WO2006026766A2 - Frequency-multiplexed detection of multiple wavelength light for flow cytometry - Google Patents
Frequency-multiplexed detection of multiple wavelength light for flow cytometry Download PDFInfo
- Publication number
- WO2006026766A2 WO2006026766A2 PCT/US2005/031493 US2005031493W WO2006026766A2 WO 2006026766 A2 WO2006026766 A2 WO 2006026766A2 US 2005031493 W US2005031493 W US 2005031493W WO 2006026766 A2 WO2006026766 A2 WO 2006026766A2
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- WIPO (PCT)
- Prior art keywords
- light
- detector
- wavelength
- frequency
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1456—Electro-optical investigation, e.g. flow cytometers without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
- G01N15/1459—Electro-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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
- G01N2015/1438—Using two lasers in succession
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N2015/1477—Multiparameters
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N2015/1486—Counting the particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/06—Illumination; Optics
- G01N2201/069—Supply of sources
- G01N2201/0691—Modulated (not pulsed supply)
Definitions
- This invention pertains to cytometers and particularly to optical systems of cytometers. More particularly, the invention pertains to the optical acquisition of information about microscopic particles or components in a flow stream of a cytometer.
- This invention is related to U.S. Patent Application Serial No. 10/225,325, by Bernard Fritz et al . , filed August 21, 2002, and entitled “Optical Alignment Detection System” , which is incorporated herein by reference; and the invention is related to U.S. Patent Application Serial No. 10/304,773, to Aravind Padmanabhan et al., filed November 26, 2002, and entitled “Portable Scattering and Fluorescence Cytometer” , which is incorporated herein by reference.
- This invention also is related to U.S. Patent No. 6,549,275 Bl, by Cabuz et al . , issued April 15, 2003, and entitled "Optical Detection
- fluid may be used herein as a generic term that includes gases and liquids as species. For instance, air, gas, water and oil are fluids.
- the invention is an optical system for a cytometer using a multiplexing scheme to detect light of various wavelengths to obtain information relative to the particles that the light is impinging in a flow channel of the cytometer.
- Figure 1 shows a multiplexed multiple wavelength light scattering system with a single detector
- Figure 2 is a graph of light signals versus their respective modulation frequencies.
- Figure 3 is a diagram of a cytometer as an illustrative example that may use the multiplexed multiple wavelength light scattering system.
- Improved performance may be achieved by measuring optical scattering properties of a particle at multiple wavelengths.
- the invention may provide a way to accomplish this measuring approach by using a single detector assembly for all wavelengths.
- Each wavelength light source may be modulated at a unique frequency sufficiently separated from the other modulated sources to enable its signal to be demultiplexed unambiguously at the output of the detector.
- Light from all modulated sources scattered by the particle under measurement may be collected on the same detector assembly.
- improved differentiation and accuracy in counting and distinguishing multiple particle types may be achieved by performing multi-dimensional measurements, such as particle volume, scattering at various angles, and scattering in various wavelengths.
- the invention may reveal improvements to this optical interrogation technique (i.e., multi-wave scattering) .
- Scattering at multiple wavelengths may be done at spatially separated locations along the flow channel. This may require careful synchronization in timing as well as multiple detector arrays and spectra filters.
- This difficulty may be avoided by the use of modulation frequency multiplexing of the various wavelength sources.
- Each source may be modulated at a unique and sufficiently high frequency to meet system bandwidth requirements.
- the sources may be folded into one optical input path and focused simultaneously onto the same particle location.
- FIG. 1 shows an illustrative example implementing the invention. This figure shows a cross-section view of a channel 11.
- Channel 11 may be a flow or measurement channel of a cytometer. It may have a core stream having particles 12 moving through channel 11. The core stream with particles 12 may be looked at as flowing into the surface of the figure. Channel 11 may be lengthy. The core stream along with particles 12 may be kept away from the inside surfaces of channel 11 with a sheathing fluid that surrounds the core stream.
- the location of the cross-section of channel 11 may be where a light source and detector arrangement may be placed.
- Channel 11 may have transparent windows 13 and 14 to facilitate the light source detector arrangement.
- a light beam 15 may enter channel 11 through window 13, impinge a particle 12 which may scatter beam 15 into light 16 which may exit channel 11 through window 14.
- Light 16 may be sensed by a detector 17.
- Detector 17 may be an annular type having a ring of surface area 18 sensitive to light.
- the detector 17 may be expanded with another ring of surface area 19 also sensitive to light 16.
- Light sensitive surfaces 18 and 19 may be isolated form each other by an annular area 21 that is not sensitive to light.
- detector 17 may be further expanded with a central light-sensitive area 22 that may be isolated from the light-sensitive annular area 19 by an annular area 23 that is not sensitive to light.
- the detector 17 may be expanded to include as many annular detectors, each subtending its own prescribed angular interval, as needed.
- the annular detectors or other kinds of detectors of an array of the detector may provide electrical signals representing light impinging the detector at respective angles. That is, one electrical signal may represent detected light of a first angle; another electrical signal may represent detected light of a second angle; and so on.
- Various kinds of information may be obtained about the particles 12 from the scattered light. First, a count of the particles 12 may be made with the successive interruption of the light beam 15 to detector 17.
- particles 12 may be obtained from scattered light that impinges detector 17.
- the magnitudes of the scattered light and the location of such light on detector 17 may be noted electronically from the signals from the various detector 17 surfaces.
- Another dimension of information may be obtained from the scattered light if the various wavelengths of the scattered light are known.
- Light 15 beams of various wavelengths may scatter differently from particles 12. That is, a light beam of one wavelength may scatter differently than a light beam of another wavelength for the same point of impingement of a particle, or even the same particle, in the same location. These differences of scattering may provide additional information about the particle.
- Beam 15 may be composed of light from a number (n) of light sources 24, 25 and 26.
- Light source 24 may emit or emanate a light beam 27 having a wavelength /L 1 .
- Light source 25 may
- light source 25 and light source 26 may be numerous similar light sources with light beams having different wavelengths, respectively.
- Beam 27 may propagate from source 24 to a component dichroic mirror 31 in a dichroic fold mirror assembly 30.
- Mirror 31 may reflect at least a portion of beam 27 approximately 90 degrees towards channel 11.
- Beam 28 may propagate to a dichroic mirror 32 of assembly 30.
- Mirror 32 may deflect and/or reflect at least a portion of beam 28 approximately 90 degrees towards channel 11.
- Beam 29 may propagate to a dichroic mirror 33 of assembly 30.
- Mirror 33 may reflect at least a portion of beam 29 approximately 90 degrees towards channel 11.
- beam 28 may, at least in part, go through mirror 33 and any mirrors between mirrors 32 and 33.
- a resultant beam 15, which may include beams 27, 28 and 29 and any beams reflected or deflected by other mirrors situated between mirrors 32 and 33 of assembly 30. Beam 15 may proceed through aperture 34, optics 35 and window 13 of channel 11. Since beam 15 may go through window 13 of channel
- a modulator 36 may modulate the output of the light source 24 with a frequency fi.
- a modulator 37 may modulate the output of light source 25 with a frequency f 2 and modulator 38 may modulate the output of light source 26 with a frequency f n .
- modulators 37 and 38 there may be other modulators that modulate additional light sources of other wavelengths that may be situated between light sources 25 and 26. This approach may be regarded as a frequency multiplexing of the light sources.
- Modulators 36, 37, 38 and the other modulators may be connected to and controlled by computer/processor 40.
- the output of detector 17 may go to a frequency analyzer 39 which may demultiplex the detected light 16 and 15 signals and separate out the light into component signals according to their wavelengths and respective light sources. These signals may be provided to the computer/processor 40 for analysis, counting, identification, recording and/or other actions.
- Modulation frequencies may be relatively high in comparison to signal frequencies.
- Figure 2 reveals a graph of the signals multiplexed according to frequency.
- a signal 41 may be of the wavelength A 1 multiplexed at 10.0 MHz
- a signal 42 may be of the wavelength ⁇ 2 multiplexed at 10.3 MHz
- a signal 41 may be of the wavelength A 1 multiplexed at 10.0 MHz
- a signal 42 may be of the wavelength ⁇ 2 multiplexed at 10.3 MHz
- a signal 41 may be of the wavelength A 1 multiplexed at 10.0 MHz
- a signal 42 may be of the wavelength ⁇ 2 multiplexed at 10.3 MHz
- a signal 41 may be of the wavelength A 1 multiplexed at 10.0 MHz
- a signal 42 may be of the wavelength ⁇ 2 multiplexed at 10.3 MHz
- a signal 41 may be of the wavelength A 1 multiplexed at 10.0 MHz
- a signal 42 may be of the wavelength ⁇ 2 multiplexed at
- signal 43 may be of the wavelength ⁇ n multiplexed at 10.6 MHz. Additional signals of other wavelengths may be multiplexed at other frequencies for demultiplexing at the output of the detector 17.
- FIG. 3 is a diagram of a cytometer 45 that may incorporate an illustrative application of the multiplexed multiple wavelength light scattering system.
- Cytometer 45 may have a channel 11 with a core stream of particles 12.
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2007530436A JP2008511843A (en) | 2004-09-01 | 2005-09-01 | Frequency multiplexed detection of multiwavelength light for flow cytometry. |
CN2005800366981A CN101052867B (en) | 2004-09-01 | 2005-09-01 | Frequency-multiplexed detection of multiple wavelength light for flow cytometry |
EP05794052.0A EP1784630B1 (en) | 2004-09-01 | 2005-09-01 | Frequency-multiplexed detection of multiple wavelength light for flow cytometry |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/931,686 US7612871B2 (en) | 2004-09-01 | 2004-09-01 | Frequency-multiplexed detection of multiple wavelength light for flow cytometry |
US10/931,686 | 2004-09-01 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2006026766A2 true WO2006026766A2 (en) | 2006-03-09 |
WO2006026766A3 WO2006026766A3 (en) | 2006-06-01 |
Family
ID=35429242
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2005/031493 WO2006026766A2 (en) | 2004-09-01 | 2005-09-01 | Frequency-multiplexed detection of multiple wavelength light for flow cytometry |
Country Status (5)
Country | Link |
---|---|
US (1) | US7612871B2 (en) |
EP (1) | EP1784630B1 (en) |
JP (2) | JP2008511843A (en) |
CN (1) | CN101052867B (en) |
WO (1) | WO2006026766A2 (en) |
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US9260693B2 (en) | 2004-12-03 | 2016-02-16 | Cytonome/St, Llc | Actuation of parallel microfluidic arrays |
KR20070104347A (en) | 2004-12-03 | 2007-10-25 | 사이토놈, 인크. | Unitary cartridge for particle processing |
WO2012036794A2 (en) * | 2010-08-01 | 2012-03-22 | Array Optronix, Inc. | Photodetectors with light incident surface and contact surface and applications thereof |
CN102288531B (en) * | 2011-04-21 | 2012-12-19 | 西北工业大学 | Method and device for filtering, dyeing and counting cells |
EP3521810B1 (en) * | 2018-01-31 | 2019-11-27 | SICK Engineering GmbH | Analyser for the determination of fine dust |
CN114152559A (en) * | 2021-12-13 | 2022-03-08 | 常州必达科生物科技有限公司 | Light spot detection method and device |
CN114324205A (en) * | 2021-12-16 | 2022-04-12 | 赛默飞世尔(上海)仪器有限公司 | Method and system for performing spectrometry on sample and flow cytometer |
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JP2008511843A (en) | 2008-04-17 |
JP2011059127A (en) | 2011-03-24 |
EP1784630B1 (en) | 2018-05-30 |
EP1784630A2 (en) | 2007-05-16 |
WO2006026766A3 (en) | 2006-06-01 |
JP5762720B2 (en) | 2015-08-12 |
US7612871B2 (en) | 2009-11-03 |
US20060051096A1 (en) | 2006-03-09 |
CN101052867A (en) | 2007-10-10 |
CN101052867B (en) | 2012-05-02 |
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