsoidal shape defined by the near focal point and distant focal point of the partial ellipsoidal shape of the second reflector; a flow source to produce a flow of particles to be analysed; and an inspection zone of the flow of particles located at the near focal point of the partial ellipsoidal shape of the second reflector.
In a preferred embodiment, the source of electromagnetic radiation may comprise an arc lamp. Further, a preferred relationship between the first reflector and the second reflector is that the distant focal point of the first reflector and the distant focal point of the second reflector overlap. The focal lengths of the first and second reflectors may be equivalent. Alternatively, the focal lengths of the two reflectors may be different in that the first reflector has a greater focal length than the second reflector.
The term "ellipsoidal reflector" as used in the above described aspect of the invention and in following aspects and in the following description of the invention, is understood to mean a reflector which conforms to the shape of an ellipsoid of revolution. Furthermore, the term is understood to mean a portion of a full ellipsoid of revolution such as one third of an ellipsoid of revolution with an opening at the vertex.
In referring to ellipsoids throughout this description where only a partial ellipsoid is used, the near focal point is intended to mean the focal point closest to the ellipsoidal portion being used.
In accordance with yet another aspect of the present invention there is provided a method of analysing including: utilising a first reflector having a partial ellipsoidal surface with a near focal point and a distant focal point; emitting electromagnetic radiation from a source of electromagnetic radiation positioned at the near focal point of the first reflector; reflecting electromagnetic radiation emitted by the source of electromagnetic radiation from the first reflector; utilising a second reflector having a partial ellipsoidal surface with a near focal point and a distant focal point; providing a flow of particles to be analysed; directing the flow of particles through an inspection zone; positioning the second reflector so that the near focal point of the second reflector overlaps the inspection zone and so that the second reflector is capable of receiving electromagnetic radiation reflected by the first reflector.
In accordance with another object of the present invention there is provided an analysation instrument including: a first reflector having a partial paraboloidal shape; a focal point, and a focal length of the partial paraboloidal shape of the first reflector; a parabolic axis of the partial paraboloidal shape of the first reflector; a source of electromagnetic radiation disposed at the focal point of the partial paraboloidal shape adapted to emit electromagnetic radiation toward the first reflector; a second reflector having a partial paraboloidal shape oriented relative to the first reflector so as to be capable of receiving electromagnetic radiation reflected by the first reflector; a focal point, and a focal length of the partial paraboloidal shape of the second reflector; a parabolic axis of the partial paraboloidal shape of the second reflector; a flow source to produce a flow of particles to be analysed; and an inspection zone of the flow of particles located at the focal point of the partial paraboloidal shape of the second reflector.
An arc lamp may be the source of electromagnetic radiation. It is preferred that the parabolic axes, i.e., optical axes, of the first and second paraboloidal-shapes are colinear. In one embodiment of the invention the focal lengths of the first and second reflectors may be equivalent. Alternatively the focal length of the first reflector may be greater than the focal length of the second reflector. A filter may be arranged between the focal points of the two reflectors.
In another aspect of the present invention there is provided a method of analysing including: utilising a first reflector having a partial paraboloidal surface, an optical axis and a focal point; emitting electromagnetic radiation from a source of electromagnetic radiation positioned at the focal point of the first reflector; reflecting electromagnetic radiation emitted by the source of electromagnetic radiation from the first reflector; utilising a second reflector having a partial paraboloidal surface, an optical axis and a focal point; providing a flow of particles to be analysed; directing the flow of particles through an inspection zone; positioning the second reflector so that the focal point of the second reflector overlaps the inspection zone and so that the second reflector is capable of receiving electromagnetic radiation reflected by the first reflector.
The present invention also provides, in accordance with another aspect of the invention, a nozzle including an opening for a flow of particles to flow through; a reflector coupled to the nozzle and oriented to reflect electromagnetic radiation at the flow of particles.
The reflector may take on various forms such as an ellipsoidal reflective surface or a paraboloidal reflective surface, the reflector and the nozzle may even be integral. In a preferred embodiment of the invention, the flow of particles passes through an inspection zone and a source of electromagnetic radiation is provided to illuminate the inspection zone. Where the reflective surface is of the kind having a focal point, then it is preferred that the focal point of the reflective surface overlaps the inspection zone.
In preferred forms of the invention, the reflective surface may comprise a metal shape embedded in the nozzle. Alternatively, the reflective surface may comprise a reflective coating applied to the nozzle. Suitably, the focal point of the reflective surface can be external to the nozzle. The nozzle may be adapted to receive electromagnetic radiation through the opening in the nozzle to illuminate the reflector or through the nozzle material itself, e.g. via light transmission through a glass nozzle. In accordance with a further aspect of the invention there is provided a method of illuminating a flow of particles, the method including: providing a nozzle having a reflector coupled to the nozzle and oriented to reflect electromagnetic radiation; supplying a flow of particles; directing the flow of particles through the nozzle; reflecting electromagnetic radiation with the reflector toward the flow of particles.
Another aspect of the invention provides a flow cytometer including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles, at the inspection zone; a partial ellipsoidal collector to collect electromagnetic radiation either produced or deflected from the particles, the collector having an internal reflective surface of partial ellipsoidal shape with two foci and an optical axis oriented along a line between the two foci; the flow source being oriented such that the flow of particles is substantially aligned with the optical axis.
The preferred form of the flow cytometer may be a jet-in-air flow cytometer. Most preferably, the flow cytometer enables sorting through the use of electrostatic plates.
A corresponding aspect of the invention provides a method of flow cytometry including passing a flow of particles to be analysed through an inspection zone; providing a focussing reflector having one or more foci; converging electromagnetic radiation onto the flow of particles at the inspection zone by reflection from the focussing reflector and aligning the inspection zone with one of the one or more foci.
BRIEF DESCRIPTION OF DRAWINGS
Further aspects of the present invention will become apparent from the following description which is given by way of example only and with reference to the accompanying drawings in which:
Figure 1 (a) is a cross-sectional view of one embodiment of an optical apparatus capable of producing an annular beam of electromagnetic radiation;
Figure 1(b) is a section through the beam of Figure 1 ;
Figure 1 (d) is a perspective view of one embodiment of a prism for use in the optical apparatus of Figure 1(a) ;
Figure 1(e) is a perspective view of an alternative form of a prism for use in the optical apparatus of figure 1(a); Figure 1(f) is a perspective view of an alternative prism arrangement for use in the optical apparatus of Figure 1(a); Figure 1 (g) is a perspective view of an alternative prism arrangement for use in the optical apparatus of Figure 1(a);
Figure 2 is sectional view of a paraboloidal reflector;
Figure 3 shows various views though a reflector which includes transmitting and reflecting surfaces;
Figure 4 is a cross-sectional view of a possible embodiment for a reflector apparatus;
Figure 5 is a cross-sectional view of a possible embodiment for a detector apparatus;
Figure 6 is a cross-sectional view of one preferred embodiment of a flow cytometer in accordance with an aspect of the present invention;
Figure 7 is a cross-sectional view of a second embodiment of a flow cytometer in accordance with an aspect of the present invention;
Figure 8 is a cross-sectional view of a third embodiment of a flow cytometer in accordance with an aspect of the present invention;
Figure 9 is a cross-sectional view of a fourth embodiment of a flow cytometer in accordance with an aspect of the present invention;
Figure 10 is a cross-sectional view of a fifth embodiment of a flow cytometer in accordance with an aspect of the present invention;
Figure 11 is a cross-sectional view of a sixth embodiment of a flow cytometer in accordance with an aspect of the present invention;
Figure 12 is a cross-sectional view of a reflector incorporated into a flow nozzle design according to an aspect of the present invention; Figure 13 is a cross-sectional view of a seventh embodiment of a flow cytometer in accordance with an aspect of the present invention;
Figure 14 is a cross-sectional view of an eighth embodiment of a flow cytometer in accordance with an aspect of the present invention; and
Figure 15 is a cross-sectional view of a ninth embodiment of a flow cytometer in accordance with an aspect of the present invention.
BESTMODES FOR CARRYING OUT THE INVENTION
Some embodiments of the invention are discussed in "A New Optical Configuration for Flow Cytometric Sorting of Aspherical Cells", Int. Soc. Optical Engr., Proc. Of Adv. Tech. Analytical Cytology, 1997, by John C. Sharpe, Peter N. Schaare and Rainer Kunnemeyer; "Radially Symmetric Excitation and Collection Optics for Flow Cytometric Sorting of Aspherical Cells", Cytometry 29:363-370 (1997) by John C. Sharpe, Peter N. Schaare, and Rainer Kunnemeyer; and "A New Optical Configuration for Flow Cytometric Sorting of Bovine Spermatozoa by Sex", a thesis submitted to the University of Waikato for the degree of Doctor of Philosophy in Physics by Johnathan Charles Sharpe, which are hereby incorporated by reference.
Figure 1(a) illustrates an optical apparatus including a prism 1. The prism 1 has an apex 2 at a forward end of the prism, a right conical portion having a conical face 2, and a right cylindrical base portion contiguous with the conical portion. The base portion has a circular shaped rear end 4 with a reflective coating. An optical arrangement is provided to provide incoming electromagnetic radiation 5 such as ultra-violet light from a laser light source. The UV light 5 is directed in direction aligned with the central axis of the prism 1 onto the apex 2 of the prism 1 via a second reflector in the form of mirror 6 positioned at an angle of 45 degrees with respect to the incoming light 5 and the central axis of the prism 1. As the incoming light 5 enters the prism 1 via the apex 2 it is refracted by the prism 1 and diverges in a cone and is reflected off the rear end 4 of the lens 1. The reflected light exits the prism 1 through its conical face 3 and is projected from the forward end of the prism as an annular beam. The beam defines an enclosed cylindrical band of light having a longitudinal axis coincident with the central axis of the prism 1. Figure 1(b) shows a cross section through the enclosed band of light. The production of a cylindrical band of light may have many uses throughout the field of optics. Figure 1(e) illustrates the prism 1 in perspective view. Figure 1 (d) illustrates an alternative form of prism 22. The prism 22 has a right pyramidal portion with four inclined faces meeting at an apex. A base portion is also provided which is square in cross-section, corresponding to the cross-section of the base of the pyramidal portion. The prism can be used in the same manner as prism 1 by directing incident light onto the apex of the prism in line with the central axis of the prism. However, in this embodiment, the projected light will emerge as four parallel beams equally spaced from the central axis. The number of inclined faces of the pyramidal portion may vary, provided that an even number is maintained.
Figure 1(f) illustrates an alternative prism arrangement in which a reflective surface may be spaced from the rear end of the conical prism shown in Figure 1(e) or the pyramidal prism shown in Figure 1(d). The spacing of the reflective surface 27 from the prism may be adjustable.
Figure 1(g) illustrates an alternative prism arrangement known as a w-axicon or waxicon. The waxicon 28 comprises an inner conical axicon surrounded by an annular axicon concentric with the inner axicon. The reflective surfaces define a W, hence the name waxicon.
Figure 2 shows a paraboloidal reflector 20 in the form of a mirror having a paraboloidal-shaped internal reflective surface. The paraboloidal internal reflective surface has a focus and an optical axis running through the focus. It will be understood that the paraboloidal shaped reflective surface can have the property whereby any light which leaves the focus of the paraboloidal reflector and becomes incident on the surface of the reflector will be reflected out of the reflector 20 parallel to the optical axis. Likewise, when light which is reflected parallel to the optical axis enters and hits the reflective surface, it will be projected toward and through the focus. An aperture 21 is centrally positioned within the paraboloidal reflector 20, in line with the optical axis.
Thus, the paraboloidal reflector 20 may be used to provide multi-directional illumination of an object for analysis or inspection. By positioning the object at the focus of the paraboloidal reflector 20 and providing light incident on the surface of the reflector 20 and parallel to the optical axis of the reflector 20, the incident light can be reflected towards the object at the focus. Further, if the incoming parallel light is evenly spaced in relation to the optical axis then the light illuminating the object at the focus will be radially symmetric. The paraboloidal reflector 20 may thus be teamed with the optical apparatus shown in Figure 1 in a manner in which the paraboloidal reflector 20 is oriented to receive the light projected from the forward end of the prism 1 with the central axis of the prism 1 aligned with the optical axis of the paraboloidal reflector 20. This particular arrangement is discussed further in connection with the flow cytometer shown in Figures 6, 7,9,10,11,13. However the paraboloidal reflector is not limited in its use in combination with the optical apparatus shown in Figure 1.
Figures 3(a) (i) and (ii) are plan views of another embodiment of the second reflector of Figure 1 generally indicated by arrow 30. The mirror 30 includes reflective surfaces 31 and 32. The mirror 30 also includes a transmitting portion which is in the form of an annular ring 33. It should be appreciated that in some embodiments the transmitting portion 33 may be in the form of an aperture which extends through the mirror 30. However, in other embodiments such as that shown more clearly in Figure 3(b), the transmitting portion 33 may be in the form of a transparent material, such as glass 34 which has not been covered by a reflective surface 35. As Figure 3(b) shows, any incoming light 36 that impacts on the reflective surface 35 is reflected, whereas incoming light which impacts on the transmitting portion 33 may continue to travel substantially in the same direction The transmitting portion 33 when arranged at a 45 degree angle from which it is viewed in plan in Figure 3(a) (i) serves to allow passage of the annular beam of light projected from the forward end of the prism. Figure 3(a) (ii) shows a plan view of the second reflector having an egg-shaped transmitting portion 33 necessary to achieve the annular transmitting portion 33 when oriented at 45 degrees.
Figure 4 shows an alternative reflector apparatus generally indicated by arrow 40 which may be used to collect illumination reflected from the paraboloidal reflector 20 in Figure 2. The reflector apparatus 40 includes a body 46 having a number of reflective surfaces 41, 42 and 43 which are positioned with respect to the detector apparatus 40 so that they may reflect any light they receive in different directions and/or at different angles.
The reflector apparatus 40 also includes within its body 46 regions 44 and 45 (both of which are bounded by dotted lines) which allow for the transmission of light 47 through the reflector apparatus 40. It should be appreciated that the regions 44 and 45 may be in the form of apertures through the body 46 or alternatively made of a transparent substance/material capable of allowing for the transmission of light. In embodiments where regions 44 and 45 are made of a transparent substance/material it will usually be desirable that the regions have the same length as shown by double headed arrow x to ensure distance travelled and refraction of the light 47 is substantially identical in both regions.
The reflective surfaces 41, 42, and 43 are capable of discriminating against the different types of light A, B and C that may be received by the reflector apparatus 40, by reflecting it in different directions and/or at different angles. Thus, the different types of light A, B and C may be reflected to suitable light detectors (not shown) for determination of the characteristics of each type of light.
Figure 5 illustrates a detector apparatus generally indicated by arrow 50 which may also be used to collect illumination from the paraboloidal reflector shown in Figure 2. In this embodiment the detector apparatus 50 may also provide for the transmission of light 51 from a light source (not shown) in a similar manner to the reflector apparatus described above in connection with Figures 3 and 4. The detector apparatus 50 may also have a number of light detectors 52, 53 and 54 spatially positioned so that they may receive the different types of light A, B and C incident on the reflector apparatus 50. Thus, the spatial orientation of the light detectors 52, 53 and 54 on the detector apparatus 50 allows for the discrimination between different types of light. On the other hand, where measurement of certain light is not desired, eg. light merely reflected from the light source, such light can be allowed to travel through the transmitting portion(s) 51 of the detector apparatus.
Figure 6 illustrates a first preferred embodiment of a flow cytometer generally indicated by arrow 70. The flow cytometer 70 includes the optical apparatus substantially as shown in Figure 1. The optical apparatus includes an optical arrangement including a light source 71 and a mirror 72. The light source 71 produces collimated ultra-violet laser light 73 which is directed via mirror 72 to a prism 74 having a central axis. The prism 74 is configured to produce a cylinder of light 75 having a longitudinal axis coincident with the central axis of the prism. The prism may be the same as that indicated in Figure 1(a) or (e) of the drawings. Alternatively, the prism may have a pyramidal face such as that shown in Figure 1 (d) to produce parallel beams of light evenly spaced from the central axis of the prism. The projected light 75 passes through an annular gap 76 in a second reflector 77 so as to be incident on the 45 degree point of a paraboloidal reflector/collector 78. It will be seen in the following discussion that the reflector also services as a collector. For ease of reference the paraboloidal reflector/collector 78 will be simply referred to as the paraboloidal reflector 78. The paraboloidal reflector 78 has an optical axis aligned with the central axis of the prism and a focus F lying on the optical axis. Situated within the paraboloidal reflector 78 is a nozzle assembly 79 which delivers a particle stream 80 eg sperm cells, which is substantially aligned with the optical axis of the paraboloidal reflector and passes through an inspection zone located at the focus F. The nozzle assembly 79 delivers the sperm cells in a saline sheath solution and may utilise any of the known jet-in-air techniques to produce a laminar-flow particle stream with the sperm flowing single file through the inspection zone at F.
The paraboloidal reflector 78 is designed with two criteria in mind. Firstly, the reflector 78 should be able to withstand the corrosive environment introduced by the saline sheath environment. Secondly, the reflector should be designed to maximise reflectance of light of the UV frequency. Either of a rhodium reflective coating or an AlSi02 reflective coating on a nickel substrate were found to be effective.
The effect of the cylinder of light 75 being incident at the 45 degree point of the paraboloidal mirror 78 is that it is reflected at 90 degrees so as to form a substantially coplanar disc of light which is convergent on the focal point F of the paraboloidal reflector. Thus, this disc of light is able to interact with the particle stream 80 and illuminate the particles within the stream with substantially radially symmetric illumination.
If the particles have been stained with light-sensitive stain, the particles will fluoresce when illuminated. The use of stains is an accepted technique in sperm sexing since the number of molecules of stain bound will be equivalent to the number of molecules of DNA. This difference in uptake will yield a difference in the number of cells available for excitation and fluorescence. The difference in DNA content between X and Y sperm will yield a corresponding measurable difference in fluorescent light. Any of the known stains currently used for sperm sexing may be used. In particular, Hoechst 33342 which is of the bis-benzimidazole family shown below has been shown to provide the necessary X-Y differential resolution.
Figure imgf000023_0001
Thus, light which interacts with the particles will be scattered and/or fluoresced. This scattered and/or fluoresced light is then collected by the paraboloidal reflector/collector 78 and reflected parallel to the optical axis of the paraboloidal reflector 78. The second reflector 77 is positioned at a substantially 45 degree angle so as to reflect the scattered and/or fluoresced light towards a light detector in the form of a photo- multiplier tube 82. The second reflector 77 as appropriate may comprise the forms illustrated in figures 3-5.
For the specific application of the present invention in sexing sperm, the fluorescent light is of interest and the light merely scattered from the sperm in the sample stream may be of little or no interest. The fluorescent light will be of a different frequency and the separation of the two frequencies can be achieved through the use of a high pass filter 200 positioned before the photo-multiplier tube 82. Alternatively, the separation of frequencies may be achieved through the use of a dichroic mirror to reflect only those frequencies of interest. For example the dichroic mirror may be incorporated into the second reflector 77. However, if in certain applications it is desirable to measure scattered light, no filter is necessary.
It should be appreciated that instead of the single measurement detector 82 shown, an array of measurement detectors may be provided with an appropriate array of filters for measuring different forms of light. For example, the use of a second reflector in the form as that shown in Figure 4 allows for the separation of light from different parts of the paraboloidal reflector, it being possible to apply different filters to each of the separate light parts.
Light which has not interacted with the particles may be refracted by the medium which makes up the sample stream 80 and radiate as a disc in the opposite direction to the incoming light. As the particle stream will generally have a small diameter the resulting refraction of light by the medium will not be great. Thus, this light will substantially retrace the path of the illuminating cylinder of light and exit through the annular gap 76 in the second reflector 77. This creates a simple yet effective beam dump.
It should be appreciated that supporting structures of the components of the flow cytometer 70 including sample flow tubes for the nozzle assembly may obscure parts of the path for the cylinder of light 75. However, any resulting asymmetry in the disc of light is generally negligible and the cylinder of light is therefore still considered cylindrical. Optics might even be provided to refract an incident beam around obstructions.
The amount of light measured by the photo-multiplier tube is passed to a processor, e.g., a computer (not shown) to derive predetermined information such as an association between the amount of measured light and a property of the cell from each of at least some of the particles in the flow. This information is then correlated by a correlator, such as a computer, with the corresponding particle downstream of the inspection zone to enable sorting of the particle depending whether it meets certain selection criteria. For example, male and female sperm may be sorted by sex.
The flow sorting technique uses electrostatics to charge and deflect a cell containing droplet as it passes through an electric field. The droplet is created by a mechanical oscillation applied through a piezo-electric transducer thus perturbing the sample stream as it exits the nozzle 79. Each individual droplet can be charged depending on the characteristics of its contained particle just prior to break-off by application of a voltage to the carrier fluid. Depending on its charge, the droplet will be deflected from its normal gravitational trajectory by oppositely charged plates 83. To incorporate droplet sorting it may be necessary to provide a means by which to view the stream so as to count the number of droplet spacings between the inspection point (ie. the focal point F) and the break-off point of the droplets. This can usually be achieved by inserting a small periscope through the aperture 84 in the base of the paraboloidal reflector 78. Droplets which are not electrostatically deflected from the central path are collected directly below and flushed to waste.
In Figure 7 there is provided an alternative flow cytometer generally indicated by arrow 100, this flow cytometer being substantially similar to the flow cytometer 70 shown in Figure 6. Therefore, for ease of reference, similar numbering has been used to illustrate the components used in this embodiment.
The major difference with this embodiment shown in Figure 7 is that only light 101 collected from the upper regions of the paraboloidal reflector are received by the photo- multipliers 102. Accordingly, the second reflector 77 need not be of the type discussed in the previous embodiment. Instead, only a continuous mirror confined within the cylindrical beam 75 need be used to reflect away the forward scattered and/or fluoresced light 103.
On the otherhand, it should also be readily appreciated that where it is only desirable to consider forward scattered and/or fluoresced light, light measurement detectors may be suitable positioned so that they only receive this light.
During experimentation, it was found that an increase in sample to sheath differential pressure resulted in increased positional uncertainty of the particles through the focus, which results in a difference in illumination, and therefore fluorescence emission. There are a number of possible solutions which may be used either singly or in combination to broaden the focus around the sample stream.
The radial optics deliver a convergent disk of light at the excitation wavelength to the inspection point. Adjusting the vertical dimension of the radial focus is relatively simple if a concave or convex element is positioned in the laser beam in front of the axicon. However, broadening the focus laterally, while retaining sufficient light intensity at the focus for stain excitation and fluorescence, is not trivial.
To laterally broaden or defocus the radial focus requires that the illumination light cylinder be altered to cause divergence tangentially around its circular cross-section. This would result in a lateral displacement of the incoming light disk thereby broadening the intensity distribution of the focal area. Some optical elements were proposed to perform this function. The first optical element would take the form of a radially etched diffraction grating. Such a component would successfully achieve the goal of lateral displacement with a minimal dispersive effect in the vertical profile of the focus. The second optical element is a light shaping diffuser element. Implementation of this element into the radial optics design would result in both vertical and lateral focus broadening. Other options include a diffractor or a cylindrical lens causing the beam to diffract sideways and broaden the focus.
Another approach is to use the focussing characteristics of the laser beam which is a Caussian beam where the depth of focus 1 is proportional to the focal length f and inversely proportional to the beam diameter D. The variable L is defined as the half- height width of the flex density profile as plotted along the optical axis. Thus, an increase in the focal length of the paraboloidal reflector will cause an increase in d. Also, decreasing the diameter of the illuminating laser beam will bring about an increase in d.
In another embodiment of the invention, paraboloidal and ellipsoidal configurations of reflectors can be used to provide illumination of an inspection zone of a linear flow of particles. One distinct advantage of this type of system is the ability to use a low cost arc lamp to replace the more expensive lasers commonly used in instruments of this type. Lasers are preferred in some devices because of the intensity of light that they can deliver. However, they have the disadvantage of only providing a specific wavelength of electromagnetic radiation. Arc lamps, however, are less expensive and can provide many different wavelengths of electromagnetic radiation in their emissions. Then, the proper wavelength can be selected by use of an inexpensive filter which filters out the undesired wavelengths of electromagnetic radiation.
Referring now to Figure 8, an ellipsoidal embodiment of the invention can be seen. Figure 8 shows an analysation instrument 201, such as a flow cytometer, in which a first reflector 200 having a partial ellipsoidal shape is disposed above a flow source which produces a flow 237 of particles to be analysed. The reflector can be referred to as a partial ellipsoidal reflector as it is essentially a halved ellipsoid. Nevertheless, it is understood that given the contour of its surface it is recognized as ellipsoidal or similarly having a partial ellipsoidal shape. This first reflector 200 has both a near focal point 202 disposed near the top of the ellipsoid shown in Figure 8 and a distant focal point 204 disposed below the partial ellipsoidal shape in Figure 8. A central axis 208 of the partial ellipsoidal shape is defined by these two focal points.
A second reflector 216 can be disposed or oriented below the first reflector. Again, the second reflector can have a partial ellipsoidal shape. Furthermore, the partial ellipsoidal shape can have a near focal point 220 disposed near the bottom of Figure 8 and a distant focal point 224 disposed overlapping or coincident with the distant focal point 204 of the first reflector. In addition, the partial ellipsoidal shape of the second reflector can have a central axis 228 defined by its near and distant focal points. Preferably, the central axis 208 of the first reflector is substantially aligned with the central axis 228 of the second reflector.
A source of electromagnetic radiation, such as an arc lamp 212 can be disposed at the near focal point of the first reflector 200. Due to the properties of an ellipsoid, electromagnetic radiation emitted by the source of electromagnetic radiation from the near focal point 202 and incident upon the first reflector 200 can be reflected back to the distant focal point of the first reflector. When the distant focal point 204 of the first reflector and the distant focal point 224 of the second reflector are coincident and the central axis 208 of the first reflector and the central axis 228 of the second reflector are collinear, this reflected light can continue on a path such that it is incident upon the second reflector 216. The second reflector 216 can then reflect the light which travelled through the distant focal point 224 of the second reflector to the near focal point 220 of the second reflector. In this fashion a real image of the source of electromagnetic radiation located at the near focal point 212 of the first reflector is created at the near focal point 220 of the second reflector 216. Therefore, a very intense light source can be concentrated on the inspection zone 236 of the linear flow of particles when the inspection zone is located at the near focal point 220 of the second reflector. Furthermore, this allows an arc lamp to be used - as a source with collimated beams, such as a laser, is unnecessary due to the ability of the reflectors to create a real image of the source of the electromagnetic radiation. Plus, a filter, such as a dichroic filter 240, can be used to filter out any wavelengths of undesired electromagnetic radiation.
When illuminated particles fluoresce, the fluorescence 215 can be reflected by the second reflector back towards a reflective surface, such as dichroic filter 240 which reflects the fluorescence to detector housing 244 to be detected. Because of the ellipsoidal geometry a converging set of beams is created - thus, there is no need for optics to focus the fluorescence on the detector. Figure 8 also shows that a stream of cells can be deflected for sorting or analysation purposes as they fall through an opening in the second reflector 216.
In Figure 8, the first reflector and second reflector are shown having focal lengths of fl and £2 respectively. When these focal lengths are equivalent and the distant focal points are coincident and the central axes are aligned as shown, the real image of the arc lamp will be the same size as the actual arc lamp. However, in some cases it is desirable to shrink the size of the real image of the arc source. This is the case when there is a possibility of two cells being very close to one another in the inspection zone of the stream. In such a case, it can be important to reduce a real image so that incident radiation is incident upon only the cell under analysation and not a second cell nearby. This prevents fluorescence from a second cell which might give an incorrect analysis. There is more likelihood of cells being close by when the throughput of the analyser is increased.
The arrangement of Figure 8 could be used with only the bottom reflector and an alternative light source to illuminate the flow of particles. This might involve a laser directed at the flow of particles or off the reflective surface of the ellipsoidal reflector 216. This is a unique arrangement in flow cytometry, because the flow of particles is aligned coaxially with the central axis of the ellipsoidal reflector 216 to pass through the near focal point of the ellipsoidal reflector 216. After the flow of particles passes through the focal point at which the particles are irradiated with electromagnetic radiation for the purpose of analysation, they can be sorted based upon their identifying characteristics. Electrostatic plates can be provided and disposed below the opening in the ellipsoidal reflective surface to deflect the particles as they pass close to or between the electrostatic plates. This embodiment is particularly unique in jet-in-air types of flow cytometers.
In Figure 9 a similar arrangement to that shown in Figure 8 can be seen, the major difference being that paraboloidal shapes are being used for the reflectors. A first reflector 200 having a partial paraboloidal shape, a focal point (or focus) 302 is disposed to reflect electromagnetic radiation from a source of electromagnetic radiation, such as arc lamp 312. The source of electromagnetic radiation can be positioned at the focus of the paraboloid such that all emissions originating from the focus and incident on the partial paraboloid are reflected in collimated beams 313 toward a second reflector 316. The first reflector 300 and the second reflector 316 each have parabolic axes 308 and 338 respectively. These axes can be aligned such that a real image of the electromagnetic source appears at the focal point (or focus) 320 of the second reflector 316. A flow source 332 can provide a flow of particles 337 that flows through the focal point 320 of the second reflector 316. The portion of the flow of particles that flows through the focal point can be referred to as the inspection zone 336 upon which the electromagnetic radiation is focused so as to analyse a cell falling through the inspection zone.
When the incident electromagnetic radiation is incident upon a cell in the inspection zone, the stained cell can be caused to fluoresce. This fluorescence 315 can then be reflected by the second reflector 316 toward a reflector, such as dichroic mirror 340, which directs the fluorescence toward an optical apparatus 345 that focuses the fluorescence on a detector 344.
Once again, selection of equivalent focal lengths for the first reflector fl and second reflector f2 will provide a real image of the arc lamp of the same size at the focal point of the second reflector. Similarly, choosing a focal length for the second reflector that is smaller than the focal length of the first reflector will result in a smaller image that will help prevent error when large throughput of cells is desired and consequently cells are close together at the inspection zone.
In Figures 8 and 9, one can see that plates can be provided to sort cells as they exit the ellipsoidal or paraboloidal shapes.
In another embodiment of the invention, a nozzle 400 can be provided with a reflector coupled to the nozzle itself. In fact, the reflector can even be integral to the nozzle. This presents a significant advantage to the user of the analysing apparatus as there is no need for alignment of the components since the coupling can accomplish that task. Referring to Figures 10, 11, 12 and 13 one can see how various embodiments of such a nozzle could be implemented. In Figure 10, a paraboloidal nozzle is shown. The nozzle can be manufactured of a material such as glass that permits the transmission of electromagnetic radiation, such as visible light. Incident beams of electromagnetic radiation from a source of electromagnetic radiation, such as a laser source 520 in Figure 11 pass through the nozzle body and are incident on a reflector 402. The reflector 402 is coupled to the nozzle itself rather than existing separate from the nozzle. An opening 404 can be provided in the nozzle to allow a flow of particles 408 to flow through. The reflector 402 can be oriented to reflect the incident electromagnetic radiation at the flow of particles 408.
Two possible shapes which can be used for the reflective surface of the reflector are a paraboloid and an ellipsoid. In Figure 10, a paraboloidal reflective surface 412 is shown while in Figure 11, an ellipsoidal reflective surface 512 is shown. As explained elsewhere, an inspection zone 416 can overlap a focal point(s) of the reflective surface, such as focal point 420 of the paraboloid of Figure 10 to produce the desired reflection patterns.
The nozzle can be used with a source of electromagnetic radiation, such as a laser source 520 as shown in Figure 11. However, it is also envisioned that an arc lamp or other source could be used as well. The source of electromagnetic radiation emits beams 450 which can be directed at the reflective surface. When the electromagnetic radiation is incident upon a cell under analysis, fluorescence is created as shown by beams 451.
To create the reflective surface, a variety of designs are possible. First, the nozzle body could be shaped in a paraboloidal or ellipsoidal shape and then coated with a reflective material 428 applied to the nozzle surface. Additionally, a reflector, such as a metal reflector 424 could be inserted or embedded in the nozzle body as shown in Figure 12. It might even be possible to rely on refractive properties which cause internal reflection or even total internal reflection.
In Figure 13, an embodiment is shown in which the nozzle is shaped such that the focal point 420 of the reflective surface is external to the nozzle. External is intended to means outside of or away from the nozzle border, In such an embodiment, electromagnetic radiation could be directed at the focal point without needing to traverse through the nozzle body.
Alternative embodiments of the invention can be seen in figures 14 and 15. In Figure 14, the radial optics configuration for a flow cytometer 500 can combine 360 degree radial illumination and radially symmetric collection of fluorescence from particles or cells as they pass through the inspection point. A glass cone 516 and a paraboloidal reflector 528 can be used. The optical beam of a laser 508 can be steered onto the point of the glass cone. The beam can then be refracted into a divergent cone of light which is retro-reflected to produce a cylinder of laser light which encircles and is antiparallel to the input beam. This light cylinder can then be reflected by a 45 degree elliptical ring mirror 512 and aligned parallel to the optical axis of the paraboloidal reflector 528. The angle of incidence of the cylindrical beam at the reflector is 45 degrees, causing the beam to form a coplanar convergent disk perpendicular to and focused on the sample stream.
Stained cells can be carried by the sample stream through the radial excitation focus and caused to fluoresce. Much of the fluorescence can be collected by the paraboloidal reflector and projected out in a collimated beam onto an aspheric condensing lens 504. The lens can focus the fluorescent light to a spot which is imaged by a microscope objective 520 into a phomultiplier tube (PMT) 501 and filter housing. Optical alignment of specimens flowing through the focal region of the paraboloidal reflector can be achieved by adjusting the flow cell position to maximise fluorescent signals from calibration microspheres. The paraboloidal reflector can have a hole or opening in the base through which the sample stream can exit and where a jet observation camera and droplet sorting mechanism 532 can be situated.
In Figure 15, a simplified version of the geometry of Figure 14 is shown. The fluorescence collection elements can be retained to provide radially symmetric detection of cells as they pass through the inspection point of the flow cytometer. Excitation of cells can be performed by steering a laser beam 608 onto the paraboloidal reflector 628 at an incidence angle that results in beam delivery from one direction similar to standard flow cytometer illumination. This can be accomplished by reflecting the beam off mirror 612. Detection of cells can be performed by a paraboloidal reflector and aspheric lens combination. A single PMT, for example with a 400LP filter, can be positioned to collect all of the light focused by the aspheric lens. An additional neutral density filter (ND=1.3) can also be used to prevent saturation of the detector even at low PMT amplifier voltages. The embodiment in Figure 15 is particularly useful as it does not require as extensive an alignment of optics as is required in other embodiments. An ellipsoidal collector could also be used to deliver the laser light reflected from an adjusted mirror 612 and to reflect fluoresence to be collected at the PMT. The embodiments in Figures 15 and 16 are particularly advantageous because of the simplistic substantially coaxial alignment of the reflector with the detector.
It should be appreciated that the embodiments described in this description rely on physical arrangements that may not permit total or perfect collection, transmission, symmetry, reflection, alignment, etc. due to physical limitations of mirrors, optics and physical orientation of equipment. In view of these limits, such properties still may be considered at the very least as substantial.
Aspects of the present invention have been described by way of example only and it should be appreciated that modifications and additions may be made thereto without departing from the scope thereof.

Claims

1. An optical apparatus including: a prism having a conical portion with an apex at a forward end of the prism and a central axis extending through the apex of the prism; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct an incident beam of electromagnetic radiation onto the apex of the conical portion in a direction substantially aligned with the central axis of the conical portion; and a reflective surface provided behind the apex of the prism; such that the beam refracted by the prism will be reflected by the reflective surface back through the prism to project from the forward end of the prism as an annular beam of electromagnetic radiation.
2. The optical apparatus as claimed in claim 1 wherein the prism also includes a cylindrical base portion at a rear end thereof which has a cross-section corresponding to the base of the conical portion.
3. An optical apparatus including: a prism having a pyramidal portion with an even number of inclined faces meeting at an apex at a forward end of the prism and a central axis extending through the apex; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct an incident beam of electromagnetic radiation onto the apex of the pyramidal portion in a direction substantially aligned with the central axis of the pyramidal portion; and a reflective surface provided behind the apex of the prism; such that the beam refracted by the prism will be reflected by the reflective surface back through the prism to project from the forward end of the prism as a number of parallel beams.
4. The optical apparatus as claimed in claim 3 wherein the prism further includes a base portion at a rear end thereof which has a cross-section corresponding to the base of the pyramidal portion.
5. The optical apparatus as claimed in any one of the preceding claims wherein the reflective surface is provided at the rear end of the prism.
6. The optical apparatus as claimed in claim 2 or claim 4 wherein the reflective surface is provided at the rear end of the base portion.
7. The optical apparatus as claimed in claim 2 or claim 4 wherein the reflective surface is spaced from the base portion.
8. The optical apparatus as claimed in claim 7 wherein the spacing of the reflective surface from the base portion is adjustable.
9. The optical apparatus as claimed in any one of the preceding claims further including a paraboloidal reflector having an internal paraboloidal-shaped reflective surface, a focus and an optical axis, the reflector being oriented to receive on its reflective surface, the electromagnetic radiation projected from the forward end of the prism.
10. The optical apparatus as claimed in claim 9 further including a second reflector disposed between the prism and the paraboloidal reflector, the second reflector having reflective portions to reflect the incident beam from the source onto the apex of the prism and one or more transmitting portions to transmit the beam(s) projected from the forward end of the prism.
11. An optical apparatus including: an optical configuration adapted to produce an annular beam of electromagnetic radiation having a central axis or plurality of beams of electromagnetic radiation wherein said plurality of beams are evenly spaced from a central axis; and a focussing reflector having an internal reflective surface having an optical axis and one or more foci, the reflector being oriented to substantially receive, onto its reflective surface, the annular beam or the plurality of beams of electromagnetic radiation.
12. The optical apparatus claimed in claim 11 wherein the optical element is a reflective axicon.
13. The optical apparatus as claimed in claim 11 wherein the optical element is a waxicon.
14. The optical apparatus as claimed in any one of claims 11 to 13 wherein the internal reflective surface is paraboloidal in shape.
15. The optical apparatus as claimed in any one of claims 9 to 14 wherein the optical axis of the reflective surface is substantially aligned with the central axis.
16. A flow cytometer including the apparatus as claimed in any one of claims 9 to 15 including a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles substantially through the focus or one of the foci of the reflective surface.
17. The flow cytometer as claimed in claim 16 wherein the flow source is adapted to substantially align the flow with the optical axis of the reflective surface.
18. The flow cytometer as claimed in claim 16 or claim 17 wherein an aperture is provided in the focussing reflector for passage of the flow beyond the paraboloidal reflector.
19. The flow cytometer as claimed in claim 18 wherein the flow source includes a nozzle and wherein electrostatic droplet deflection sorting apparatus is provided below the aperture in the focussing reflector.
20. The flow cytometer as claimed in any one of claims 16 to 19 wherein the source of electromagnetic radiation provides ultra violet light.
21. An optical method including: providing a prism having a conical portion with an apex at the forward end, a central axis extending through the apex and a reflective surface provided behind the apex of the prism; directing an incident beam of electromagnetic radiation onto the apex of the conical portion in a direction substantially aligned with the central axis of the conical portion to produce an annular beam of electromagnetic radiation projecting from the forward end of the prism.
22. An optical method including: providing a prism having a pyramidal portion with an even number of inclined faces meeting at an apex at a forward end of the prism, a central axis extending through the apex and a reflective surface provided behind the apex of the prism; directing an incident beam of electromagnetic radiation onto the apex of the pyramidal portion in a direction substantially aligned with the central axis of the pyramidal portion to produce parallel beams of electromagnetic radiation projecting from the forward end of the prism.
23. The optical method as claimed in claim 21 or 22 further including the use of a paraboloidal reflector having an internal reflective surface of paraboloidal shape, the reflective surface having an optical axis and a focus; orienting the paraboloidal reflector such that it receives, onto its reflective surface, the one or more beams of electromagnetic radiation projected from the forward end of the prism.
24. The method claimed in claim 23 further including: substantially aligning the optical axis of the reflective surface with the central axis of the prism.
25. The method as claimed in claim 23 or 24 further including: directing a flow of particles to be analysed through the focus of the paraboloidal reflector.
26. The method as claimed in claim 25 further including: substantially aligning the flow with the optical axis of the reflective surface.
27. An analysation instrument including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to converge substantially coplanar, substantially radially symmetric electromagnetic radiation towards the inspection zone.
28. The analysation instrument as described in claim 27 and further comprising a central axis along which the flow of particles is directed, wherein an annular disc of electromagnetic radiation is disposed symmetrically relative to the central axis.
29. A method of analysing including: providing a flow of particles to be analysed; directing the flow of particles to be analysed through an inspection zone; converging substantially coplanar, substantially radially symmetric electromagnetic radiation towards the inspection zone.
30. The method of claim 29 and further including disposing an annular disc of electromagnetic radiation symmetrically relative to a central axis along which the flow of particles is directed.
31. An analysation instrument including: a flow source to produce a flow of particles to be analysed; a source of electromagnetic radiation; a reflector adapted to reflect at least a portion of the electromagnetic radiation at the flow of particles to illuminate the flow of particles; an optical configuration including a sensor adapted to sense electromagnetic radiation; wherein the reflector is also adapted to reflect, to the optical configuration, at least a portion of any electromagnetic radiation produced as a result of the illumination of the flow of particles.
32. The analysation instrument of claim 31 wherein the reflector has an internal reflective surface which is paraboloidal in shape.
33. A method of analysing including: providing a flow of particles to be analysed; providing a source of electromagnetic radiation; reflecting with a reflector at least a portion of the electromagnetic radiation to illuminate the flow of particles; reflecting with the reflector at least a portion of any electromagnetic radiation produced from the illumination of the flow of particles; sensing a portion of the electromagnetic radiation produced from the illumination of the flow of particles.
34. The method of claim 33 wherein the reflector has an internal reflective surface which is paraboloidal in shape.
35. A flow cytometer including: a flow source to produce a linear flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement adapted to converge electromagnetic radiation onto the flow at the inspection zone in a substantially radially symmetric manner about the inspection zone; a collector to collect electromagnetic radiation either produced or deflected from the particles in the flow; a processor to derive, from the collected electromagnetic radiation, predetermined information relating to each of at least some of the particles in the flow; and a correlator to correlate the derived information with the associated particle downstream of the inspection zone.
36. The flow cytometer claimed in claim 35 wherein the optical arrangement is adapted to produce a continuous disc of electromagnetic radiation convergent towards the inspection zone.
37. The flow cytometer claimed in claim 35 wherein the optical arrangement is adapted to produce discrete beams of electromagnetic radiation convergent towards the inspection zone.
38. A flow cytometer including: a flow source to produce a linear flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; and an optical arrangement including a focussing reflector having an internal reflective surface with one or more foci, the optical arrangement adapted to converge electromagnetic radiation onto the flow of particles at the inspection zone by reflection from the focussing reflector, the focussing reflector being oriented such that one of the one or more foci is substantially coincident with or located within the inspection zone.
39. The flow cytometer as claimed in claim 38 wherein: the focussing reflector has at least a partial ellipsoidal reflective surface and a first focus and a second focus; an optical axis of the ellipsoidal reflective surface oriented along a line between the first and second focus of the ellipsoidal surface; the flow source being oriented so that the flow of particles passes along the optical axis through the second focus of the ellipsoidal reflective surface.
40. The flow cytometer as claimed in claim 38 wherein the focussing reflector comprises a paraboloidal reflector having an internal reflective surface of paraboloidal shape with an optical axis and one focus.
41. The flow cytometer as claimed in claim 40 wherein the optical axis of the reflective surface is substantially aligned with the flow of particles.
42. The flow cytometer as claimed in any one of claims 38 to 41 wherein an aperture is provided in the focussing reflector for passage of the flow there beyond.
43. The flow cytometer as claimed in any one of claims 38 to 41 further including: a collector to collect electromagnetic radiation either produced or deflected from the particles in the flow; a processor to derive, from the collected electromagnetic radiation, predetermined information relating to each of at least some of the particles in the flow; and a correlator to correlate the derived information with the associated particle downstream of the inspection zone.
44. The flow cytometer as claimed in any one of claims 38 to 43 wherein the flow cytometer is a jet-in-air flow cytometer.
45. A flow cytometer including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles, at the inspection zone; a collector to collect electromagnetic radiation either produced or deflected from the particles, the collector having an internal reflective surface with an optical axis and one or more foci, wherein the collector is oriented such that the flow of particles is substantially aligned with the optical axis.
46. A flow cytometer including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles, at the inspection zone; a collector to collect electromagnetic radiation either produced or deflected from the particles, the collector having an internal reflective surface with an optical axis and one or more foci, wherein the collector is disposed such that one of the one or more foci is substantially coincident or located within the inspection zone; a processor to derive, from the collected electromagnetic radiation, predetermined information relating to each of at least some of the particles in the flow; and a correlator to correlate the derived information with the associated particle downstream of the inspection zone.
47. An analysation instrument including: a first reflector having a partial ellipsoidal shape; a near focal point of the partial ellipsoidal shape of the first reflector; a distant focal point of the partial ellipsoidal shape of the first reflector; a central axis of the partial ellipsoidal shape defined by the near focal point and distant focal point of the partial ellipsoidal shape of the first reflector; a source of electromagnetic radiation disposed at the near focal point of the partial ellipsoidal shape capable of emitting electromagnetic radiation toward the first reflector; a second reflector having a partial ellipsoidal shape oriented relative to the first reflector so as to be capable of receiving electromagnetic radiation reflected by the first reflector; a near focal point of the partial ellipsoidal shape of the second reflector;
a distant focal point of the partial ellipsoidal shape of the second reflector; a central axis of the partial ellipsoidal shape defined by the near focal point and distant focal point of the partial ellipsoidal shape of the second reflector; a flow source to produce a flow of particles to be analysed; and an inspection zone of the flow of particles located at the near focal point of the partial ellipsoidal shape of the second reflector.
48. The analysation instrument as claimed in claim 47 wherein the source of electromagnetic radiation comprises an arc lamp.
49. The analysation instrument as claimed in claim 47 or 48 wherein the distant focal point of the partial ellipsoidal shape of the first reflector and the distant focal point of the partial ellipsoidal shape of the second reflector overlap.
50. The analysation instrument as claimed in claim 47, 48 or 49 and further including: a focal length of the first reflector; a focal length of the second reflector; wherein the focal length of the first reflector and the focal length of the second reflector are equivalent.
51. The analysation instrument as claimed in claim 47, 48 or 49 and further comprising: a focal length of the first reflector; a focal length of the second reflector; wherein the focal length of the first reflector is greater than the focal length of the second reflector.
52. The analysation instrument as claimed in any one of claims 47 to 51 and further including: a filter to filter out a portion of the electromagnetic radiation emitted by the source of electromagnetic radiation.
53. A method of analysing including: utilising a first reflector having a partial ellipsoidal surface with a near focal point and a distant focal point; emitting electromagnetic radiation from a source of electromagnetic radiation positioned at the near focal point of the first reflector; reflecting electromagnetic radiation emitted by the source of electromagnetic radiation from the first reflector; utilising a second reflector having a partial ellipsoidal surface with a near focal point and a distant focal point; providing a flow of particles to be analysed; directing the flow of particles through an inspection zone; positioning the second reflector so that the near focal point of the second reflector overlaps the inspection zone and so that the second reflector is capable of receiving electromagnetic radiation reflected by the first reflector.
54. The method of analysing as claimed in claim 53 and further comprising: utilising an arc lamp as the source of electromagnetic radiation.
55. The method of analysing as claimed in claim 53 or 54 and further comprising overlapping the distant focal point of the first reflector and the distant focal point of the second reflector.
56. The method of analysing as claimed in claim 53, 54 or 55 and further comprising: utilising a first reflector having a focal length equivalent to a focal length of the second reflector.
57. The method of analysing as claimed in claim 53, 54, 55 and further comprising: utilising a first reflector having a focal length longer than a focal length of the second reflector.
58. The method of analysing as claimed in any one of claims 53-57 and further comprising: positioning a filter so as to filter out a portion of the electromagnetic radiation emitted by the source of electromagnetic radiation.
59. An analysation instrument including: a first reflector having a partial paraboloidal shape; a focal point, a parabolic axis and a focal length of the partial paraboloidal shape of the first reflector; a source of electromagnetic radiation disposed at the focal point of the partial paraboloidal shape adapted to emit electromagnetic radiation toward the first reflector; a second reflector having a partial paraboloidal shape oriented relative to the first reflector so as to be capable of receiving electromagnetic radiation reflected by the first reflector; a focal point, a parabolic axis and a focal length of the partial paraboloidal shape of the second reflector; a flow source to produce a flow of particles to be analysed; and an inspection zone of the flow of particles located at the focal point of the partial paraboloidal shape of the second reflector.
60. The analysation instrument as claimed in claim 59 wherein the source of electromagnetic radiation comprises an arc lamp.
61. The analysation instrument as claimed in claim 59 or 60 wherein the parabolic axis of the first paraboloidal shape and the parabolic axis of the second paraboloidal shape are collinear.
62. The analysation instrument as claimed in claim 59, 60 or 61 wherein the focal length of the first reflector and the focal length of the second reflector are equivalent.
63. The analysation instrument as claimed in claim 59, 60 or 61 wherein the focal length of the first reflector is greater than the focal length of the second reflector.
64. The analysation instrument as claimed in anyone of claims 59 to 63 and further including: a filter between the focal point of the first reflector and the focal point of the second reflector.
65. A method of analysing including: utilising a first reflector having a partial paraboloidal surface, a parabolic axis and a focal point; emitting electromagnetic radiation from a source of electromagnetic radiation positioned at the focal point of the first reflector; reflecting electromagnetic radiation emitted by the source of electromagnetic radiation from the first reflector; utilising a second reflector having a partial paraboloidal surface, a parabolic axis and a focal point; providing a flow of particles to be analysed; directing the flow of particles through an inspection zone; positioning the second reflector so that the focal point of the second reflector overlaps the inspection zone and so that the second reflector is capable of receiving electromagnetic radiation reflected by the first reflector.
66. The method of analysing as claimed in claim 65 and further including: utilising an arc lamp as the source of electromagnetic radiation.
67. The method of analysing as claimed in claim 65 or 66 and further including: aligning the parabolic axis of the first reflector with the parabolicaxis of the second reflector such that axes are collinear.
68. The method of analysing as claimed in claim 65, 66 or 67 and wherein the first reflector has a focal length equivalent to a focal length of the second reflector.
70. The method of analysing as claimed in any one of claims 65 to 69 and further including: positioning a filter so as to filter out a portion of the electromagnetic radiation emitted by the source of electromagnetic radiation.
71. A nozzle including: an opening for a flow of particles to flow through; a reflector coupled to the nozzle and oriented to reflect electromagnetic radiation at the flow of particles.
72. The nozzle as claimed in claim 71 wherein the reflector is integral to the nozzle.
73. The nozzle as claimed in claim 71 or 72 wherein the nozzle comprises an ellipsoidal reflective surface.
74. The nozzle as claimed in claim 71 or 72 wherein the nozzle comprises a paraboloidal reflective surface.
75. The nozzle as claimed in claim 71, 72, 73 or 74 and further including: an inspection zone of the flow of particles; and a source of electromagnetic radiation oriented to illuminate the inspection zone of the flow of particles.
76. The nozzle as claimed in claim 75 and further including a focal point of the reflective surface and wherein the focal point of the reflective surface overlaps the inspection zone of the flow of particles.
77. The nozzle as claimed in any one of claims 71 to 76 wherein the reflective surface includes a metal shape embedded in the nozzle.
78. The nozzle as claimed in any one of claims 71 to 76 wherein the reflective surface includes a reflective coating applied to the nozzle.
79. The nozzle as claimed in any one of claims 71 to 78 wherein the nozzle is configured so that a focal point of the reflective surface is located external to the nozzle.
80. The nozzle as claimed in any one of claims 71 to 79 wherein the nozzle is adapted to receive electromagnetic radiation through the nozzle to illuminate the reflector.
81. A method of illuminating a flow of particles, the method including: providing a nozzle having a reflector coupled to the nozzle and oriented to reflect electromagnetic radiation; supplying a flow of particles; directing the flow of particles through the nozzle; reflecting electromagnetic radiation with the reflector toward the flow of particles.
82. The method of illuminating as claimed in claim 81 and further including utilising a nozzle with the reflector integral to the nozzle.
83. The method of illuminating as claimed in claim 81 or 82 and further including utilising a nozzle having an ellipsoidal reflective surface.
84. The method of illuminating as claimed in claim 81 or 82 and further including utilising a nozzle having a paraboloidal reflective surface.
85. The method of illuminating as claimed in claim 81, 82, 83 or 84 and further including: providing a source of electromagnetic radiation oriented to illuminate an inspection zone of the flow of particles.
86. The method of illuminating as claimed in claim 85 and further including orienting the reflector so that a focus of the reflective surface overlaps the inspection zone of the flow of particles.
87. The method of illuminating as claimed in any one of claims 81 to 86 and further including embedding a metal reflective surface in the nozzle.
88. The method of illuminating as claimed in any one of claims 81 to 86 and further including applying a reflective coating to the nozzle.
89. The method of illuminating as claimed in any one of claims 81 to 88 and further including configuring the nozzle so that the focus of the reflective surface is located external to the nozzle.
90. The method of illuminating as claimed in any one of claims 81 to 89 and further including configuring the nozzle so as to receive electromagnetic radiation through the nozzle to illuminate the reflector.
91. A flow cytometer including: a flow source to produce a flow of particles to be analysed, the flow source being adapted to direct the flow of particles through an inspection zone; an optical arrangement including a source of electromagnetic radiation, the optical arrangement adapted to direct electromagnetic radiation onto the flow of particles, at the inspection zone; a partial ellipsoidal collector to collect electromagnetic radiation either produced or deflected from the particles, the collector having an internal reflective surface of partial ellipsoidal shape with two foci and an optical axis oriented along a line between the two foci; the flow source being oriented such that the flow of particles is substantially aligned with the optical axis.
92. The flow cytometer as claimed in claim 91 wherein the flow cytometer is a jet- in-air flow cvtometer.
93. The flow cytometer as claimed in claim 92 including electrostatic plates.
94. A method of flow cytometry including: passing a flow of particles to be analysed through an inspection zone; providing a focussing reflector having one or more foci; converging electromagnetic radiation onto the flow of particles at the inspection zone by reflection from the focussing reflector and aligning the inspection zone with one of the one or more foci.
95. The method as claimed in claim 94 including: providing the focussing reflector with an internal paraboloidal reflective surface.
96. The method of flow cytometry as claimed in claim 94 including: providing the focussing reflector with an ellipsoidal reflective surface, a first focus, a second focus and a central axis oriented along a line between the first focus and the second focus; orienting the reflector so that the flow of particles is substantially aligned with the central axis.
97. The method as claimed in any one of claims 94 to 96 including: providing an opening in the ellipsoidal reflective surface for the flow of particles to flow through.
98. The method as claimed in any one of claims 94 to 97 including: collecting the electromagnetic radiation either produced or deflected from the particles in the flow; processing the collected electromagnetic radiation to derive predetermined information relating to each of at least some of the particles in the flow; and correlating the derived information with the associated particle downstream of the inspection zone.
99. The method of flow cytometry as claimed in any one of claims 94 to 98 and further including utilising the flow cytometer as a jet-in-air flow cytometer.
100. The method as claimed in claim 99 and further including utilising electrostatic plates to sort the flow of particles.
PCT/NZ1998/000009 1997-01-31 1998-02-02 Optical apparatus WO1998034094A1 (en)

Priority Applications (15)

Application Number Priority Date Filing Date Title
AT98901601T ATE298084T1 (en) 1997-01-31 1998-02-02 OPTICAL DEVICE AND METHOD
JP53276398A JP4323571B2 (en) 1997-01-31 1998-02-02 Optical device
EP98901601A EP1017987B1 (en) 1997-01-31 1998-02-02 Optical apparatus and method
NZ337340A NZ337340A (en) 1997-01-31 1998-02-02 Flow cytometer inspection light radially symmetrically converged on linear particle stream
DE69830598T DE69830598T2 (en) 1997-01-31 1998-02-02 OPTICAL DEVICE AND METHOD
US09/355,461 US6819411B1 (en) 1997-01-31 1998-02-02 Optical apparatus
AU57836/98A AU752985B2 (en) 1997-01-31 1998-02-02 Optical apparatus
CA002279574A CA2279574C (en) 1997-01-31 1998-02-02 Optical apparatus
US10/990,648 US7221453B2 (en) 1997-01-31 2004-11-16 Optical apparatus
US11/805,572 US7586604B2 (en) 1997-01-31 2007-05-22 Optical apparatus
US12/555,641 US7929137B2 (en) 1997-01-31 2009-09-08 Optical apparatus
US12/944,308 US8975035B2 (en) 1997-01-31 2010-11-11 Method of analyzing cells
US13/086,223 US8553226B2 (en) 1997-01-31 2011-04-13 Optical apparatus
US13/764,640 US8610892B2 (en) 1997-01-31 2013-02-11 Analyzation instrument
US14/635,910 US20150204772A1 (en) 1997-01-31 2015-03-02 Nozzle and method of illuminating particles

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
NZ314169 1997-01-31
NZ31416997 1997-01-31

Related Child Applications (3)

Application Number Title Priority Date Filing Date
US09355461 A-371-Of-International 1998-02-02
US09/355,461 A-371-Of-International US6819411B1 (en) 1997-01-31 1998-02-02 Optical apparatus
US10/990,648 Continuation US7221453B2 (en) 1997-01-31 2004-11-16 Optical apparatus

Publications (1)

Publication Number Publication Date
WO1998034094A1 true WO1998034094A1 (en) 1998-08-06

Family

ID=19926129

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/NZ1998/000009 WO1998034094A1 (en) 1997-01-31 1998-02-02 Optical apparatus

Country Status (8)

Country Link
US (8) US6819411B1 (en)
EP (4) EP2264427B1 (en)
JP (4) JP4323571B2 (en)
AT (1) ATE298084T1 (en)
AU (1) AU752985B2 (en)
CA (1) CA2279574C (en)
DE (1) DE69830598T2 (en)
WO (1) WO1998034094A1 (en)

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002028311A1 (en) 2000-10-05 2002-04-11 Xy, Inc. System of hysteroscopic insemination of mares
US6372422B1 (en) 1997-12-31 2002-04-16 Colorado State University Through Its Agent Colorado State University Research Foundation And Xy, Inc. Multiple sexed embryo production system for mammals
US6524860B1 (en) 1997-12-31 2003-02-25 Xy, Inc. Methods for improving sheath fluids and collection systems for sex-specific cytometer sorting of sperm
WO2004012837A2 (en) 2002-08-01 2004-02-12 Xy, Inc. Low pressure sperm cell separation system
EP1395374A1 (en) * 2001-05-17 2004-03-10 Cytomation, Inc. Flow cytometer with active automated optical alignment system
WO2004074817A1 (en) * 2003-02-14 2004-09-02 Honeywell International Inc. Particle detection system and method
WO2006073492A3 (en) * 2004-07-30 2007-07-05 Biovigilant Systems Inc Pathogen and particle detector system and method
US7713687B2 (en) 2000-11-29 2010-05-11 Xy, Inc. System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations
US7723116B2 (en) 2003-05-15 2010-05-25 Xy, Inc. Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm
US7758811B2 (en) 2003-03-28 2010-07-20 Inguran, Llc System for analyzing particles using multiple flow cytometry units
US7820425B2 (en) 1999-11-24 2010-10-26 Xy, Llc Method of cryopreserving selected sperm cells
US7833147B2 (en) 2004-07-22 2010-11-16 Inguran, LLC. Process for enriching a population of sperm cells
US7838210B2 (en) 2004-03-29 2010-11-23 Inguran, LLC. Sperm suspensions for sorting into X or Y chromosome-bearing enriched populations
US7929137B2 (en) 1997-01-31 2011-04-19 Xy, Llc Optical apparatus
US8004661B2 (en) 2008-06-30 2011-08-23 Microbix Biosystems Inc. Method and apparatus for sorting cells
JP2012005490A (en) * 2000-05-09 2012-01-12 Xy Llc High purity x-chromosome bearing and y-chromosome bearing populations of spermatozoa
US8486618B2 (en) 2002-08-01 2013-07-16 Xy, Llc Heterogeneous inseminate system
US8634077B2 (en) 2008-10-01 2014-01-21 East Carolina University Methods and systems for optically characterizing a turbid material using a structured incident beam
US8665439B2 (en) 2009-06-30 2014-03-04 Microbix Biosystems, Inc. Method and apparatus for limiting effects of refraction in cytometry
EP2720007A1 (en) * 2012-09-05 2014-04-16 Simmonds Precision Products, Inc. Liquid level gauging system with bubble shedding reflectors
US9013692B2 (en) 2008-06-12 2015-04-21 East Carolina University Flow cytometer apparatus for three dimensional difraction imaging and related methods
US9556416B2 (en) 2011-02-15 2017-01-31 Microbix Biosystems Inc. Methods, systems and apparatus for performing flow cytometry
US9879221B2 (en) 2000-11-29 2018-01-30 Xy, Llc Method of in-vitro fertilization with spermatozoa separated into X-chromosome and Y-chromosome bearing populations
US10101274B2 (en) 2012-12-05 2018-10-16 Genepoc Inc. Optical interrogation device

Families Citing this family (79)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE488247T1 (en) 1998-07-30 2010-12-15 Xy Llc HORSE SYSTEM FOR ARTIFICIAL INSERTION WITHOUT SURGICAL INTERVENTION
DE10202999B4 (en) * 2002-01-26 2004-04-15 Palas Gmbh Partikel- Und Lasermesstechnik Method and device for measuring the size distribution and concentration of particles in a fluid
AU2003250800A1 (en) * 2002-08-02 2004-03-03 Siemens Aktiengesellschaft Evaluation of received useful information by the detection of error concealment
AU2003265471B2 (en) 2002-08-15 2009-08-06 Xy, Llc. High resolution flow cytometer
US7169548B2 (en) 2002-09-13 2007-01-30 Xy, Inc. Sperm cell processing and preservation systems
FR2860869B1 (en) * 2003-10-10 2007-04-20 Optis PORTABLE DEVICE FOR MEASURING THE LIGHT INTENSITY OF AN OBJECT AND USE OF SUCH A DEVICE
US7110192B2 (en) * 2005-01-12 2006-09-19 Dako Denmark A/S System and method for a composite lens for a flow cytometer
DE102005031647A1 (en) * 2005-07-06 2007-01-11 Chromasens Gmbh Illumination device for dark-field illumination for optical test device, has reflector designed with elliptical cross-section
JP5112312B2 (en) 2005-07-15 2013-01-09 バイオヴィジラント システムズ インコーポレイテッド Pathogen and particulate detection system and detection method
WO2007041458A2 (en) * 2005-10-03 2007-04-12 The Salk Institute For Biological Studies Maximal-aperture reflecting objective
DE102005054184B4 (en) * 2005-11-14 2020-10-29 Carl Zeiss Microscopy Gmbh Multispectral lighting device and measuring method
US7671985B1 (en) * 2006-03-13 2010-03-02 Milan Milosevic Device for small spot analysis using fiber optic interfaced spectrometers
WO2007127760A2 (en) * 2006-04-24 2007-11-08 Raintree Scientific Instruments (Shanghai) Corporation Spectroscopic ellipsometers
WO2008019130A2 (en) * 2006-08-04 2008-02-14 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health Wide-area fluorescence detection system for multi-photon microscopy
US20080055755A1 (en) * 2006-08-31 2008-03-06 Searete Llc, A Limited Liability Corporation Of The State Of Delaware Electromagnetic device and method
CN101236150B (en) * 2007-02-02 2012-09-05 深圳迈瑞生物医疗电子股份有限公司 Stream type cell technique instrument opto-electronic sensor and its irradiation unit
US7796253B2 (en) * 2007-04-16 2010-09-14 Kabushiki Kaisha Toshiba Image forming apparatus for forming image on record medium
EP2181317B1 (en) * 2007-08-15 2012-12-05 Malvern Instruments Ltd Broad-range spectrometer
JP5252892B2 (en) * 2007-11-20 2013-07-31 オリンパス株式会社 Optical unit
US8628976B2 (en) 2007-12-03 2014-01-14 Azbil BioVigilant, Inc. Method for the detection of biologic particle contamination
EP2232231A4 (en) 2007-12-04 2015-12-02 Particle Measuring Syst Non-orthogonal particle detection systems and methods
DE202008009859U1 (en) * 2007-12-21 2009-03-05 Berthold Technologies Gmbh & Co. Kg Device for the optional measurement of in particular luminescence and / or fluorescence radiation
JP2009156659A (en) * 2007-12-26 2009-07-16 Olympus Corp Measuring apparatus and method
CN102087197B (en) * 2009-12-08 2014-06-18 龚维燕 Method of coaxial lighting in Coulter micro-hole of full-functional blood analyzer and analyzer therefor
US8289603B2 (en) * 2008-02-05 2012-10-16 Reliant Technologies, Inc. Optical pattern generators using axicon segments
EP2284519B1 (en) * 2008-05-15 2015-12-09 Shimadzu Corporation Biological imaging device
JP5667079B2 (en) * 2008-12-18 2015-02-12 アズビル株式会社 Compact detector for simultaneous detection of particle size and fluorescence
US8251887B2 (en) * 2009-01-24 2012-08-28 Xihe Li Reproductive technology of low dose semen production and in vitro/in vitro fertilization in domestic animals
US8512224B2 (en) * 2009-01-24 2013-08-20 Xy, Llc Method of producing an inseminate
KR101297897B1 (en) * 2009-10-30 2013-08-22 후지츠 프론테크 가부시키가이샤 Illumination optical system of image capturing device
SG10201402041RA (en) * 2010-03-09 2014-07-30 Beckman Coulter Inc Calculate Drop Delay For Flow Cytometry Systems And Methods
US20110223587A1 (en) * 2010-03-11 2011-09-15 Schulman Joseph D Optical particle characterization system
US20110223586A1 (en) * 2010-03-11 2011-09-15 David Karabinus Optical particle characterization system
JP2012026837A (en) * 2010-07-22 2012-02-09 Sony Corp Fine particle measurement instrument
US8729502B1 (en) 2010-10-28 2014-05-20 The Research Foundation For The State University Of New York Simultaneous, single-detector fluorescence detection of multiple analytes with frequency-specific lock-in detection
EP2671065B1 (en) * 2011-02-04 2019-07-10 Cytonome/ST, LLC Particle sorting apparatus and method
US9625810B2 (en) * 2011-03-16 2017-04-18 Kla-Tencor Corporation Source multiplexing illumination for mask inspection
EP2694668A4 (en) * 2011-04-06 2015-04-15 Instant Bioscan Llc Microbial detection apparatus and method
CN102553830B (en) * 2011-12-16 2014-12-03 东南大学 Laser scanning sorting machine
JP2013160672A (en) 2012-02-07 2013-08-19 Sony Corp Optical system for fluorescence detection and fine particle analyzing apparatus
US8879797B2 (en) * 2012-05-25 2014-11-04 Fluid Imaging Technologies, Inc. System and method for total internal reflection enhanced imaging flow cytometry
US9888990B2 (en) 2012-06-06 2018-02-13 Inguran, Llc Methods for use of sex sorted semen to improve genetic management in swine
US9433195B2 (en) 2012-06-06 2016-09-06 Inguran, Llc Methods for increasing genetic progress in a line or breed of swine using sex-selected sperm cells
JP5841498B2 (en) * 2012-06-18 2016-01-13 国立大学法人東京工業大学 Object detection device
CN103575632B (en) 2012-08-09 2015-09-23 深圳开立生物医疗科技股份有限公司 A kind of sheath stream device and blood analyser
DE102012108347A1 (en) * 2012-09-07 2014-03-13 Limo Patentverwaltung Gmbh & Co. Kg Method and device for focusing laser light
BR112015022073B1 (en) 2013-04-12 2020-10-06 Becton, Dickinson And Company AUTOMATED CELL CLASSIFICATION ORGANIZATION
US20160252451A1 (en) * 2013-10-15 2016-09-01 National Institute Of Advanced Industrial Science And Technology Optical measuring device and device having optical system
CN103676436B (en) * 2013-12-20 2016-04-06 海信集团有限公司 A kind of light source shaping methods for projecting and structure
US9551645B2 (en) 2014-07-10 2017-01-24 Kinetic River Corp. Flow cytometry apparatus and methods
KR102241978B1 (en) * 2014-09-11 2021-04-19 삼성전자주식회사 Method of inspecting a surface of an object and optical system for performing the same
DE102015114756B4 (en) * 2014-09-25 2021-07-22 Leica Microsystems Cms Gmbh Mirror device
US10161848B2 (en) 2014-12-12 2018-12-25 Becton, Dickinson And Company Methods for aligning a light source with a flow stream and systems thereof
FR3034578B1 (en) * 2015-03-30 2018-04-27 Horiba Abx Sas METHOD AND DEVICE FOR TRIGGERING IMPULSE LUMINOUS SOURCES
AU2016304897A1 (en) 2015-08-11 2018-01-25 Paul Anthony Iorio Stormwater biofiltration system and method
JP5943366B1 (en) * 2015-08-28 2016-07-05 株式会社サタケ Device with optical unit
WO2017039620A1 (en) 2015-08-31 2017-03-09 Hewlett-Packard Development Company, L.P. Spectral microscope
US9983115B2 (en) 2015-09-21 2018-05-29 Fluid Imaging Technologies, Inc. System and method for monitoring particles in a fluid using ratiometric cytometry
US10502902B2 (en) 2015-09-30 2019-12-10 Sony Corporation Optical communication connector, optical communication cable, and electronic device
WO2017062319A1 (en) 2015-10-05 2017-04-13 Becton, Dickinson And Company Automated drop delay calculation
KR102578808B1 (en) 2016-01-21 2023-09-15 도쿄엘렉트론가부시키가이샤 Foreign matter detection device and foreign matter detection method
US10365198B2 (en) * 2016-04-21 2019-07-30 Malvern Panalytical Limited Particle characterization
DE102016113042A1 (en) * 2016-07-15 2018-01-18 B. Braun Melsungen Ag Flow cell device for measuring fluid parameters
USD868991S1 (en) 2017-03-28 2019-12-03 Becton, Dickinson And Company Register block
USD869676S1 (en) 2017-03-28 2019-12-10 Becton, Dickinson And Company Particle sorting module
DE102017115661A1 (en) * 2017-07-12 2019-01-17 Endress+Hauser Conducta Gmbh+Co. Kg Optical sensor
WO2019064040A1 (en) 2017-09-29 2019-04-04 Nanovare Sas Devices and methods for analysis of male infertility
WO2019138028A1 (en) * 2018-01-11 2019-07-18 Universiteit Hasselt Analysis of data obtained by fluorescence fluctuation microscopy from a plurality of dyes
USD882817S1 (en) 2018-01-30 2020-04-28 Becton, Dickinson And Company Sample container
USD872296S1 (en) 2018-01-30 2020-01-07 Becton, Dickinson And Company Particle sorting module
USD876668S1 (en) 2018-01-30 2020-02-25 Becton, Dickinson And Company Particle sorting module mount
USD864415S1 (en) 2018-01-30 2019-10-22 Becton, Dickinson And Company Particle sorting system
CN112005100A (en) 2018-03-12 2020-11-27 查珀尔希尔北卡罗来纳大学 Optical disc microscopy for fluorescence microscopy
US11262570B2 (en) * 2018-03-12 2022-03-01 The University Of North Carolina At Chapel Hill Mirror image microscopy for increased collection
CN108801883B (en) * 2018-04-19 2021-01-05 深圳市趣方科技有限公司 Micro suspended particle flow optical detection mechanism and detection method
EP3821227B1 (en) 2018-07-10 2024-03-13 Gerrit Jan Van Den Engh System, apparatus and method for off-axis illumination in flow cytometry
WO2022204600A1 (en) 2021-03-26 2022-09-29 Cytonome/St, Llc Systems and methods for particle sorting with automated adjustment of operational parameters
RU2763682C1 (en) * 2021-06-09 2021-12-30 Геннадий Евгеньевич Котковский Optical system for determining aerosol compositions based on luminescent analysis of aerosol particles
WO2023053438A1 (en) 2021-10-01 2023-04-06 株式会社テックジェーピー Thin beam generation device

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4179218A (en) * 1978-05-15 1979-12-18 The Boeing Company Particle size analyzer
US4274740A (en) * 1978-07-21 1981-06-23 Eidenschink Herrn H Optical process and apparatus for determining particle size of colloidal solution
US5132548A (en) * 1990-09-14 1992-07-21 High Yield Technology High sensitivity, large detection area particle sensor for vacuum applications
US5471299A (en) * 1992-02-21 1995-11-28 The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland Apparatus and method for the analysis of particle characteristics using monotonically scattered light

Family Cites Families (667)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US32350A (en) * 1861-05-21 Improvement in machines for turning and spreading hay
US2149735A (en) * 1935-07-08 1939-03-07 Alan W Ross Photosensitive measuring instrument
US3005756A (en) 1958-11-14 1961-10-24 Noland L Van Demark Diluter containing carbon dioxide for preserving semen
US3299354A (en) 1962-07-05 1967-01-17 Coulter Electronics Aperture tube structure for particle study apparatus
US3499435A (en) 1967-06-02 1970-03-10 Paul E Rockwell Esophageal probe for use in monitoring
US3547526A (en) 1967-10-26 1970-12-15 Kollsman Instr Corp Optical beam cross-section converter
US3810010A (en) 1968-11-02 1974-05-07 Telefunken Patent Particle analysis method and apparatus wherein liquid containing particles is sucked into a constricted flow path
US3829216A (en) 1968-11-26 1974-08-13 M Persidsky Optical system and method for counting sperm cells
DE1815352C3 (en) 1968-12-18 1975-03-20 Wolfgang Prof. Dr. Dittrich Automatic measuring and counting device for the particles of a dispersion
US4474875A (en) 1969-04-10 1984-10-02 Wallace Shrimpton Method and means for controlling the sex of mammalian offspring and product therefor
US4327177A (en) 1969-04-10 1982-04-27 Wallace Shrimpton Method and means for controlling the sex of mammalian offspring and product therefor
US3894529A (en) 1969-04-10 1975-07-15 Bio Controls Inc Method and means for controlling the sex of mammalian offspring and product therefor
DE1919628C3 (en) 1969-04-18 1975-04-10 Wolfgang Prof. Dr. Dittrich Arrangement for the automatic counting and / or classification of particles dispersed in a flowable medium
US3687806A (en) 1969-11-04 1972-08-29 Bio Controls Inc Method for controlling sex of mammalian offspring
US3788744A (en) 1970-01-14 1974-01-29 Bio Physics Systems Inc Method and apparatus for photoanalysis
US3661460A (en) 1970-08-28 1972-05-09 Technicon Instr Method and apparatus for optical analysis of the contents of a sheathed stream
US3816249A (en) 1970-11-23 1974-06-11 B Bhattacharya Universal medium and method for extending the useful life of semen in vitro
US3644128A (en) 1970-12-28 1972-02-22 Stuart Lipner Method of preparing comminuted meat products
US3756459A (en) 1971-01-12 1973-09-04 Damon Corp Method and apparatus for metering fluid utilizing pressure differentials
US3916143A (en) 1971-04-22 1975-10-28 Research Corp Branding living animals
US3791384A (en) 1971-07-15 1974-02-12 Schaumann H Artificial insemination of sows
US3833796A (en) 1971-10-13 1974-09-03 Georgia Tech Res Inst Method and apparatus for chromosome digitizing
BE793185A (en) 1971-12-23 1973-04-16 Atomic Energy Commission APPARATUS FOR QUICKLY ANALYZING AND SORTING PARTICLES SUCH AS BIOLOGICAL CELLS
US3826364A (en) 1972-05-22 1974-07-30 Univ Leland Stanford Junior Particle sorting method and apparatus
US3761941A (en) 1972-10-13 1973-09-25 Mead Corp Phase control for a drop generating and charging system
CA1029833A (en) 1973-02-23 1978-04-18 Hildegarde Goehde Apparatus for the automatic counting and measuring of suspended particles
US3791517A (en) 1973-03-05 1974-02-12 Bio Physics Systems Inc Digital fluidic amplifier particle sorter
US4009260A (en) 1973-04-19 1977-02-22 Schering Aktiengesellschaft Fractionation of sperm
USRE29141E (en) 1973-06-14 1977-02-22 Coulter Electronics, Inc. Apparatus for orienting generally flat particles for sensing
US3893766A (en) 1973-06-14 1975-07-08 Coulter Electronics Apparatus for orienting generally flat particles for slit-scan photometry
JPS5023242A (en) * 1973-06-28 1975-03-12
US3947093A (en) * 1973-06-28 1976-03-30 Canon Kabushiki Kaisha Optical device for producing a minute light beam
JPS5613928B2 (en) * 1973-07-13 1981-04-01
US3944917A (en) 1973-08-13 1976-03-16 Coulter Electronics, Inc. Electrical sensing circuitry for particle analyzing device
US3909744A (en) * 1973-09-24 1975-09-30 United Technologies Corp Unstable resonator system producing a high irradiance beam in the far field
US3906929A (en) 1973-11-23 1975-09-23 Lynn Lawrence Augspurger Processes for reproduction of cellular bodies
FR2254639B1 (en) 1973-12-18 1978-06-02 Agronomique Inst Nat Rech
US4070617A (en) 1974-05-08 1978-01-24 Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. Device for controlling the particle flow in an apparatus for measuring the properties of particles suspended in liquid
US3973196A (en) 1974-05-24 1976-08-03 Coulter Electronics, Inc. Method and apparatus for ejecting a metered amount of particulate sample
US3963606A (en) 1974-06-03 1976-06-15 Coulter Electronics, Inc. Semi-automatic adjusting delay for an electronic particle separator
US3877430A (en) 1974-07-17 1975-04-15 Horst K Wieder Artificial insemination apparatus
US4083957A (en) 1974-07-26 1978-04-11 Lang John L Process for the alteration of the sex-ratio of mammals
US4006360A (en) 1974-08-21 1977-02-01 Block Engineering, Inc. Method of discriminating between dyed particles and background fluorescence of the dye
US3920993A (en) * 1974-09-18 1975-11-18 Beckman Instruments Inc Piggyback optical bench
US3999855A (en) * 1974-10-24 1976-12-28 Block Engineering, Inc. Illumination system
USRE32350E (en) 1974-11-22 1987-02-10 Bhairab C. Bhattacharya Thermal convection counter streaming sedimentation and forced convection galvanization method for controlling the sex of mammalian offspring
US4092229A (en) 1975-12-17 1978-05-30 Bhattacharya Bhairab C Thermal convection counter streaming sedimentation and forced convection galvanization method for controlling the sex of mammalian offspring
US3976197A (en) 1974-11-22 1976-08-24 Bhattacharya Bhairab C Thermal convection counter streaming sedimentation method and apparatus for controlling the sex of mammalian offspring
US4014611A (en) 1975-04-30 1977-03-29 Coulter Electronics, Inc. Aperture module for use in particle testing apparatus
US3989381A (en) * 1975-05-05 1976-11-02 Coulter Electronics, Inc. Optical chamber with spherical reflective portion and apparatus employing same
DE2521236C3 (en) 1975-05-10 1978-12-14 Hildegard Dr. 4400 Muenster Goehde Geb. Kuhl Device for counting and measuring particles suspended in a liquid
US3960449A (en) 1975-06-05 1976-06-01 The Board Of Trustees Of Leland Stanford Junior University Measurement of angular dependence of scattered light in a flowing stream
US4058732A (en) 1975-06-30 1977-11-15 Analytical Radiation Corporation Method and apparatus for improved analytical fluorescent spectroscopy
US4021117A (en) * 1975-08-07 1977-05-03 Hildegard Gohde Process for automatic counting and measurement of particles
US4007087A (en) 1975-10-17 1977-02-08 Gametrics Limited Sperm fractionation and storage
AU2154077A (en) 1976-01-27 1978-07-27 Univ Edinburgh Control of sex ratio in mammalian offspring
DE2710030C3 (en) * 1976-03-11 1981-06-19 Novosibirskij Inst Org Chimii Device for photometry of a substance in a cylindrical cuvette
US4162282A (en) 1976-04-22 1979-07-24 Coulter Electronics, Inc. Method for producing uniform particles
US4302166A (en) 1976-04-22 1981-11-24 Coulter Electronics, Inc. Droplet forming apparatus for use in producing uniform particles
JPS5319891A (en) 1976-06-10 1978-02-23 Coulter Electronics Method and apparatus for folling drop formation and separation
GB1583150A (en) 1976-08-02 1981-01-21 Milk Marketing Board Apparatus for collecting eggs
US4110604A (en) 1976-11-04 1978-08-29 Becton, Dickinson And Company Particle density measuring system
DE2709399C3 (en) 1977-03-04 1980-07-24 Goehde, Wolfgang, Dr., 4400 Muenster Device for measuring cell properties
DE2716095A1 (en) 1977-04-12 1978-10-19 Zoeld Tibor Dr Phys GAS CONTROLLED PROCESS FOR SORTING PARTICLES SUSPENDED IN AN ELECTROLYTE AND DEVICE FOR CARRYING OUT THE PROCESS
US4191749A (en) 1977-10-11 1980-03-04 Bryant Bernard J Method and material for increasing the percentage of mammalian offspring of either sex
US4448767A (en) 1977-10-11 1984-05-15 Sumar Corporation Preparation of monospecific male-specific antibody and the use thereof for increasing the percentage of mammalian offspring of either sex
US4189236A (en) 1978-03-20 1980-02-19 Coulter Electronics, Inc. Ellipsoid-conic radiation collector and method
US4188543A (en) * 1978-03-20 1980-02-12 Coulter Electronics, Inc. Ellipsoid radiation collector apparatus and method
US4251733A (en) * 1978-06-29 1981-02-17 Hirleman Jr Edwin D Technique for simultaneous particle size and velocity measurement
US4199686A (en) * 1978-07-31 1980-04-22 Coulter Electronics, Inc. Dark field illuminator and collector apparatus and method
US4276139A (en) 1978-08-16 1981-06-30 Lawson Rommon L Process for magnetic separation and collection of viable female and male spermatozoa
US4225405A (en) 1978-08-16 1980-09-30 Lawson Rommom L Process for separation and collection of viable female and male spermatozoa
US4230558A (en) 1978-10-02 1980-10-28 Coulter Electronics, Inc. Single drop separator
DE3000034A1 (en) * 1979-01-02 1980-07-31 Coulter Electronics METHOD AND DEVICE FOR MEASURING THE DIRECTIONAL DISTRIBUTION PROPERTIES OF THE RADIATION ENERGY REFLECTED BY A PARTICLE
US4341471A (en) 1979-01-02 1982-07-27 Coulter Electronics, Inc. Apparatus and method for measuring the distribution of radiant energy produced in particle investigating systems
SE428972B (en) * 1979-03-07 1983-08-01 Svenska Utvecklings Ab DEVICE FOR DETECTING THE EVENT OF FLUATING, SOLID OR LIQUID PARTICLES IN A GAS
US4267268A (en) 1979-03-12 1981-05-12 Nelson Jr Robert A Spermatozoa extenders
US4274408A (en) 1979-03-26 1981-06-23 Beatrice Nimrod Method for guide-wire placement and novel syringe therefor
US4200802A (en) * 1979-03-28 1980-04-29 The United States Of America As Represented By The United States Department Of Energy Parabolic cell analyzer
NO144002C (en) 1979-04-10 1981-05-27 Norsk Hydro S Inst For Kreftfo DEVICE FOR USE IN LIQUID PHOTOMETRY
US4408877A (en) 1979-04-10 1983-10-11 Ernst Leitz Wetzlar Gmbh Device for hydrodynamic focussing of a particle-suspension in a liquid flow cytophotometer
US4263508A (en) 1979-04-20 1981-04-21 Research Corporation Pulse edge measurement for determining particle dimensional characteristics
US4255021A (en) * 1979-04-20 1981-03-10 The United States Of America As Represented By The United States Department Of Energy Optical device with conical input and output prism faces
JPS5613928A (en) 1979-07-17 1981-02-10 Duskin Franchise Co Method of deodorizing in case of cleaning
US4317520A (en) 1979-08-20 1982-03-02 Ortho Diagnostics, Inc. Servo system to control the spatial position of droplet formation of a fluid jet in a cell sorting apparatus
US4318483A (en) 1979-08-20 1982-03-09 Ortho Diagnostics, Inc. Automatic relative droplet charging time delay system for an electrostatic particle sorting system using a relatively moveable stream surface sensing system
US4318482A (en) 1979-08-20 1982-03-09 Ortho Diagnostics, Inc. Method for measuring the velocity of a perturbed jetting fluid in an electrostatic particle sorting system
US4318480A (en) 1979-08-20 1982-03-09 Ortho Diagnostics, Inc. Method and apparatus for positioning the point of droplet formation in the jetting fluid of an electrostatic sorting device
US4318481A (en) 1979-08-20 1982-03-09 Ortho Diagnostics, Inc. Method for automatically setting the correct phase of the charge pulses in an electrostatic flow sorter
US4325483A (en) 1979-08-20 1982-04-20 Ortho Diagnostics, Inc. Method for detecting and controlling flow rates of the droplet forming stream of an electrostatic particle sorting apparatus
DE2943116C2 (en) 1979-10-25 1986-06-19 Gesellschaft für Strahlen- und Umweltforschung mbH, 8000 München Device for flow cytometric reaction and / or diffusion measurement
US4284355A (en) 1979-10-29 1981-08-18 Ortho Diagnostics, Inc. Automated method for cell volume determination
US4400764A (en) 1980-02-05 1983-08-23 The Boeing Company Low backscatter illumination system
US4367043A (en) 1980-05-05 1983-01-04 Leland Stanford Junior University Method and means for delivering liquid samples to a sample scanning device
US4362246A (en) 1980-07-14 1982-12-07 Adair Edwin Lloyd Method of treating collected mammal semen and separating sperm into X Y components
US4348107A (en) * 1980-07-18 1982-09-07 Coulter Electronics, Inc. Orifice inside optical element
EP0046345A3 (en) 1980-08-15 1982-03-03 Ortho Diagnostic Systems Inc. Controlled hydrodynamic flow in flow cytometry systems
US4350410A (en) 1980-10-08 1982-09-21 The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration Multiprism collimator
US4691829A (en) 1980-11-03 1987-09-08 Coulter Corporation Method of and apparatus for detecting change in the breakoff point in a droplet generation system
US4487320A (en) 1980-11-03 1984-12-11 Coulter Corporation Method of and apparatus for detecting change in the breakoff point in a droplet generation system
US4395676A (en) 1980-11-24 1983-07-26 Coulter Electronics, Inc. Focused aperture module
US4361400A (en) 1980-11-26 1982-11-30 The United States Of America As Represented By The United States Department Of Energy Fluidic assembly for an ultra-high-speed chromosome flow sorter
US4680258A (en) 1981-03-24 1987-07-14 Sloan-Kettering Institute For Cancer Research Process for sex determination in man by use of monoclonal antibodies to the H-Y antigen
US4818103A (en) 1981-05-15 1989-04-04 Ratcom Flow cytometry
US4673288A (en) 1981-05-15 1987-06-16 Ratcom, Inc. Flow cytometry
DE3266669D1 (en) 1981-06-24 1985-11-07 Becton Dickinson Co Analyzer for simultaneously determining volume and light emission characteristics of particles
FR2510393A1 (en) 1981-07-31 1983-02-04 Bertrand Cassou APPARATUS FOR TRANSFERRING ANIMAL REPRODUCTIVE ELEMENTS, SUCH AS EMBRYOS
US4339434A (en) 1981-08-17 1982-07-13 Gametrics Limited Method of increasing the incidence of female offspring
US4395397A (en) 1981-09-17 1983-07-26 Sidney Farber Cancer Institute, Inc. Apparatus and method for killing unwanted cells
US4422761A (en) * 1981-09-28 1983-12-27 Frommer Joseph C Photo-electric particle sensing system
US4515274A (en) 1981-12-02 1985-05-07 Coulter Corporation Particle analyzing and sorting apparatus
SU1056008A1 (en) 1982-01-25 1983-11-23 Предприятие П/Я Р-6681 Flow-type cytofluorimeter
JPS59500340A (en) 1982-03-08 1984-03-01 モトロ−ラ・インコ−ポレ−テツド integrated circuit lead frame
US4511661A (en) 1982-03-19 1985-04-16 University Patents, Inc. ATCC HB8116 And its monoclonal anti-H-Y antibody, Hyclonalan
US4498766A (en) 1982-03-25 1985-02-12 Becton, Dickinson And Company Light beam focal spot elongation in flow cytometry devices
DE3315194A1 (en) 1982-04-29 1983-11-03 International Remote Imaging Systems Inc., 91311 Chatsworth, Calif. METHOD FOR SEPARATING PARTICLES FLOWING IN A FLUID SAMPLE
DE3315195A1 (en) 1982-04-29 1983-11-03 International Remote Imaging Systems Inc., 91311 Chatsworth, Calif. METHOD FOR ALIGNING PARTICLES IN A FLUID SAMPLE
US4629687A (en) 1982-07-29 1986-12-16 Board Of Trustees Of Michigan State University Positive selection sorting of cells
GB2125181B (en) 1982-08-11 1986-01-29 Coulter Electronics Flow cells for particle study
US4559309A (en) 1982-09-01 1985-12-17 Memorial Sloan Kettering Cancer Center Flow cytometry-fluorescence measurements for characterizing sperm
EP0120854A4 (en) 1982-10-05 1985-06-10 Genetic Engineering Inc Method of treating collected mammal semen and separating sperm into x and y components.
FR2535053B1 (en) * 1982-10-22 1985-06-21 Saint Etienne Universite APPARATUS FOR OPTICAL IDENTIFICATION OF INDIVIDUAL MULTIPARAMETRIC PROPERTIES OF PARTICLES OR OBJECTS IN CONTINUOUS FLOW
US4501366A (en) 1982-12-14 1985-02-26 Adolph Coors Company Photomultiplier tube assembly
EP0112188B1 (en) 1982-12-21 1987-06-16 Crosfield Electronics Limited Light beam-splitter
US4492436A (en) 1983-01-03 1985-01-08 At&T Bell Laboratories Polarization independent beam splitter
JPS59143146A (en) 1983-02-07 1984-08-16 Nippon Kogaku Kk <Nikon> Mirror condenser type illuminating optical system
CA1206559A (en) 1983-03-04 1986-06-24 Robert E. Auer Method of and apparatus for detecting change in the breakoff point of a droplet generation system
IT1197570B (en) 1983-02-11 1988-12-06 Serono Ist Farm MIXTURES OF FSH AND LH FROM PORCINE HYPOPHYSIS IN DEFINED RATIO
CH651930A5 (en) 1983-03-24 1985-10-15 Coulter Corp Apparatus for analysis and sorting of particles
JPS59174742A (en) 1983-03-25 1984-10-03 Agency Of Ind Science & Technol Method and apparatus for dividing and sorting fine particle
GB2145112B (en) 1983-04-27 1987-02-18 Milk Marketing Board Sorting living spermatozoa
US4523809A (en) 1983-08-04 1985-06-18 The United States Of America As Represented By The Secretary Of The Air Force Method and apparatus for generating a structured light beam array
NZ207393A (en) 1983-08-05 1987-03-31 Neal Lloyd First Staining dna in living cells
US4538733A (en) 1983-10-14 1985-09-03 Becton, Dickinson And Company Particle sorter with neutralized collection wells and method of using same
US4585736A (en) 1983-10-18 1986-04-29 The United States Of America As Represented By The United States Department Of Energy Flow cytometric measurement of total DNA and incorporated halodeoxyuridine
US4780406A (en) 1983-10-18 1988-10-25 The Regents Of The University Of California Flow cytometric measurement of total DNA and incorporated halodeoxyuridine
US4606636A (en) * 1983-10-25 1986-08-19 Universite De Saint-Etienne Optical apparatus for identifying the individual multiparametric properties of particles or bodies in a continuous flow
JPS6093944A (en) * 1983-10-28 1985-05-25 Satoshi Okuda Light-scattering particle measuring apparatus
US4735504A (en) 1983-10-31 1988-04-05 Technicon Instruments Corporation Method and apparatus for determining the volume & index of refraction of particles
EP0148497B1 (en) 1983-12-24 1990-10-31 Inotech Ag Device for guiding and collecting light in photometry or the like
US4573796A (en) 1984-01-06 1986-03-04 The United States Of America As Represented By The United States Department Of Energy Apparatus for eliminating background interference in fluorescence measurements
US4631483A (en) 1984-02-01 1986-12-23 Coulter Electronics, Inc. Particle analyzing apparatus and method of moving particles in suspension through such apparatus
US4965204A (en) 1984-02-06 1990-10-23 The Johns Hopkins University Human stem cells and monoclonal antibodies
US4714680B1 (en) 1984-02-06 1995-06-27 Univ Johns Hopkins Human stem cells
WO1985004014A1 (en) * 1984-02-29 1985-09-12 Research Corporation Flow cytometers
US4545677A (en) 1984-03-05 1985-10-08 Becton, Dickinson And Company Prismatic beam expander for light beam shaping in a flow cytometry apparatus
US4600302A (en) 1984-03-26 1986-07-15 Becton, Dickinson And Company Flow cytometry apparatus with uniform incoherent light excitation
JPS60205335A (en) * 1984-03-30 1985-10-16 Shimadzu Corp Sheath flow cell device
DE3412620A1 (en) 1984-04-04 1985-10-17 Basf Ag, 6700 Ludwigshafen LASER OPTICAL ARRANGEMENT FOR MEASURING THE DEGREE OF DISPERSION IN FLOWING SYSTEMS
US4609286A (en) 1984-04-16 1986-09-02 Becton, Dickinson And Company Dispersion prism for separation of wavelengths of spectrally rich light in a flow cytometry apparatus
US4605558A (en) 1984-04-20 1986-08-12 Wallace Shrimpton Process for cell separation
US4660971A (en) 1984-05-03 1987-04-28 Becton, Dickinson And Company Optical features of flow cytometry apparatus
FR2563726B1 (en) 1984-05-04 1986-10-10 Robert Cassou ARTIFICIAL INSEMINATION APPARATUS, PARTICULARLY CARNIVORES
FR2566543B1 (en) 1984-06-20 1988-02-26 Commissariat Energie Atomique HIGH COLLECTION AND CYTOFLUORIMETER OPTICAL DEVICE USING THE SAME
EP0171676B1 (en) 1984-07-31 1990-11-07 Hitachi, Ltd. Free-flow electrophoretic separation method and apparatus therefor
US4661913A (en) 1984-09-11 1987-04-28 Becton, Dickinson And Company Apparatus and method for the detection and classification of articles using flow cytometry techniques
EP0177718B1 (en) 1984-09-11 1989-12-06 Partec AG Method and device for sorting microscopic particles
US4598408A (en) 1984-10-22 1986-07-01 Trw Inc. High extraction efficiency cylindrical ring resonator
SU1267231A1 (en) 1984-11-21 1986-10-30 Ленинградский Институт Ядерной Физики Им.Б.П.Константинова Flow-type cytofluorimeter-cell sorter
JPS61139747A (en) 1984-12-12 1986-06-27 Canon Inc Particle analyser
FR2574656B1 (en) 1984-12-13 1988-08-05 Cassou Robert GYNECOLOGICAL PROBE IN PARTICULAR FOR THE INJECTION OF SEMEN OR EMBRYOS INTO THE CAVITY OF ANIMALS, SUCH AS MARES
FR2575063B1 (en) 1984-12-21 1988-07-01 Cassou Robert GYNECOLOGICAL PROBE FOR ARTIFICIAL INSEMINATION, ESPECIALLY FOR SWINE
JPS61159135A (en) 1984-12-31 1986-07-18 Canon Inc Particle analyzing device
SU1260778A1 (en) 1985-01-31 1986-09-30 Центральный научно-исследовательский рентгено-радиологический институт Device for fluorescent analysis of individual microparticles in flow
FR2577317B1 (en) * 1985-02-08 1987-03-06 Inst Rech Hydrologique APPARATUS FOR MEASURING THE TURBIDITY OF A CONTINUOUS FLOW LIQUID.
US4702598A (en) 1985-02-25 1987-10-27 Research Corporation Flow cytometer
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US4683195A (en) 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
CA1250808A (en) 1985-04-29 1989-03-07 David W. Dresser Semen sexing
US4662742A (en) 1985-05-10 1987-05-05 Becton, Dickinson And Company Scatter/fluorescene beam splitter in a flow cytometry apparatus
JPS61294334A (en) 1985-06-21 1986-12-25 Canon Inc Particle analyzer
US4877965A (en) 1985-07-01 1989-10-31 Diatron Corporation Fluorometer
USRE34782E (en) 1985-07-01 1994-11-08 Diatron Corporation Fluorometer
US4744090A (en) 1985-07-08 1988-05-10 Trw Inc. High-extraction efficiency annular resonator
NO156916C (en) 1985-07-10 1987-12-16 Harald B Steen FLOW CLEANING FOR FLUID FLOW PHOTOMETER.
NO156917C (en) 1985-07-16 1987-12-16 Harald B Steen DEVICE FOR MEASUREMENT OF BIOLOGICAL CELLS LIGHT DISTRIBUTION IN LIQUID CURRENT PHOTOMETERS.
US4989977A (en) 1985-07-29 1991-02-05 Becton, Dickinson And Company Flow cytometry apparatus with improved light beam adjustment
US4794086A (en) 1985-11-25 1988-12-27 Liquid Air Corporation Method for measurement of impurities in liquids
US4770992A (en) 1985-11-27 1988-09-13 Den Engh Gerrit J Van Detection of specific DNA sequences by flow cytometry
US4999283A (en) * 1986-01-10 1991-03-12 University Of Kentucky Research Foundation Method for x and y spermatozoa separation
US5756696A (en) 1986-01-16 1998-05-26 Regents Of The University Of California Compositions for chromosome-specific staining
US5447841A (en) 1986-01-16 1995-09-05 The Regents Of The Univ. Of California Methods for chromosome-specific staining
JPS62220833A (en) * 1986-03-22 1987-09-29 Sigma Tec:Kk Light scatter type particulate sensor
US4710635A (en) 1986-04-14 1987-12-01 Becton, Dickinson And Company Dual laser excitation from single laser source
NL8601000A (en) 1986-04-21 1987-11-16 Jan Greve T H Twente Afdeling THE USE OF POLARIZED LIGHT IN FLOW CYTOMETRY.
US5336217A (en) 1986-04-24 1994-08-09 Institut National De La Sante Et De La Recherche Medicale (Insepm) Process for treatment by irradiating an area of a body, and treatment apparatus usable in dermatology for the treatment of cutaneous angio dysplasias
US4790653A (en) 1986-05-22 1988-12-13 Becton Dickinson And Company Housing for a flow cytometry apparatus with particle unclogging feature
JPS62274238A (en) 1986-05-22 1987-11-28 ベクトン・デイツキンソン・アンド・カンパニ− Gel for optical bonding used for flow sight metry device
US4786165A (en) 1986-07-10 1988-11-22 Toa Medical Electronics Co., Ltd. Flow cytometry and apparatus therefor
US4867908A (en) 1986-08-29 1989-09-19 Becton, Dickinson And Company Method and materials for calibrating flow cytometers and other analysis instruments
US4704891A (en) 1986-08-29 1987-11-10 Becton, Dickinson And Company Method and materials for calibrating flow cytometers and other analysis instruments
FR2609885B1 (en) 1987-01-22 1989-04-14 Cassou Robert INSTRUMENT FOR ARTIFICIAL INSEMINATION, TRANSFER OF EMBRYOS OR COLLECTION OF FOLLICULAR LIQUIDS FROM MAMMALS
US4780451B1 (en) 1987-01-23 1995-04-04 Asua International Inc Composition and method for producing superovulation in cattle
US5162306A (en) 1987-01-23 1992-11-10 Donaldson Lloyd E Composition and method for producing superovulation in mammals
EP0279000B1 (en) 1987-02-17 1993-07-21 Ratcom, Inc. Flow cytometry
US4764013A (en) 1987-03-23 1988-08-16 The United States Of America As Represented By The United States Department Of Energy Interferometric apparatus and method for detection and characterization of particles using light scattered therefrom
US4765737A (en) 1987-03-30 1988-08-23 Cornell Research Foundation Cell size measurements using light in flow cytometry and cell sorting
US5021244A (en) 1988-12-06 1991-06-04 Cytogam, Inc. Sex-associated membrane antibodies and their use for increasing the probability that offspring will be of a desired sex
EP0286088B1 (en) 1987-04-08 1994-09-14 Hitachi, Ltd. A sheath flow type flow-cell device
US5346990A (en) 1987-04-08 1994-09-13 Cytogam, Inc. Sex-associated membrane proteins and methods for increasing the probability that offspring will be of a desired sex
JPS63262565A (en) 1987-04-20 1988-10-28 Hitachi Ltd Flow cell
EP0289677A3 (en) 1987-04-27 1989-05-10 Fritz K. Preikschat Apparatus and method for particle analysis
EP0289200B2 (en) 1987-04-27 1998-07-22 Fritz K. Preikschat Apparatus and method for particle analysis
FR2614626B1 (en) * 1987-04-30 1989-07-21 Ranoux Claude CONTAINER FOR FERTILIZATION OF OVOCYTES AND REPLACEMENT OF EMBRYOS IN HUMANS AND ANIMALS
JP2642632B2 (en) 1987-07-03 1997-08-20 株式会社日立製作所 Particle measuring device and particle measuring method
GB8716285D0 (en) 1987-07-10 1987-08-19 Medical Res Council Light collecting device
US4979093A (en) 1987-07-16 1990-12-18 Cavro Scientific Instruments XYZ positioner
US4987539A (en) * 1987-08-05 1991-01-22 Stanford University Apparatus and method for multidimensional characterization of objects in real time
US4796788A (en) 1987-08-26 1989-01-10 Liqui-Box Corporation Bag-in-box packaging and dispensing of substances which will not readily flow by gravity
US4758729A (en) 1987-08-28 1988-07-19 Spectra-Physics, Inc. Apparatus and method for measuring the included angle of a reflective cone
DE3832901A1 (en) 1987-10-02 1989-04-20 Hitachi Ltd PARTICLE MEASURING DEVICE
US4793705A (en) 1987-10-07 1988-12-27 The United States Of America As Represented By The United States Department Of Energy Single molecule tracking
US4831385A (en) 1987-10-14 1989-05-16 Burlington Industries, Inc. Vacuum tray fluid-jet start-up system
US4980277A (en) 1987-10-16 1990-12-25 Cultor Ltd. Cryoprotectant solution and method
US5712807A (en) 1987-10-21 1998-01-27 Bangham; James Andrew Pulse analyzing method and apparatus
US5789155A (en) 1987-10-30 1998-08-04 California Institute Of Technology Process for identifying nucleic acids and triple helices formed thereby
US4845025A (en) 1987-11-10 1989-07-04 Coulter Corporation Biological sample mixing apparatus and method
GB8726304D0 (en) 1987-11-10 1987-12-16 Secr Defence Particle asymmetry analyser
EP0316173A1 (en) 1987-11-10 1989-05-17 The Secretary of State for Defence in Her Britannic Majesty's Government of the United Kingdom of Great Britain and Particle monitoring system
GB8726305D0 (en) 1987-11-10 1987-12-16 Secr Defence Portable particle analysers
US5040890A (en) 1987-11-25 1991-08-20 Becton, Dickinson And Company Sheathed particle flow controlled by differential pressure
US4887721A (en) 1987-11-30 1989-12-19 The United States Of America As Represented By The United States Department Of Energy Laser particle sorter
US4988619A (en) * 1987-11-30 1991-01-29 United States Department Of Energy Flow cytometry apparatus
US4936465A (en) 1987-12-07 1990-06-26 Zoeld Tibor Method and apparatus for fast, reliable, and environmentally safe dispensing of fluids, gases and individual particles of a suspension through pressure control at well defined parts of a closed flow-through system
US5219729A (en) 1988-02-25 1993-06-15 Serono Laboratories, Inc. Fertility assay
US5622820A (en) * 1988-03-10 1997-04-22 City Of Hope Method for amplification and detection of RNA and DNA sequences
US4836038A (en) 1988-03-18 1989-06-06 Aim Instruments Ltd. Automated sampler-injector apparatus and method for sampling a quantity of sample and testing portions of said quantity
US5057413A (en) 1988-06-13 1991-10-15 Becton, Dickinson And Company Method for discriminating between intact and damaged cells in a sample
JPH0213830A (en) 1988-06-30 1990-01-18 Canon Inc Article measuring apparatus
US5070080A (en) 1988-08-10 1991-12-03 Fahim Mostafa S Method of inhibiting generation, maturation, motility and viability of sperm with minerals in bioavailable form
JPH0718785B2 (en) 1988-09-19 1995-03-06 株式会社日立製作所 Flow cell device
JP2635126B2 (en) 1988-09-30 1997-07-30 東亜医用電子株式会社 Particle analysis apparatus and method for determining nuclear leaf index
JP2635125B2 (en) 1988-09-30 1997-07-30 東亜医用電子株式会社 Particle analysis apparatus and method for determining nuclear leaf index
JPH02168160A (en) 1988-12-22 1990-06-28 Omron Tateisi Electron Co Cell selecting apparatus
US5726009A (en) 1989-03-20 1998-03-10 Anticancer, Inc. Native-state method and system for determining viability and proliferative capacity of tissues in vitro
JPH02289808A (en) 1989-04-28 1990-11-29 Olympus Optical Co Ltd Lighting optical system
US4981580A (en) 1989-05-01 1991-01-01 Coulter Corporation Coincidence arbitration in a flow cytomery sorting system
DE69028526T2 (en) 1989-05-10 1997-02-06 Us Agriculture PROCEDURE FOR SELECTING THE GENDER'S GENDER
US4942305A (en) 1989-05-12 1990-07-17 Pacific Scientific Company Integrating sphere aerosol particle detector
DE68924275T2 (en) 1989-05-12 1996-05-02 Cytogam Inc MEMBRANE PROTEINS ASSOCIATED WITH GENDER AND METHODS FOR INCREASING THE LIKELIHOOD THAT THE DEScendants HAVE THE DESIRED GENDER.
FR2647668A1 (en) 1989-06-06 1990-12-07 Medizin Labortechnik Veb K Transfer pipette for embryos
US5055393A (en) 1989-06-13 1991-10-08 Salk Institute Biotechnology/Industrial Associates, Inc. Prenatal sex determination of bovine cells using male-specific oligonucleotides
US4954715A (en) 1989-06-26 1990-09-04 Zoeld Tibor Method and apparatus for an optimized multiparameter flow-through particle and cell analyzer
JPH0353164A (en) 1989-07-20 1991-03-07 Canon Inc Sample supply device and sample measuring instrument using the same
US5098657A (en) 1989-08-07 1992-03-24 Tsi Incorporated Apparatus for measuring impurity concentrations in a liquid
US5030002A (en) 1989-08-11 1991-07-09 Becton, Dickinson And Company Method and apparatus for sorting particles with a moving catcher tube
US5005981A (en) 1989-09-08 1991-04-09 Becton, Dickinson And Company Apparatus for method for causing vortices in a test tube
US5215376A (en) 1989-09-08 1993-06-01 Becton, Dickinson And Company Method for causing vortices in a test tube
EP0418026B1 (en) 1989-09-13 1994-11-30 Kabushiki Kaisha Tiyoda Seisakusho Apparatus for pretreating cells for flow cytometry
JP2808321B2 (en) * 1989-09-19 1998-10-08 東亜医用電子株式会社 Cell analysis method and device
AU6291090A (en) * 1989-09-29 1991-04-28 Atomic Energy Of Canada Limited Infrared-based gas detector
US5072382A (en) 1989-10-02 1991-12-10 Kamentsky Louis A Methods and apparatus for measuring multiple optical properties of biological specimens
US5275787A (en) 1989-10-04 1994-01-04 Canon Kabushiki Kaisha Apparatus for separating or measuring particles to be examined in a sample fluid
EP0422616B1 (en) 1989-10-11 1996-02-07 Canon Kabushiki Kaisha Apparatus for and method of fractionating particle in particle-suspended liquid in conformity with the properties thereof
JPH03140840A (en) 1989-10-26 1991-06-14 Hitachi Ltd Flow cytoanalyser
FR2653885B1 (en) 1989-10-27 1994-01-14 Abx APPARATUS FOR COUNTING AND DETERMINING AT LEAST ONE LEUKOCYTAIC SUB-POPULATION.
US5034613A (en) 1989-11-14 1991-07-23 Cornell Research Foundation, Inc. Two-photon laser microscopy
US5101978A (en) 1989-11-27 1992-04-07 The United States Of America As Represented By The Secretary Of The Army Fluidic sorting device for two or more materials suspended in a fluid
AU647741B2 (en) 1989-12-01 1994-03-31 Regents Of The University Of California, The Methods and compositions for chromosome-specific staining
US5274240A (en) 1990-01-12 1993-12-28 The Regents Of The University Of California Capillary array confocal fluorescence scanner and method
EP0448931B1 (en) 1990-01-26 1996-04-03 Canon Kabushiki Kaisha Method for measuring a specimen by the use of fluorescence light
JP3049254B2 (en) 1990-02-08 2000-06-05 シスメックス株式会社 Optical particle analyzer with two types of light sources
IL93634A0 (en) 1990-03-05 1990-12-23 Galai Lab Ltd Particle size analyzer
EP0446509A1 (en) 1990-03-15 1991-09-18 Panayiotis M. Zavos Semen filter column
US5153117A (en) 1990-03-27 1992-10-06 Genetype A.G. Fetal cell recovery method
US5492534A (en) 1990-04-02 1996-02-20 Pharmetrix Corporation Controlled release portable pump
US5150313A (en) 1990-04-12 1992-09-22 Regents Of The University Of California Parallel pulse processing and data acquisition for high speed, low error flow cytometry
US5076472A (en) 1990-06-13 1991-12-31 Becton, Dickinson And Company Cleaning cycle for flow cytometers
US5087295A (en) 1990-06-13 1992-02-11 Becton Dickinson And Company Cleaning cycle for flow cytometers
JPH0716392B2 (en) 1990-06-20 1995-03-01 株式会社ピーシーシーテクノロジー Cell fusion and fused cell sorting system
JPH0710226B2 (en) 1990-06-20 1995-02-08 株式会社ピーシーシーテクノロジー Protoplast stirring device, cell fusion device and flow cytometer device including the same
US5160974A (en) 1990-06-25 1992-11-03 Flow Science, Inc. Closed sample cell for use in flow cytometry
JPH04126065A (en) 1990-06-25 1992-04-27 Saburo Okonogi Device for molding food raw material into bar-like shape
US5559032A (en) 1990-06-29 1996-09-24 Pomeroy; Patrick C. Method and apparatus for post-transfer assaying of material on solid support
EP0651787A1 (en) 1990-07-09 1995-05-10 Amrad Corporation Limited Enhanced implantation, development and maintenance of embryos using leukaemia inhibitory factor
JP2939647B2 (en) 1990-07-24 1999-08-25 シスメックス株式会社 Automatic focus adjustment method for flow imaging cytometer
IE76732B1 (en) 1990-08-07 1997-11-05 Becton Dickinson Co One step test for absolute counts
US5259593A (en) 1990-08-30 1993-11-09 University Of Southern California Apparatus for droplet stream manufacturing
JPH04115136A (en) 1990-09-05 1992-04-16 Hitachi Ltd Particle measuring apparatus
JPH04126080A (en) 1990-09-14 1992-04-27 Juzo Udaka Production of heat-resistant xylose isomerase
JP2957246B2 (en) 1990-09-14 1999-10-04 大和化成株式会社 Microbial carboxypeptidase B-like enzyme
JPH04126064A (en) 1990-09-14 1992-04-27 Nitto Shokai:Kk Method for preparing doubly covered bean jam food
US5204884A (en) 1991-03-18 1993-04-20 University Of Rochester System for high-speed measurement and sorting of particles
US5273527A (en) 1992-05-12 1993-12-28 Ovamed Corporation Delivery catheter
US5663048A (en) 1990-10-04 1997-09-02 University Of Calgary Y-chromosome specific polynucleotide probes for prenatal sexing
US5840482A (en) * 1990-10-10 1998-11-24 The Regents Of The University Of California Y chromosome specific nucleic acid probe and method for determining the Y chromosome in situ
WO1992008120A1 (en) 1990-10-29 1992-05-14 Macquarie University Pulsed laser flow cytometry
WO1992008122A1 (en) 1990-10-31 1992-05-14 Biophos Medical Ab Fertility analyzer
US5116125A (en) 1990-10-31 1992-05-26 Biophos Medical Ab Fertility analyzer
JP2874746B2 (en) 1990-11-22 1999-03-24 シスメックス株式会社 Flow cell mechanism in flow imaging cytometer
US5991028A (en) 1991-02-22 1999-11-23 Applied Spectral Imaging Ltd. Spectral bio-imaging methods for cell classification
JPH0734012B2 (en) 1991-02-27 1995-04-12 東亜医用電子株式会社 Flow image cytometer
JP3121849B2 (en) 1991-02-27 2001-01-09 シスメックス株式会社 Flow image cytometer
US5144224A (en) 1991-04-01 1992-09-01 Larsen Lawrence E Millimeter wave flow cytometer
US5199576A (en) 1991-04-05 1993-04-06 University Of Rochester System for flexibly sorting particles
EP0515211A3 (en) 1991-05-23 1993-04-07 Becton Dickinson And Company Apparatus and method for phase resolved fluorescence lifetimes of independent and varying amplitude pulses
DE9107792U1 (en) * 1991-06-25 1991-09-12 Labotect-Labor-Technik, Goettingen, Gmbh, 3406 Bovenden, De
FR2678506B1 (en) 1991-07-01 2000-03-10 Claude Ranoux SPONTANEOUS FERTILIZATION PROCESS.
US5548661A (en) 1991-07-12 1996-08-20 Price; Jeffrey H. Operator independent image cytometer
US5412466A (en) 1991-07-26 1995-05-02 Toa Medical Electronics Co., Ltd. Apparatus for forming flattened sample flow for analyzing particles
US5488469A (en) 1991-08-30 1996-01-30 Omron Corporation Cell analyzing apparatus
DE69230902D1 (en) 1991-08-30 2000-05-18 Omron Tateisi Electronics Co Device for cell analysis
US5548395A (en) 1991-09-20 1996-08-20 Toa Medical Electronics Co., Ltd. Particle analyzer
US5578449A (en) 1991-10-03 1996-11-26 Hilding Ohlsson, S.A. Procedure for the sex determination of embryos in mammals especially applied to bovine embryos
IT1253226B (en) 1991-10-24 1995-07-11 Piero Serra CATHETER FOR THE INTRODUCTION OR ASPIRATION OF LIQUIDS OF DIFFERENT NATURES IN ANIMALS PARTICULARLY FOR GYNECOLOGICAL TREATMENTS IN CATTLE, EQUINE AND SIMILAR
JP3212647B2 (en) 1991-10-24 2001-09-25 シスメックス株式会社 Imaging flow cytometer
US5919621A (en) 1991-10-24 1999-07-06 Brown; David B. Methods for diagnosing human male infertility
US5866344A (en) 1991-11-15 1999-02-02 Board Of Regents, The University Of Texas System Antibody selection methods using cell surface expressed libraries
US5370842A (en) 1991-11-29 1994-12-06 Canon Kabushiki Kaisha Sample measuring device and sample measuring system
JP2593021B2 (en) 1991-12-13 1997-03-19 伊藤ハム株式会社 How to identify bovine embryo sex
JP3130628B2 (en) 1992-01-30 2001-01-31 シスメックス株式会社 Particle determination device
US5400179A (en) 1992-02-18 1995-03-21 Asahi Kogaku Kogyo Kabushiki Kaisha Optical multilayer thin film and beam splitter
ATE172791T1 (en) 1992-02-20 1998-11-15 Canon Kk METHOD AND MEASURING APPARATUS FOR HANDLING PARTICLES
US5558998A (en) 1992-02-25 1996-09-24 The Regents Of The Univ. Of California DNA fragment sizing and sorting by laser-induced fluorescence
US6120735A (en) 1992-02-26 2000-09-19 The Ohio States University Fractional cell sorter
JP3203624B2 (en) * 1992-04-30 2001-08-27 株式会社島津製作所 Laser fluorescence detector
US5298967A (en) 1992-06-02 1994-03-29 Pacific Scientific Company Measurement of concentrations of dissolved solvent
JP3145486B2 (en) 1992-06-12 2001-03-12 シスメックス株式会社 Imaging flow cytometer
IT1270457B (en) 1992-07-13 1997-05-05 Pall Corp AUTOMATED SYSTEM AND PROCEDURE FOR THE TREATMENT OF BIOLOGICAL FLUID
JP3215175B2 (en) 1992-08-10 2001-10-02 シスメックス株式会社 Particle analyzer
US5315122A (en) * 1992-08-25 1994-05-24 Becton, Dickinson And Company Apparatus and method for fluorescent lifetime measurement
US5466572A (en) 1992-09-03 1995-11-14 Systemix, Inc. High speed flow cytometric separation of viable cells
ATE219141T1 (en) 1992-09-03 2002-06-15 Systemix Inc THE SEPARATION OF LIVING MAMMAL CELLS BY HIGH-SPEED FLOW CYTOMETRY
US5736410A (en) * 1992-09-14 1998-04-07 Sri International Up-converting reporters for biological and other assays using laser excitation techniques
US5275933A (en) 1992-09-25 1994-01-04 The Board Of Trustees Of The Leland Stanford Junior University Triple gradient process for recovering nucleated fetal cells from maternal blood
US5311290A (en) * 1992-09-30 1994-05-10 Pulp And Paper Research Institute Of Canada Imaging apparatus and method of fiber analysis
US5371585A (en) 1992-11-10 1994-12-06 Pacific Scientific Company Particle detecting instrument with sapphire detecting cell defining a rectangular flow path
US5359907A (en) 1992-11-12 1994-11-01 Horiba Instruments, Inc. Method and apparatus for dry particle analysis
US5395588A (en) 1992-12-14 1995-03-07 Becton Dickinson And Company Control of flow cytometer having vacuum fluidics
FR2699678A1 (en) 1992-12-23 1994-06-24 Unceia Sepn. of mammalian spermatozoa according to sex
EP0679298B1 (en) 1993-01-16 1999-03-31 BANGHAM, James Andrew Signal processing system
JP2525713B2 (en) 1993-01-19 1996-08-21 農林水産省畜産試験場長 Culture and transport method of bovine embryo using low molecular weight thiol compound
US5467189A (en) 1993-01-22 1995-11-14 Venturedyne, Ltd. Improved particle sensor and method for assaying a particle
JP3052665B2 (en) 1993-01-26 2000-06-19 株式会社日立製作所 Flow cell device
CA2113957A1 (en) 1993-01-29 1994-07-30 University Of Guelph Nucleotide sequences for bovine sex determination
EP0682671A4 (en) 1993-02-01 1998-01-14 Seq Ltd Methods and apparatus for dna sequencing.
US5453575A (en) 1993-02-01 1995-09-26 Endosonics Corporation Apparatus and method for detecting blood flow in intravascular ultrasonic imaging
US5367474A (en) 1993-02-08 1994-11-22 Coulter Corporation Flow cytometer
US5547849A (en) 1993-02-17 1996-08-20 Biometric Imaging, Inc. Apparatus and method for volumetric capillary cytometry
US5556764A (en) 1993-02-17 1996-09-17 Biometric Imaging, Inc. Method and apparatus for cell counting and cell classification
US5563059A (en) 1993-02-23 1996-10-08 Genentech, Inc. Use of human inhibin and human activin to increase the number of mature primate oocytes
DE69418131D1 (en) 1993-03-01 1999-06-02 Gen Signal Corp DEVICE FOR GENERATING AN ADJUSTABLE RING-SHAPED LIGHTING FOR A PHOTOLITHOGRAPHIC PROJECTION APPARATUS
NO930980L (en) 1993-03-18 1994-09-19 Flowtech As Optical configuration for fluid flow cytophotometer
US5658751A (en) 1993-04-13 1997-08-19 Molecular Probes, Inc. Substituted unsymmetrical cyanine dyes with selected permeability
US5494795A (en) * 1993-05-05 1996-02-27 The United States Of America As Represented By The Secretary Of The Navy Specific oligonucleotide primers for detection of pathogenic campylobacter bacteria by polymerase chain reaction
US5439578A (en) 1993-06-03 1995-08-08 The Governors Of The University Of Alberta Multiple capillary biochemical analyzer
DE69418248T2 (en) 1993-06-03 1999-10-14 Hamamatsu Photonics Kk Optical laser scanning system with Axikon
CA2140926A1 (en) * 1993-06-04 1994-12-05 Byung H. Chung Artificial insemination and embryo transfer device
NO932088L (en) 1993-06-08 1995-01-05 Oddbjoern Gjelsnes Device for use in liquid flow cytometry
US5483469A (en) 1993-08-02 1996-01-09 The Regents Of The University Of California Multiple sort flow cytometer
US5464581A (en) 1993-08-02 1995-11-07 The Regents Of The University Of California Flow cytometer
US6328071B1 (en) 1993-08-06 2001-12-11 Cary Austin Well pressure tank
US5596401A (en) 1993-09-16 1997-01-21 Toa Medical Electronics Co., Ltd. Particle analyzing apparatus using a coherence lowering device
US5503994A (en) * 1993-10-08 1996-04-02 The Board Of Trustees Of The Leland Stanford Junior University System for sample detection with compensation for difference in sensitivity to detection of components moving at different velocities
US5480774A (en) * 1993-10-14 1996-01-02 A/F Protein, Inc. Determination of genomic sex in salmonids
JP3290786B2 (en) 1993-11-26 2002-06-10 シスメックス株式会社 Particle analyzer
GB9324938D0 (en) 1993-12-04 1994-01-26 Atomic Energy Authority Uk Aerosol generator
FI96452C (en) 1994-01-26 1996-06-25 Pekka Haenninen Method for excitation of dyes
BR9408370A (en) 1994-02-28 1997-08-26 Sunkyong Ind Ltd Pyrimidine acyclonucleoside derivatives
US5475487A (en) 1994-04-20 1995-12-12 The Regents Of The University Of California Aqueous carrier waveguide in a flow cytometer
DE4414940C2 (en) 1994-04-28 1998-07-02 Pekka Haenninen Luminescence scanning microscope with two photons excitation
US5595866A (en) 1994-05-27 1997-01-21 Methodist Hospital Of Indiana, Inc. Step-wise method to remove cryoprotectant from sperm
DE4419894A1 (en) * 1994-06-07 1995-12-14 Gip Medizin Technik Gmbh Endoscopic puncture needle with elastic catheter
EP0687956B2 (en) 1994-06-17 2005-11-23 Carl Zeiss SMT AG Illumination device
US5601234A (en) 1994-08-01 1997-02-11 Abbott Laboratories Fluid nozzle and method of introducing a fluid
US5891734A (en) * 1994-08-01 1999-04-06 Abbott Laboratories Method for performing automated analysis
JP3375203B2 (en) 1994-08-08 2003-02-10 シスメックス株式会社 Cell analyzer
FR2723735B1 (en) 1994-08-18 1996-10-31 Abx Sa AUTOMATIC CONNECTION BOX FOR DISPENSING REAGENTS IN AN APPARATUS IN PARTICULAR A HEMATOLOGICAL ANALYZER.
US20020186874A1 (en) 1994-09-07 2002-12-12 Jeffrey H. Price Method and means for image segmentation in fluorescence scanning cytometry
US5790692A (en) 1994-09-07 1998-08-04 Jeffrey H. Price Method and means of least squares designed filters for image segmentation in scanning cytometry
US5540020A (en) 1994-09-26 1996-07-30 Santini; Daniel E. Building panel
US5700692A (en) 1994-09-27 1997-12-23 Becton Dickinson And Company Flow sorter with video-regulated droplet spacing
US5934885A (en) 1994-10-07 1999-08-10 Bayer Corporation Reagent pump assembly
IL115327A (en) 1994-10-07 2000-08-13 Bayer Ag Diaphragm pump
US5643796A (en) 1994-10-14 1997-07-01 University Of Washington System for sensing droplet formation time delay in a flow cytometer
US5602039A (en) * 1994-10-14 1997-02-11 The University Of Washington Flow cytometer jet monitor system
WO1996012171A2 (en) 1994-10-14 1996-04-25 University Of Washington High speed flow cytometer droplet formation system
US6861265B1 (en) 1994-10-14 2005-03-01 University Of Washington Flow cytometer droplet formation system
US5602349A (en) 1994-10-14 1997-02-11 The University Of Washington Sample introduction system for a flow cytometer
JP3347495B2 (en) 1994-11-14 2002-11-20 シスメックス株式会社 Particle analyzer
US5495719A (en) 1994-11-14 1996-03-05 Gray, Jr.; Carl O. Method of preserving spermatozoa
US5514537A (en) * 1994-11-28 1996-05-07 Board Of Supervisors Of Louisiana State University And Agricultural And Mechanical College Process and apparatus for sorting spermatozoa
WO1996018205A1 (en) 1994-12-08 1996-06-13 Molecular Dynamics, Inc. Fluorescence imaging system employing a macro scanning objective
US5632754A (en) * 1994-12-23 1997-05-27 Devices For Vascular Intervention Universal catheter with interchangeable work element
EP0720012B1 (en) 1994-12-26 2004-09-08 Sysmex Corporation Flow cytometer
US5835262A (en) 1994-12-28 1998-11-10 Research Development Corporation Of Japan Multi-wavelength optical microscope
FI98765C (en) * 1995-01-16 1997-08-11 Erkki Soini Flow cytometric method and apparatus
US5608519A (en) 1995-03-20 1997-03-04 Gourley; Paul L. Laser apparatus and method for microscopic and spectroscopic analysis and processing of biological cells
US5793485A (en) 1995-03-20 1998-08-11 Sandia Corporation Resonant-cavity apparatus for cytometry or particle analysis
US5620842A (en) 1995-03-29 1997-04-15 Becton Dickinson And Company Determination of the number of fluorescent molecules on calibration beads for flow cytometry
US5786560A (en) 1995-03-31 1998-07-28 Panasonic Technologies, Inc. 3-dimensional micromachining with femtosecond laser pulses
US5699152A (en) * 1995-04-03 1997-12-16 Alltrista Corporation Electro-optical inspection system and method
US5528045A (en) 1995-04-06 1996-06-18 Becton Dickinson And Company Particle analyzer with spatially split wavelength filter
US5682038A (en) 1995-04-06 1997-10-28 Becton Dickinson And Company Fluorescent-particle analyzer with timing alignment for analog pulse subtraction of fluorescent pulses arising from different excitation locations
US5641457A (en) 1995-04-25 1997-06-24 Systemix Sterile flow cytometer and sorter with mechanical isolation between flow chamber and sterile enclosure
US5687727A (en) 1995-05-01 1997-11-18 Danforth Biomedical Incorporated Catheter adaptor with slitting blade and improved manual control and method of use
EP0957797A4 (en) 1995-05-09 2001-05-16 Univ Missouri A system for introducing a fluid into the uterus of an animal
FR2734637B1 (en) 1995-05-24 1997-08-14 Abx Sa DEVICE FOR OPTICAL INSPECTION OF A FLUID, PARTICULARLY FOR HEMATOLOGICAL ANALYSIS
US5798276A (en) 1995-06-07 1998-08-25 Molecular Probes, Inc. Reactive derivatives of sulforhodamine 101 with enhanced hydrolytic stability
US5650847A (en) 1995-06-14 1997-07-22 Erkki Soini Method and device for determination of parameters of individual microparticles
US6454945B1 (en) 1995-06-16 2002-09-24 University Of Washington Microfabricated devices and methods
US5716852A (en) 1996-03-29 1998-02-10 University Of Washington Microfabricated diffusion-based chemical sensor
US5589457A (en) 1995-07-03 1996-12-31 Ausa International, Inc. Process for the synchronization of ovulation
US6411835B1 (en) 1997-01-13 2002-06-25 Medispectra, Inc. Spectral volume microprobe arrays
AU722913B2 (en) 1995-08-11 2000-08-17 University Of Guelph Method for identifying sex specific and species specific molecules, molecules identified using the method, and uses of the molecules
AU7237896A (en) 1995-09-06 1997-03-27 Research Foundation Of The State University Of New York, The Two-photon upconverting dyes and applications
DE19533092A1 (en) 1995-09-07 1997-03-13 Basf Ag Device for parallelized two-photon fluorescence correlation spectroscopy (TPA-FCS) and its use for drug screening
US6329158B1 (en) 1995-09-15 2001-12-11 Becton Dickinson And Company Use of dimly fluorescing nucleic acid dyes in the identification of nucleated cells
US6040139A (en) 1995-09-19 2000-03-21 Bova; G. Steven Laser cell purification system
US5726751A (en) 1995-09-27 1998-03-10 University Of Washington Silicon microchannel optical flow cytometer
US5780230A (en) 1995-10-06 1998-07-14 Coriell Institute For Medical Research Compositions and methods for human sperm activation and quantitative assessment of human sperm genome replication
US5736330A (en) 1995-10-11 1998-04-07 Luminex Corporation Method and compositions for flow cytometric determination of DNA sequences
US6117068A (en) 1995-10-19 2000-09-12 Elite Genetics, Inc Artificial insemination system
CA2235069C (en) 1995-10-19 2010-12-14 Advanced Reproduction Technologies, Inc. Methods and compositions to improve germ cell and embryo survival and function
DE69529538T2 (en) 1995-11-09 2003-11-06 Medical Res Council London USE OF CYB MEDIUM FOR TRANSPORTING AND STORING SPERM
DE19549015C1 (en) 1995-12-28 1997-04-03 Siemens Ag Method of monitoring precise location at which fluid stream breaks up into droplets
JP3584108B2 (en) 1996-01-08 2004-11-04 キヤノン株式会社 Lens barrel
US6641708B1 (en) 1996-01-31 2003-11-04 Board Of Regents, The University Of Texas System Method and apparatus for fractionation using conventional dielectrophoresis and field flow fractionation
WO1997029354A1 (en) 1996-02-05 1997-08-14 Bayer Aktiengesellschaft Process and device for sorting and for extraction of biological objects arranged on planar means, such as biological cells or cell organelles, histological sections, chromosome particles etc. using laser beams
BR9600722A (en) 1996-02-14 1997-12-30 Jorge Antonio Rodrigues Claro Infusion pump for contained in flexible plastic bags
WO1997030338A1 (en) 1996-02-16 1997-08-21 Inphocyte, Inc. System and method for rapid analysis of cells using spectral cytometry
JP3127111B2 (en) 1996-02-22 2001-01-22 株式会社日立製作所 Flow type particle image analysis method and apparatus
US6143901A (en) 1996-07-31 2000-11-07 Genesoft, Inc. Complex formation between dsDNA and pyrrole imidazole polyamides
US6090947A (en) 1996-02-26 2000-07-18 California Institute Of Technology Method for the synthesis of pyrrole and imidazole carboxamides on a solid support
US5895922A (en) * 1996-03-19 1999-04-20 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of National Defence Fluorescent biological particle detection system
US5701012A (en) 1996-03-19 1997-12-23 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of National Defence Fluorescent biological particle detection system
US5747349A (en) 1996-03-20 1998-05-05 University Of Washington Fluorescent reporter beads for fluid analysis
JP3640461B2 (en) 1996-04-03 2005-04-20 シスメックス株式会社 Particle analyzer
US5707808A (en) 1996-04-15 1998-01-13 The Regents Of The University Of California Optical selection and collection of DNA fragments
EP1021554B1 (en) 1996-04-25 2007-03-21 Genicon Sciences Corporation Analyte assay using particulate labels
WO1997043620A1 (en) 1996-05-15 1997-11-20 International Remote Imaging Systems, Inc. Selectively emphasizing particles of interest from a fluid sample for analysis
JP2963393B2 (en) * 1996-06-14 1999-10-18 浜松ホトニクス株式会社 Voltage divider circuit for photomultiplier tube
US5846737A (en) 1996-07-26 1998-12-08 Molecular Probes, Inc. Conjugates of sulforhodamine fluorophores with enhanced fluorescence
US5909278A (en) 1996-07-29 1999-06-01 The Regents Of The University Of California Time-resolved fluorescence decay measurements for flowing particles
US5719667A (en) 1996-07-30 1998-02-17 Bayer Corporation Apparatus for filtering a laser beam in an analytical instrument
US5745308A (en) 1996-07-30 1998-04-28 Bayer Corporation Methods and apparatus for an optical illuminator assembly and its alignment
US6074827A (en) 1996-07-30 2000-06-13 Aclara Biosciences, Inc. Microfluidic method for nucleic acid purification and processing
DE69739456D1 (en) 1996-07-30 2009-07-30 Siemens Healthcare Diagnostics Optical system for blood analyzer
US5844685A (en) 1996-07-30 1998-12-01 Bayer Corporation Reference laser beam sampling apparatus
US5872627A (en) 1996-07-30 1999-02-16 Bayer Corporation Method and apparatus for detecting scattered light in an analytical instrument
US6042249A (en) 1996-07-30 2000-03-28 Bayer Corporation Illuminator optical assembly for an analytical instrument and methods of alignment and manufacture
US5883378A (en) 1996-07-30 1999-03-16 Bayer Corporation Apparatus and methods for transmitting electrical signals indicative of optical interactions between a light beam and a flowing suspension of particles
EP0822401A3 (en) 1996-07-30 1999-05-06 Bayer Corporation Hydraulic system for a hematology analytical instrument
US5998140A (en) 1996-07-31 1999-12-07 The Scripps Research Institute Complex formation between dsDNA and oligomer of cyclic heterocycles
US5804436A (en) 1996-08-02 1998-09-08 Axiom Biotechnologies, Inc. Apparatus and method for real-time measurement of cellular response
CA2261148A1 (en) 1996-08-07 1998-02-12 Robert Slovak Additive dispensing apparatus
EP0925494B1 (en) 1996-09-04 2001-12-19 Scandinavian Micro Biodevices A/S A micro flow system for particle separation and analysis
US5873254A (en) * 1996-09-06 1999-02-23 Interface Multigrad Technology Device and methods for multigradient directional cooling and warming of biological samples
AU721792B2 (en) 1996-09-06 2000-07-13 Medarex, Inc. Cyanidin compositions and therapeutic and diagnostic uses therefor
US6221654B1 (en) 1996-09-25 2001-04-24 California Institute Of Technology Method and apparatus for analysis and sorting of polynucleotides based on size
US5759767A (en) 1996-10-11 1998-06-02 Joseph R. Lakowicz Two-photon and multi-photon measurement of analytes in animal and human tissues and fluids
US6002471A (en) 1996-11-04 1999-12-14 California Institute Of Technology High resolution scanning raman microscope
US5799830A (en) 1996-11-08 1998-09-01 Carroll; David C. Pressure vessel access port
US5696157A (en) 1996-11-15 1997-12-09 Molecular Probes, Inc. Sulfonated derivatives of 7-aminocoumarin
AU752985B2 (en) 1997-01-31 2002-10-03 Xy, Llc. Optical apparatus
US6753161B2 (en) 1997-03-27 2004-06-22 Oncosis Llc Optoinjection methods
US5874266A (en) 1997-03-27 1999-02-23 Palsson; Bernhard O. Targeted system for removing tumor cells from cell populations
US6534308B1 (en) * 1997-03-27 2003-03-18 Oncosis, Llc Method and apparatus for selectively targeting specific cells within a mixed cell population
GB9707096D0 (en) 1997-04-08 1997-05-28 Smithkline Beecham Plc Novel device
JP2968231B2 (en) 1997-04-11 1999-10-25 株式会社荏原製作所 Air conditioning system
US6133995A (en) 1997-05-09 2000-10-17 Sysmex Corporation Particle measuring apparatus
US6050935A (en) * 1997-05-09 2000-04-18 Biofertec Container assembly for intravaginal fertilization and culture and embryo transfer and method of intravaginal fertilization and culture employing such a container
CA2293426C (en) 1997-06-09 2007-05-15 Guava Technologies, Inc. Method and apparatus for detecting microparticles in fluid samples
AU8148898A (en) 1997-06-23 1999-01-04 Luminex Corporation Interlaced lasers for multiple fluorescence measurement
ATE326228T1 (en) 1997-07-01 2006-06-15 Vicam Lp METHOD FOR SEX DETERMINATION OF MAMMAL OFFSPRING
US6111398A (en) 1997-07-03 2000-08-29 Coulter International Corp. Method and apparatus for sensing and characterizing particles
BR9704313A (en) 1997-07-08 1999-04-06 Alves Elias Walter Use of chicken egg ig-y associated with monoclonal antibodies against sex specific antigens in bovine sperm immunosexation
US5899848A (en) 1997-07-14 1999-05-04 Haubrich; Mark A. Device and process for artificial insemination of animals
US5985216A (en) 1997-07-24 1999-11-16 The United States Of America, As Represented By The Secretary Of Agriculture Flow cytometry nozzle for high efficiency cell sorting
US5876942A (en) * 1997-07-24 1999-03-02 National Science Council Of Republic Of China Process for sexing cow embryos
US5985538A (en) 1997-08-01 1999-11-16 Saint Barnabas Medical Center Cryopreservation and cell culture medium comprising less than 50 mM sodium ions and greater than 100 mM choline salt
US6003678A (en) 1997-08-21 1999-12-21 University Of Washington Particle separating apparatus and method
US5819948A (en) 1997-08-21 1998-10-13 Van Den Engh; Gerrit J. Particle separating apparatus and method
US5880474A (en) 1997-08-29 1999-03-09 Becton Dickinson And Company Multi-illumination-source flow particle analyzer with inter-location emissions crosstalk cancelation
CA2304197C (en) 1997-09-22 2009-02-03 University Of Guelph Reduction of sperm sensitivity to chilling
US6540895B1 (en) 1997-09-23 2003-04-01 California Institute Of Technology Microfabricated cell sorter for chemical and biological materials
JP3735190B2 (en) * 1997-10-28 2006-01-18 オリンパス株式会社 Scanning cytometer
US6322901B1 (en) 1997-11-13 2001-11-27 Massachusetts Institute Of Technology Highly luminescent color-selective nano-crystalline materials
AU754775B2 (en) 1997-11-19 2002-11-21 University Of Washington High throughput optical scanner
US6086574A (en) 1997-11-21 2000-07-11 Hyclone Laboratories, Inc. Fluid delivery systems with diptube connector
US5895764A (en) * 1997-11-24 1999-04-20 University Of New Mexico Controlled sheath flow injection cytometry
US6207392B1 (en) 1997-11-25 2001-03-27 The Regents Of The University Of California Semiconductor nanocrystal probes for biological applications and process for making and using such probes
US5990479A (en) 1997-11-25 1999-11-23 Regents Of The University Of California Organo Luminescent semiconductor nanocrystal probes for biological applications and process for making and using such probes
EP1040340A4 (en) 1997-12-12 2008-01-09 Chemunex S A Digital flow cytometer
US6149867A (en) * 1997-12-31 2000-11-21 Xy, Inc. Sheath fluids and collection systems for sex-specific cytometer sorting of sperm
US6071689A (en) 1997-12-31 2000-06-06 Xy, Inc. System for improving yield of sexed embryos in mammals
EP1058688A4 (en) 1998-02-03 2004-10-06 Xy Inc Specific oligonucleotide primers for detection of bovine male chromosome presence by polymerase chain reaction and method
CA2321075C (en) 1998-02-20 2009-10-06 Cytomation, Inc. A vibratory system for a sorting flow cytometer
US6746873B1 (en) 1998-02-20 2004-06-08 Xy, Inc. Vibratory system for a sorting flow cytometer
US6154276A (en) 1998-02-23 2000-11-28 The Regents Of The University Of California Waveguide detection of right-angle-scattered light in flow cytometry
US6211477B1 (en) 1998-02-26 2001-04-03 Becton Dickinson And Company Electrostatic deceleration system for flow cytometer
US6248590B1 (en) 1998-02-27 2001-06-19 Cytomation, Inc. Method and apparatus for flow cytometry
US6042025A (en) * 1998-03-13 2000-03-28 Smith Et Al. Two hole dispenser with baffles
EP1064531B1 (en) 1998-03-16 2003-11-19 Partec Partikelzählgeräte GmbH Electronic apparatus for dispensing precise small quantities of fluid
JP3594794B2 (en) 1998-03-24 2004-12-02 独立行政法人 科学技術振興機構 Nanosecond time-gated spectroscopic diagnostic equipment
US6642018B1 (en) 1998-03-27 2003-11-04 Oncosis Llc Method for inducing a response in one or more targeted cells
JP2002510045A (en) 1998-03-30 2002-04-02 バイオシャフ リミテッド Flow-based cell count calculator for analyzing common diagnostic factors in cells and body fluids
US6175409B1 (en) * 1999-04-02 2001-01-16 Symyx Technologies, Inc. Flow-injection analysis and variable-flow light-scattering methods and apparatus for characterizing polymers
JP3350442B2 (en) 1998-04-09 2002-11-25 科学技術振興事業団 Microscope system
JP4215397B2 (en) 1998-05-14 2009-01-28 ルミネックス コーポレイション Multiple analyte diagnostic system
WO1999057955A1 (en) 1998-05-14 1999-11-18 Luminex Corporation Zero dead time architecture and method for flow cytometer
WO1999060397A1 (en) 1998-05-18 1999-11-25 University Of Washington Liquid analysis cartridge
JP2002528699A (en) 1998-05-22 2002-09-03 カリフォルニア インスティチュート オブ テクノロジー Microfabricated cell sorter
US6079836A (en) 1998-07-20 2000-06-27 Coulter International Corp. Flow cytometer droplet break-off location adjustment mechanism
ATE488247T1 (en) 1998-07-30 2010-12-15 Xy Llc HORSE SYSTEM FOR ARTIFICIAL INSERTION WITHOUT SURGICAL INTERVENTION
US6729369B2 (en) 1998-07-31 2004-05-04 Chata Biosystems, Inc. Vessel for containing/transporting a fluent substance
US20040107150A1 (en) 1998-07-31 2004-06-03 Chata Biosystems, Inc. Of Colorado Apparatus and method with vessel for containing/transporting a fluent substance
DE29813921U1 (en) 1998-08-04 1998-10-08 Kisfeld Alfons Device for inseminating animals, especially sows
US6309815B1 (en) 1998-08-07 2001-10-30 University Of Kansas Medical Center Composition and method for preparation, storage and activation of large populations of immotile sperm
AU5675099A (en) 1998-08-17 2000-03-06 William E. Trout Use of zeranol to modulate reproductive cycles
JP2002523738A (en) * 1998-08-21 2002-07-30 ユニオン バイオメトリカ インコーポレイテッド Multicellular organisms and other large object selection and storage devices
US6326144B1 (en) 1998-09-18 2001-12-04 Massachusetts Institute Of Technology Biological applications of quantum dots
US6495333B1 (en) 1998-09-22 2002-12-17 Becton Dickinson And Company Flow cytometric, whole blood dendritic cell immune function assay
US6208411B1 (en) 1998-09-28 2001-03-27 Kla-Tencor Corporation Massively parallel inspection and imaging system
US6707555B1 (en) 1998-10-15 2004-03-16 Sysmex Corporation Optical information measuring apparatus
US6423505B1 (en) 1998-12-03 2002-07-23 Becton Dickinson And Company Methods and reagents for quantitation of HLA-DR and CD11b expression on peripheral blood cells
US7649153B2 (en) * 1998-12-11 2010-01-19 International Business Machines Corporation Method for minimizing sample damage during the ablation of material using a focused ultrashort pulsed laser beam
US7116407B2 (en) 1998-12-15 2006-10-03 Union Biometrica, Inc. System for axial pattern analysis of multicellular organisms
US6128133A (en) 1998-12-22 2000-10-03 Lucent Technologies Inc. Optical beamsplitter
JP2000206032A (en) 1999-01-06 2000-07-28 Bayer Corp Counting method for variable-ratio-volume particles and method for decision of characteristic of sample
US6256096B1 (en) 1999-01-11 2001-07-03 Softray Flow cytometry apparatus and method
US20010006416A1 (en) 1999-01-11 2001-07-05 Johnson Paul E. Ribbon flow cytometry apparatus and methods
US6150119A (en) 1999-01-19 2000-11-21 Caliper Technologies Corp. Optimized high-throughput analytical system
US6580504B1 (en) 1999-01-25 2003-06-17 Amnis Corporation Multipass cavity for illumination and excitation of moving objects
US6473176B2 (en) 1999-01-25 2002-10-29 Amnis Corporation Imaging and analyzing parameters of small moving objects such as cells
US6707551B2 (en) * 2000-01-24 2004-03-16 Amnis Corporation Multipass cavity for illumination and excitation of moving objects
US6671044B2 (en) 1999-01-25 2003-12-30 Amnis Corporation Imaging and analyzing parameters of small moving objects such as cells in broad flat flow
US6119465A (en) 1999-02-10 2000-09-19 Mullens; Patrick L. Shipping container for storing materials at cryogenic temperatures
FR2789778B1 (en) * 1999-02-12 2001-09-14 France Telecom METHOD FOR ASSOCIATING ROUTING REFERENCES WITH DATA PACKETS BY MEANS OF A SORTED MEMORY, AND PACKET ROUTER APPLYING THIS METHOD
US6323632B1 (en) 1999-08-13 2001-11-27 Coulter International Corp. Solid state RF oscillator-detector for flow cytometer
US6097485A (en) 1999-03-08 2000-08-01 Integrated Waveguides, Inc. Microchip optical transport technology for use in a personal flow cytometer
FR2791645B1 (en) 1999-04-02 2001-06-15 Valois Sa FLUID PRODUCT SAMPLE FOR PRESS
US6193647B1 (en) 1999-04-08 2001-02-27 The Board Of Trustees Of The University Of Illinois Microfluidic embryo and/or oocyte handling device and method
US6793387B1 (en) 1999-05-08 2004-09-21 Chata Biosystems, Inc. Apparatus for automatic preparation of a mixture and method
FR2793495B1 (en) 1999-05-14 2001-08-10 Imv Technologies THINNER FOR CRYOPRESERVATION OF CATTLE SPERMATOZOIDS
FR2793708B1 (en) 1999-05-21 2001-08-03 Valois Sa FLUID PRODUCT DISPENSING DEVICE
US6372506B1 (en) 1999-07-02 2002-04-16 Becton, Dickinson And Company Apparatus and method for verifying drop delay in a flow cytometer
IT1307787B1 (en) 1999-07-26 2001-11-19 Univ Firenze PROCESS TO INCREASE THE MOTILITY OF SPERMATOZOI AND SPERMATOZOIA SUPERIOR MOTILITY SO OBTAINED.
DE19935766A1 (en) * 1999-07-29 2001-02-01 Friedrich Schiller Uni Jena Bu Process for the optical excitation of fluorophore-labeled DNA and RNA
US6664550B2 (en) 1999-08-30 2003-12-16 Sandia National Laboratories Apparatus to collect, classify, concentrate, and characterize gas-borne particles
US6495366B1 (en) 1999-09-03 2002-12-17 Therakos, Inc. Uninterrupted flow pump apparatus and method
US6813017B1 (en) 1999-10-20 2004-11-02 Becton, Dickinson And Company Apparatus and method employing incoherent light emitting semiconductor devices as particle detection light sources in a flow cytometer
JP2003512616A (en) 1999-10-20 2003-04-02 ベクトン・ディキンソン・アンド・カンパニー Apparatus and method using incoherent light emitting semiconductor device as particle detection light source in flow cytometer
JP2003512605A (en) 1999-10-21 2003-04-02 サイトメーション, インコーポレイテッド Temporary dynamic flow cytometer analysis system
US6472153B1 (en) 1999-10-26 2002-10-29 Epoch Biosciences, Inc. Hybridization-triggered fluorescent detection of nucleic acids
US7208265B1 (en) 1999-11-24 2007-04-24 Xy, Inc. Method of cryopreserving selected sperm cells
JP5087192B2 (en) 1999-11-30 2012-11-28 インテレクソン コーポレイション Method and apparatus for selectively aiming a specific cell in a cell group
US6263745B1 (en) 1999-12-03 2001-07-24 Xy, Inc. Flow cytometer nozzle and flow cytometer sample handling methods
US6558911B1 (en) 1999-12-10 2003-05-06 Oregon Health Sciences University Sperm quality assay
ATE277566T1 (en) 2000-01-03 2004-10-15 Iberica De Reproduccion Asisti DEVICE FOR ARTIFICIAL FERTILIZATION OF PIGS
IT1317724B1 (en) 2000-01-14 2003-07-15 Istituto Sperimentale Italiano PROCEDURE FOR THE PRODUCTION OF NON-HUMAN SEX EMBRYOS WITH A HIGH GENETIC VALUE.
US6465169B2 (en) 2000-01-14 2002-10-15 Artemis Pharmaceuticals Gmbh Method for cryoconservation of Zebrafish sperm
EP1118268B1 (en) 2000-01-14 2002-04-17 ARTEMIS Pharmaceuticals GmbH Method for cryoconservation of zebrafish sperm
EP1257664A4 (en) 2000-01-28 2006-04-05 Althea Technologies Inc Methods for analysis of gene expression
US6587203B2 (en) 2000-02-17 2003-07-01 Cornell Research Foundation, Inc. Sort stream stabilizer for flow cytometer
US6618143B2 (en) 2000-02-18 2003-09-09 Idexx Laboratories, Inc. High numerical aperture flow cytometer and method of using same
US6646742B1 (en) 2000-02-19 2003-11-11 Mwi, Inc. Optical device and method for multi-angle laser light scatter
GB2360360B (en) 2000-03-14 2002-03-20 Univ Bristol A method of sorting cells
JP3587755B2 (en) 2000-03-14 2004-11-10 シスメックス株式会社 Particle measuring apparatus and method
US6482652B2 (en) 2000-03-23 2002-11-19 The Board Of Trustees Of The Leland Stanford Junior University Biological particle sorter
AU2001249589B2 (en) 2000-03-30 2006-07-13 Iowa State University Research Foundation Genetic markers for improved meat characteristics in animals
EP1290427B1 (en) 2000-04-11 2017-08-23 ChemoMetec A/S Method and apparatus for detecting fluorescence of a sample
EP1147774A1 (en) 2000-04-20 2001-10-24 Stichting Dienst Landbouwkundig Onderzoek Method for improving the quality of sperm for artificial insemination of animals
US7371517B2 (en) 2000-05-09 2008-05-13 Xy, Inc. High purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa
AU2001274864A1 (en) 2000-05-19 2001-12-03 Cytomation, Inc. A rapid multi-material sample input system
US6700130B2 (en) 2001-06-29 2004-03-02 Honeywell International Inc. Optical detection system for flow cytometry
US6590911B1 (en) 2000-06-02 2003-07-08 Coherent, Inc. Passively modelocked harmonic-generating laser
US6784981B1 (en) * 2000-06-02 2004-08-31 Idexx Laboratories, Inc. Flow cytometry-based hematology system
US7351376B1 (en) 2000-06-05 2008-04-01 California Institute Of Technology Integrated active flux microfluidic devices and methods
US6503698B1 (en) 2000-06-16 2003-01-07 The United States Of America As Represented By The Secretary Of Agriculture Cryopreservation of swine embryos
BR0111618A (en) 2000-06-12 2004-06-29 Xy Inc Integrated herd control system using isolated populations of x-chromosome-containing and y-chromosome-containing sperm
DE10031028B4 (en) 2000-06-26 2008-09-04 Gnothis Holding Sa Method for the selection of particles
GB0016920D0 (en) 2000-07-10 2000-08-30 Univ Cambridge Tech Decondensation of DNA
US20040005582A1 (en) * 2000-08-10 2004-01-08 Nanobiodynamics, Incorporated Biospecific desorption microflow systems and methods for studying biospecific interactions and their modulators
US20020131957A1 (en) 2000-08-10 2002-09-19 William Gavin Cryopreservation of sperm
FR2813283B1 (en) 2000-08-25 2003-02-14 Valois Sa INTEGRATED PUMP DISPENSER
EP1190684A1 (en) 2000-09-05 2002-03-27 Universiteit Gent Device and method for artificial insemination of bovines and other animals
US20020028434A1 (en) 2000-09-06 2002-03-07 Guava Technologies, Inc. Particle or cell analyzer and method
NZ524712A (en) 2000-09-08 2005-09-30 Univ Iowa State Res Found Inc Alternative forms of gene PRKAG3 as genetic markers associated with meat quality and reproductive traits in pigs
EP1334347A1 (en) 2000-09-15 2003-08-13 California Institute Of Technology Microfabricated crossflow devices and methods
GB0023041D0 (en) 2000-09-20 2000-11-01 Univ Manchester Identification apparatus
WO2002026114A2 (en) 2000-09-27 2002-04-04 Bitensky Mark W Cellular diagnostic arrays, methods of using and processes for producing same
CN1325909C (en) 2000-09-27 2007-07-11 清华大学 Apparatus for particle operation and guide and use method thereof
BR0005045A (en) 2000-09-28 2002-05-14 Marcos Fernando De Resende Mat Sperm sexing method using the classic complement system pathway, eliminating the alternative pathway by inactivating protein "b"
US7258774B2 (en) 2000-10-03 2007-08-21 California Institute Of Technology Microfluidic devices and methods of use
US20040031071A1 (en) * 2000-10-05 2004-02-12 Xy, Inc. System of hysteroscopic insemination of mares
AR029030A1 (en) 2000-10-05 2003-06-04 Xy Inc YEGUAS HYSTEROSCOPIC INSEMINATION PROVISION
WO2002031467A1 (en) 2000-10-12 2002-04-18 Amnis Corporation Multipass cavity for illumination and excitation of moving objects
JP2004537712A (en) 2000-10-18 2004-12-16 バーチャル・アレイズ・インコーポレーテッド Multiple cell analysis system
EP1328465A4 (en) 2000-10-23 2008-04-16 Medical Instill Tech Inc Fluid dispenser having a rigid vial and flexible inner bladder
AU2002220392A1 (en) 2000-11-09 2002-05-21 Universiy of Guelph Mammalian sex selection using genetic modification
SE0004777D0 (en) 2000-12-22 2000-12-22 Amersham Pharm Biotech Ab Separation of X and Y sperm cells
WO2002041906A2 (en) 2000-11-22 2002-05-30 Pharmacia Corporation Methods and apparatus for producing gender enriched sperm
CA2468772C (en) 2000-11-29 2013-10-29 George E. Seidel System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations
US6849423B2 (en) 2000-11-29 2005-02-01 Picoliter Inc Focused acoustics for detection and sorting of fluid volumes
US20020064809A1 (en) 2000-11-29 2002-05-30 Mutz Mitchell W. Focused acoustic ejection cell sorting system and method
US7713687B2 (en) * 2000-11-29 2010-05-11 Xy, Inc. System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations
US20020064808A1 (en) 2000-11-29 2002-05-30 Mutz Mitchell W. Focused acoustic energy for ejecting cells from a fluid
US20050048563A1 (en) 2000-11-30 2005-03-03 Novasite Pharmaceuticals, Inc Membrane molecule indicator compositions and methods
US6576291B2 (en) 2000-12-08 2003-06-10 Massachusetts Institute Of Technology Preparation of nanocrystallites
CA2440181A1 (en) 2000-12-15 2002-07-25 Dakocytomation Denmark A/S Electrical conductive containment system
US6577387B2 (en) 2000-12-29 2003-06-10 Johnson & Johnson Vision Care, Inc. Inspection of ophthalmic lenses using absorption
DE60221794D1 (en) 2001-01-29 2007-09-27 Univ Geneve PYRIMIDINIC AZYCLONUCLEOSIDES, PROCESS FOR THEIR PREPARATION AND THEIR USE
US6673095B2 (en) * 2001-02-12 2004-01-06 Wound Healing Of Oklahoma, Inc. Apparatus and method for delivery of laser light
US7029916B2 (en) 2001-02-21 2006-04-18 Maxcyte, Inc. Apparatus and method for flow electroporation of biological samples
US7015310B2 (en) 2001-03-12 2006-03-21 The State Of Oregon Acting By And Through The State Board Of Higher Education On Behalf Of The University Of Oregon Oxidation reduction sensitive green fluorescent protein variants
US6797139B2 (en) * 2001-03-19 2004-09-28 Applera Corporation Detection cell for guiding excitation light therein and method for using same
US6706163B2 (en) 2001-03-21 2004-03-16 Michael Seul On-chip analysis of particles and fractionation of particle mixtures using light-controlled electrokinetic assembly of particles near surfaces
CA2442019A1 (en) 2001-03-22 2002-10-03 Infigen, Inc. Sex-specific selection of sperm from transgenic animals
EP1565747B1 (en) 2001-03-29 2013-11-27 Cellect Technologies Corp. Method and system for separating and sorting particles
EP1245944B1 (en) 2001-03-29 2007-02-14 Sysmex Corporation Flow cytometer
EP1377811B1 (en) 2001-04-03 2008-07-16 Micronics, Inc. Split focusing cytometer
JP2002311027A (en) 2001-04-09 2002-10-23 Hitachi Software Eng Co Ltd Beads, manufacturing method of beads, flow cytometer, and program
US6416190B1 (en) 2001-04-27 2002-07-09 University Of Chicago Apparatus for using optical tweezers to manipulate materials
US20020186375A1 (en) 2001-05-01 2002-12-12 Asbury Charles L. Device and methods for detecting samples in a flow cytometer independent of variations in fluorescence polarization
WO2002092161A1 (en) 2001-05-10 2002-11-21 Biophan, Llc Miniaturized particle analyzer
US7345758B2 (en) 2001-05-17 2008-03-18 Cytopeia Apparatus for analyzing and sorting biological particles
EP1395374B1 (en) 2001-05-17 2013-04-17 Beckman Coulter, Inc. Flow cytometer with active automated optical alignment system
US20020182590A1 (en) 2001-05-25 2002-12-05 Vanderbilt University Determining protein function in cell culture using RNA interference
US20030048433A1 (en) 2001-06-01 2003-03-13 Jean-Marie Desjonqueres Cytometer signal processing system and method
FR2825987B1 (en) 2001-06-19 2003-12-12 Valois Sa FLUID PRODUCT DISPENSER
US7105355B2 (en) 2001-07-18 2006-09-12 The Regents Of The University Of Michigan Flow cytometers and detection system of lesser size
US7704728B2 (en) 2001-07-18 2010-04-27 The University Of Michigan Microfluidic gravity pump with constant flow rate
FR2828281B1 (en) 2001-08-02 2004-12-31 Biocytex DEVICE FOR ANALYZING A SAMPLE IN PARTICULAR BY FLOW CYTOMETRY
WO2003020877A2 (en) 2001-09-01 2003-03-13 Pharmacia Corporation Staining and sorting genomes and chromosomes using sequence-specific polyamides
DE10151216A1 (en) 2001-10-16 2003-04-24 Zeiss Carl Jena Gmbh Method for the optical detection of characteristic quantities of an illuminated sample
US20030078703A1 (en) 2001-10-19 2003-04-24 Surromed, Inc. Cytometry analysis system and method using database-driven network of cytometers
US6838289B2 (en) 2001-11-14 2005-01-04 Beckman Coulter, Inc. Analyte detection system
US6831279B2 (en) 2001-11-27 2004-12-14 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of National Defence Laser diode-excited biological particle detection system
AU2002361220A1 (en) 2001-12-31 2003-07-15 Institut Fur Physikalische Hochtechnologie E.V. Micro-recess array for size-dependent sorting or separation of cells suspended in a flowing medium and method for analysing the functional activity of individual cells
WO2003056330A2 (en) 2001-12-31 2003-07-10 Institut für Physikalische Hochtechnologie e.V. Cell sorting system for the size-based sorting or separation of cells suspended in a flowing fluid
US6780377B2 (en) 2002-01-22 2004-08-24 Dakocytomation Denmark A/S Environmental containment system for a flow cytometer
US6698627B2 (en) 2002-02-19 2004-03-02 Valois S.A.S. Fluid dispenser
US6849394B2 (en) 2002-02-21 2005-02-01 Minitube Of America Compositions comprising reproductive cell media and methods for using such compositions
AU2003219836A1 (en) 2002-02-27 2003-09-09 The Regents Of The University Of Michigan Process for sorting motile particles from lesser-motile particles and apparatus suitable therefor
US20030175980A1 (en) 2002-03-14 2003-09-18 Hayenga Jon W. Ribbon flow cytometry and cell sorting
US7223371B2 (en) 2002-03-14 2007-05-29 Micronics, Inc. Microfluidic channel network device
JP2004000144A (en) 2002-03-29 2004-01-08 Aisin Seiki Co Ltd Cell separating and sorting apparatus and substrate for cell alignment
US7312085B2 (en) * 2002-04-01 2007-12-25 Fluidigm Corporation Microfluidic particle-analysis systems
EP1532563A2 (en) 2002-06-28 2005-05-25 Monsanto Technology LLC, Patent Departement Swine genetics business system
AU2003255529B2 (en) 2002-07-10 2008-11-20 Laboratoires Serono Sa Use of compounds for increasing spermatozoa motility
BR0312843A (en) 2002-07-22 2005-05-10 Xy Inc System to process sperm cells
DK2275533T3 (en) 2002-08-01 2016-11-07 Xy Llc Sperm Cell Assessment Method
AU2003265471B2 (en) 2002-08-15 2009-08-06 Xy, Llc. High resolution flow cytometer
US7169548B2 (en) * 2002-09-13 2007-01-30 Xy, Inc. Sperm cell processing and preservation systems
US20040061853A1 (en) 2002-09-27 2004-04-01 Blasenheim Barry J. Prism-based flow cytometry excitation optics
US7201875B2 (en) * 2002-09-27 2007-04-10 Becton Dickinson And Company Fixed mounted sorting cuvette with user replaceable nozzle
US6941005B2 (en) 2002-11-01 2005-09-06 Coulter International Corp. Monitoring and control of droplet sorting
AU2003298682A1 (en) 2002-11-20 2004-06-15 University Of Virginia Patent Foundation Isolation of sperm cells from other biological materials using microfabricated devices and related methods thereof
AU2003297304A1 (en) 2002-12-19 2004-07-22 Monsanto Technology Llc Method and means for early detection of pregnancy in animals by combination testing
EP2309245B1 (en) 2003-03-28 2015-11-25 Inguran, LLC Methods for providing sex-sorted animal sperm
US7335507B2 (en) 2003-03-28 2008-02-26 Monsanto Technology Llc Process for the staining of sperm
US20060263829A1 (en) 2003-05-15 2006-11-23 Evans Kenneth M Efficient haploid cell sorting flow cytometer systems
US20050011582A1 (en) 2003-06-06 2005-01-20 Haug Jeffrey S. Fluid delivery system for a flow cytometer
US7460223B2 (en) * 2003-09-19 2008-12-02 Applied Biosystems Inc. Inverted orientation for a microplate
EP1730523B1 (en) 2004-03-29 2010-01-13 Inguran, LLC Use of a composition which regulates oxidation/reduction reactions intracellularly and/or extracellularly in a staining or sorting process of spermatozoa
CA2561519C (en) 2004-03-29 2017-08-22 Monsanto Technology Llc Sperm dispersions for use in insemination
EP2269617B1 (en) 2004-07-22 2016-04-27 Inguran, LLC Process for enriching a population of sperm cells
PT2884258T (en) 2004-07-27 2016-12-13 Beckman Coulter Inc Enhancing flow cytometry discrimination with computer-implemented geometric transformation
US20060118167A1 (en) 2004-12-03 2006-06-08 Xy, Inc. Pressure regulated continuously variable volume container for fluid delivery
US20060147894A1 (en) 2004-12-30 2006-07-06 Vicam, L.P. Jacketed vessel for holding semen for sex biasing mammals through artificial insemination and systems and methods for enhancing the probability of sex biasing using the same
US7618770B2 (en) 2005-07-29 2009-11-17 Xy, Inc. Methods and apparatus for reducing protein content in sperm cell extenders
CA2626518A1 (en) * 2005-10-19 2007-04-26 Scott Josephson Improved reproductive management
DE202006018701U1 (en) 2006-12-05 2007-03-01 Takata-Petri Ag Housing for air bag arrangement of passenger safety device, has two housing parts which have side wall locking with connecting edge whereby housing parts with closed housing lie on each other over connecting edge
CN100998524A (en) 2007-01-19 2007-07-18 北京锦绣大地农业股份有限公司 Method for generating sex controlled ox-embryo
PL2304020T3 (en) * 2008-06-30 2019-06-28 Microbix Biosystems Inc. Method and apparatus for sorting cells
DE102013226771A1 (en) 2013-12-19 2015-06-25 Robert Bosch Gmbh Method and device for mass decoupling for a motor vehicle

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4179218A (en) * 1978-05-15 1979-12-18 The Boeing Company Particle size analyzer
US4274740A (en) * 1978-07-21 1981-06-23 Eidenschink Herrn H Optical process and apparatus for determining particle size of colloidal solution
US5132548A (en) * 1990-09-14 1992-07-21 High Yield Technology High sensitivity, large detection area particle sensor for vacuum applications
US5471299A (en) * 1992-02-21 1995-11-28 The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland Apparatus and method for the analysis of particle characteristics using monotonically scattered light

Cited By (50)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7929137B2 (en) 1997-01-31 2011-04-19 Xy, Llc Optical apparatus
US6372422B1 (en) 1997-12-31 2002-04-16 Colorado State University Through Its Agent Colorado State University Research Foundation And Xy, Inc. Multiple sexed embryo production system for mammals
US6524860B1 (en) 1997-12-31 2003-02-25 Xy, Inc. Methods for improving sheath fluids and collection systems for sex-specific cytometer sorting of sperm
US9422523B2 (en) 1997-12-31 2016-08-23 Xy, Llc System and method for sorting cells
US9365822B2 (en) 1997-12-31 2016-06-14 Xy, Llc System and method for sorting cells
US7820425B2 (en) 1999-11-24 2010-10-26 Xy, Llc Method of cryopreserving selected sperm cells
US10208345B2 (en) 2000-05-09 2019-02-19 Xy, Llc Method for producing high purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa
JP2012005490A (en) * 2000-05-09 2012-01-12 Xy Llc High purity x-chromosome bearing and y-chromosome bearing populations of spermatozoa
EP2258172B1 (en) * 2000-05-09 2017-04-19 Xy, Llc Flow cytometer for diffentiating x-chromosome bearing and y-chromosome bearing populations of spermatozoa
US9145590B2 (en) 2000-05-09 2015-09-29 Xy, Llc Methods and apparatus for high purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa
WO2002028311A1 (en) 2000-10-05 2002-04-11 Xy, Inc. System of hysteroscopic insemination of mares
US8652769B2 (en) 2000-11-29 2014-02-18 Xy, Llc Methods for separating frozen-thawed spermatozoa into X-chromosome bearing and Y-chromosome bearing populations
US7771921B2 (en) 2000-11-29 2010-08-10 Xy, Llc Separation systems of frozen-thawed spermatozoa into X-chromosome bearing and Y-chromosome bearing populations
US7713687B2 (en) 2000-11-29 2010-05-11 Xy, Inc. System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations
US9879221B2 (en) 2000-11-29 2018-01-30 Xy, Llc Method of in-vitro fertilization with spermatozoa separated into X-chromosome and Y-chromosome bearing populations
EP1395374B1 (en) * 2001-05-17 2013-04-17 Beckman Coulter, Inc. Flow cytometer with active automated optical alignment system
EP1395374A1 (en) * 2001-05-17 2004-03-10 Cytomation, Inc. Flow cytometer with active automated optical alignment system
US8497063B2 (en) 2002-08-01 2013-07-30 Xy, Llc Sex selected equine embryo production system
US8486618B2 (en) 2002-08-01 2013-07-16 Xy, Llc Heterogeneous inseminate system
WO2004012837A2 (en) 2002-08-01 2004-02-12 Xy, Inc. Low pressure sperm cell separation system
US6936828B2 (en) 2003-02-14 2005-08-30 Honeywell International Inc. Particle detection system and method
WO2004074817A1 (en) * 2003-02-14 2004-09-02 Honeywell International Inc. Particle detection system and method
US11104880B2 (en) 2003-03-28 2021-08-31 Inguran, Llc Photo-damage system for sorting particles
US8748183B2 (en) 2003-03-28 2014-06-10 Inguran, Llc Method and apparatus for calibrating a flow cytometer
US10100278B2 (en) 2003-03-28 2018-10-16 Inguran, Llc Multi-channel system and methods for sorting particles
US9377390B2 (en) 2003-03-28 2016-06-28 Inguran, Llc Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm
US7943384B2 (en) 2003-03-28 2011-05-17 Inguran Llc Apparatus and methods for sorting particles
US7799569B2 (en) 2003-03-28 2010-09-21 Inguran, Llc Process for evaluating staining conditions of cells for sorting
US11718826B2 (en) 2003-03-28 2023-08-08 Inguran, Llc System and method for sorting particles
US8664006B2 (en) 2003-03-28 2014-03-04 Inguran, Llc Flow cytometer apparatus and method
US7758811B2 (en) 2003-03-28 2010-07-20 Inguran, Llc System for analyzing particles using multiple flow cytometry units
US8709825B2 (en) 2003-03-28 2014-04-29 Inguran, Llc Flow cytometer method and apparatus
US8709817B2 (en) 2003-03-28 2014-04-29 Inguran, Llc Systems and methods for sorting particles
US9040304B2 (en) 2003-03-28 2015-05-26 Inguran, Llc Multi-channel system and methods for sorting particles
US7723116B2 (en) 2003-05-15 2010-05-25 Xy, Inc. Apparatus, methods and processes for sorting particles and for providing sex-sorted animal sperm
US7892725B2 (en) 2004-03-29 2011-02-22 Inguran, Llc Process for storing a sperm dispersion
US7838210B2 (en) 2004-03-29 2010-11-23 Inguran, LLC. Sperm suspensions for sorting into X or Y chromosome-bearing enriched populations
US7833147B2 (en) 2004-07-22 2010-11-16 Inguran, LLC. Process for enriching a population of sperm cells
KR101170859B1 (en) 2004-07-30 2012-08-02 바이오비질런트 시스템즈 인코포레이티드 Pathogen and particle detector system and method
WO2006073492A3 (en) * 2004-07-30 2007-07-05 Biovigilant Systems Inc Pathogen and particle detector system and method
US9013692B2 (en) 2008-06-12 2015-04-21 East Carolina University Flow cytometer apparatus for three dimensional difraction imaging and related methods
US8467040B2 (en) 2008-06-30 2013-06-18 Microbix Biosystems, Inc. Method and apparatus for sorting cells
US8004661B2 (en) 2008-06-30 2011-08-23 Microbix Biosystems Inc. Method and apparatus for sorting cells
US8634077B2 (en) 2008-10-01 2014-01-21 East Carolina University Methods and systems for optically characterizing a turbid material using a structured incident beam
US8665439B2 (en) 2009-06-30 2014-03-04 Microbix Biosystems, Inc. Method and apparatus for limiting effects of refraction in cytometry
US9556416B2 (en) 2011-02-15 2017-01-31 Microbix Biosystems Inc. Methods, systems and apparatus for performing flow cytometry
US10465163B2 (en) 2011-02-15 2019-11-05 Microbix Biosystems Inc. Methods, systems, and apparatus for performing flow cytometry
US8919194B2 (en) 2012-09-05 2014-12-30 Simmonds Precision Products, Inc. Liquid level gauging system with bubble shedding reflectors
EP2720007A1 (en) * 2012-09-05 2014-04-16 Simmonds Precision Products, Inc. Liquid level gauging system with bubble shedding reflectors
US10101274B2 (en) 2012-12-05 2018-10-16 Genepoc Inc. Optical interrogation device

Also Published As

Publication number Publication date
US7586604B2 (en) 2009-09-08
EP2264428A3 (en) 2014-05-21
CA2279574A1 (en) 1998-08-06
JP6016830B2 (en) 2016-10-26
JP2011180144A (en) 2011-09-15
US20150204772A1 (en) 2015-07-23
US20070285662A1 (en) 2007-12-13
EP2264428B1 (en) 2017-05-03
JP5512579B2 (en) 2014-06-04
JP2001509266A (en) 2001-07-10
EP1017987A1 (en) 2000-07-12
US20050110996A1 (en) 2005-05-26
US8553226B2 (en) 2013-10-08
JP2007183655A (en) 2007-07-19
EP1562035A3 (en) 2014-08-20
DE69830598D1 (en) 2005-07-21
US20110195492A1 (en) 2011-08-11
ATE298084T1 (en) 2005-07-15
JP4323571B2 (en) 2009-09-02
DE69830598T2 (en) 2006-05-18
US6819411B1 (en) 2004-11-16
EP1017987A4 (en) 2002-08-21
AU752985B2 (en) 2002-10-03
EP2264427A2 (en) 2010-12-22
US7221453B2 (en) 2007-05-22
AU5783698A (en) 1998-08-25
EP1017987B1 (en) 2005-06-15
EP2264428A2 (en) 2010-12-22
US7929137B2 (en) 2011-04-19
JP2014112108A (en) 2014-06-19
US20100007880A1 (en) 2010-01-14
EP2264427A3 (en) 2014-05-21
EP2264427B1 (en) 2017-05-03
CA2279574C (en) 2007-07-24
US8975035B2 (en) 2015-03-10
US20130162993A1 (en) 2013-06-27
US8610892B2 (en) 2013-12-17
EP1562035B1 (en) 2017-01-25
EP1562035A2 (en) 2005-08-10
US20110071035A1 (en) 2011-03-24
JP4727595B2 (en) 2011-07-20

Similar Documents

Publication Publication Date Title
US6819411B1 (en) Optical apparatus
US7075647B2 (en) Back-scatter detection in flow cytometers
US4281924A (en) Reflector for the laser beam of a particle analyzer
Sharpe et al. New optical configuration for flow cytometric sorting of aspherical cells
AU2008200836B2 (en) Optical Apparatus
CA2588622C (en) Optical apparatus
AU2002318853B2 (en) Optical Apparatus
AU2005203372B2 (en) Optical Apparatus
NZ523347A (en) Optical apparatus with annular beam and focussing reflector for sperm particle irradiance analysis
NZ522984A (en) Optical flow cytometry
JPS5935130A (en) Flow-cell
NZ337340A (en) Flow cytometer inspection light radially symmetrically converged on linear particle stream

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GE GH GM GW HU ID IL IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
ENP Entry into the national phase

Ref document number: 2279574

Country of ref document: CA

Ref country code: CA

Ref document number: 2279574

Kind code of ref document: A

Format of ref document f/p: F

Ref country code: JP

Ref document number: 1998 532763

Kind code of ref document: A

Format of ref document f/p: F

WWE Wipo information: entry into national phase

Ref document number: 337340

Country of ref document: NZ

WWE Wipo information: entry into national phase

Ref document number: 57836/98

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 1998901601

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 09355461

Country of ref document: US

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWP Wipo information: published in national office

Ref document number: 1998901601

Country of ref document: EP

WWG Wipo information: grant in national office

Ref document number: 57836/98

Country of ref document: AU

WWG Wipo information: grant in national office

Ref document number: 1998901601

Country of ref document: EP