WO1997022870A1 - Multi-parameter hematology apparatus and method - Google Patents
Multi-parameter hematology apparatus and method Download PDFInfo
- Publication number
- WO1997022870A1 WO1997022870A1 PCT/US1996/019823 US9619823W WO9722870A1 WO 1997022870 A1 WO1997022870 A1 WO 1997022870A1 US 9619823 W US9619823 W US 9619823W WO 9722870 A1 WO9722870 A1 WO 9722870A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- blood
- light
- rbc
- ensemble
- wbc
- Prior art date
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5094—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for blood cell populations
-
- 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
- G01N15/02—Investigating particle size or size distribution
- G01N15/0205—Investigating particle size or size distribution by optical means, e.g. by light scattering, diffraction, holography or imaging
- G01N15/0211—Investigating a scatter or diffraction pattern
-
- 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
- G01N15/02—Investigating particle size or size distribution
- G01N15/0205—Investigating particle size or size distribution by optical means, e.g. by light scattering, diffraction, holography or imaging
- G01N15/0227—Investigating particle size or size distribution by optical means, e.g. by light scattering, diffraction, holography or imaging using imaging, e.g. a projected image of suspension; using holography
Definitions
- This invention relates to diagnostic hematology apparatus and, more particularly, to apparatus and methods for obtaining diagnostic information such as cell concentration and sizing using ensemble light scattering.
- One means of estimating the concentration of blood cells without dilution is by packing the cells with centrifugal force and measuring the length of the packed column relative to the total length. Since the cells of different class (RBC WBC and PLT) have different density, they may be layered by this procedure allowing a measurement of concentration by class.
- This general method has been developed for use in remote areas such as those listed above. Becton Dickenson has marketed a device based on a these principles called the QBC. However, this device has two serious deficiencies, i.e., accurate operation requires operator skill in visually estimating the boundaries between cell classes; and the device is unable to determine the mean red blood cell volume which is an important measurement in the diagnosis of anemia.
- Another means of estimating the concentration and/or size of cells at low dilution is to observe the diffraction and/or scattering of light by ensembles of blood cells.
- This method has been used to estimate cell concentration by measuring the turbidity of cell suspension, to estimate cell size by measuring the diameter of the rings formed when light is diffracted by RBC dried on a glass slide or in liquid suspension, and to discriminate between cell population based on the angular distribution of the scattered light and to measure the average deformation of RBC when under hydrodynamic stress.
- Ensemble light scattering is used as the fundamental technology for a multi-parameter hematology analyzer. It is expected by considering the application of this technology, benefit may be gained by synergy with advances in related technology which make the application more practical today. Such advances include the common availability (at low cost) of coherent light sources (lasers) and the growing availability (also at low cost) of the software and hardware required to digitize and analyze electronic images. Such devices as CCD cameras, personal computers and video phone interphases developed for the consumer market may be applicable directly to this device.
- a third benefit is derived in that the measurement time is reduced.
- Instruments which count cells one at a time must allow time for the accumulation of cell counts, whereas the combined effect of a multiplicity of cells are measured instantaneously when ensemble scattering is employed.
- a fourth benefit may result from applying the same root technology to other aspects of diagnostic hematology. For instance, ensemble scattering has been used to measure RBC deformability and aggregation of platelets. Ensemble scattering may also be useful in measuring the cell/cell interaction which are the dominant factor in the sedimentation of erythrocytes. Thus, these additional measurements may be more easily combined into a single device which in addition to the traditional measurement of cell count and cell size also measures the ESR.
- FIG. 1 is a block diagram of an apparatus in accordance with the present invention.
- the scattering of coherent light by an ensemble of particles which all are within the coherence length of light source is obtained by the superposition of the light scattered by individual particles.
- the net scattering distribution is N times the scattering intensity of a single particle where N is the number of particles. This is also a well known effect which has been used to both size and estimate the concentration of particles.
- the tendency of particles to aggregate may be determined by measuring the fluctuations in intensity of the
- ° Hematology parameters discernable using ensemble light scattering include: white blood cell count (WBC); red blood cell count (RBC); platelet count (PLT); mean cell volume (MCV); mean cell hemoglobin (MCH); mean cell hemoglobin concentration (MCHC); percentage of granulocytes (GRAN); percentage 5 of agranulocytes (AGRAN); percentage of polymorphonuclear cells (POLYS); percentage of mononuclear cells (MONO); and erythrocyte sedimentation rate (ESR).
- WBC white blood cell count
- RBC red blood cell count
- PHT platelet count
- MCV mean cell volume
- MH mean cell hemoglobin
- MCHC mean cell hemoglobin concentration
- GRAN percentage of granulocytes
- AGRAN percentage 5 of agranulocytes
- POLYS percentage of polymorphonuclear cells
- MONO percentage of mononuclear cells
- ESR erythrocyte sedimentation rate
- FIG. 1 illustrates a specific example of the concept of using ensemble scattering to obtain the measurement of RBC.
- the light from a laser is scattered by a thin transparent specimen container containing a low (10-100) dilution of whole blood.
- Ne, Gas, GAAS diode lasers The specifications for the preferred laser are that it emits in the red spectrum and is of a beam diameter which is less than the specimen container width.
- the coherence should be greater than cell length and the power level is preferably about 1 milliwatt.
- the specimen container should have a high degree of optical clarity and a path length through the container of at least about 0.1mm.
- the optical path length should be such that there is a reasonable probability that the light will be scattered by one blood cell while a much lower probability that the light will be multiply scattered by several blood cells. For this reason the optical path length should be roughly equal to the average distance between blood cells. For example:
- Such a specimen container is the BMS-10150, available from Federich Dimmock Co.
- the BMS-10150 has dimensions of 0.1mm x 1.0mm x 50mm.
- the pattern from the scattering is imaged on a CCD camera.
- CCD camera is preferably monochromatic with at least 350,000 pixel capacity.
- One such camera is the model KRMI, available from Hitachi Denshi, Ltd.
- the resultant image is then captured and digitized through a digitizing means.
- the digitizing means is also preferably monochromatic with at least 8 bit high accuracy analog to digital capability.
- a digitizing means meeting these requirements is the model DT 3155 Frame Grabber, available from Data Translation.
- the digitized image is then analyzed with a small computer containing a resident image processing software program.
- the computer can be any of a wide variety of known microprocessors including the 486 and Pentium microprocessors capable of running the image analysis software.
- One software package suitable for the present image analysis is Global Lab Image Version 3.1, available from Data Translation.
- Equation 1 gives a formula which has been used previously to obtain the average cell diameter in terms of the radius (r) of the first ring of light scattered at wavelength (2) when focussed with a camera lens of focal length (f).
- Equation 2 gives a formula relating the cell concentration (c) in terms of the
- equation 1 the result from equation 1 can be used in equation 2, eliminating the ambiguity generally associated with estimates of cell concentration due to the size of the particles.
- the time variation in intensity of the scattered light may be used to determine the rate of aggregation and sedimentation.
- Two effects govern this determination; first, there is a steady increase in light transmission (reduction in turbidity) as a greater proportion of the RBC that are hidden by their neighbors, and second, there is an increase in the fluctuation of intensity due to coherent interference between the scattering from individual RBC. These contributions can be used to estimate the erythrocyte sedimentation rate (ESR).
- ESR erythrocyte sedimentation rate
- the sample apparatus as illustrated previously may be used. However, in this case the fluid used to obtain the low dilution of whole blood must contain an agent which hemolyzes the RBC and destroys their membrane. Phthalic acid at 21.5mmol may be
- the dominant scattering cell population is the WBC.
- the first angular interval is preferably selected to span the angular region within which the first maxima of the light scattering patterns of the cell populations having volumes in the physiological range are expected to fall.
- the second angular interval is selected which begins above the upper limit of the first angular interval and below the second maximum of the scattering pattern of the cell population.
- fractional cell concentrations of e.g. PLT/RBC, GRAN/ WBC, GRAN/AGRAN, MONO/POLY or POLY/WBC
- a physical mask is employed to isolate the region of scattering that differentiates the population of interest, i.e. for platelets which are smaller than RBC one might isolate the high angles of scatter in comparison with the value for the total scattering.
- a more specific estimate of the PLT/RBC fraction can be made by using a mask which is the inverse of the RBC scattering pattern.
- the light that gets through the mask corresponds to some other cell type such as PLT. Comparing intensity with and without such a mask estimates the concentration of PLT.
- a virtual (hypothetical with values) or algorithmic mask may be entered into the image analysis stage to achieve the same effect as interposing the physical mask before the camera.
- the digital data corresponding to the ensemble scattering of the cell population will be filtered to enhance the contribution of a particular constituent, e.g. RBC, WBC, AGRAN or POLY.
- values for turbidity or light scattering intensity are determined as a function of time. Altematively, the fluctuations in intensity can be measured as a function of time.
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP9522913A JP2000502446A (en) | 1995-12-18 | 1996-12-12 | Blood diagnostic apparatus and determination method |
EP96943726A EP0888531A1 (en) | 1995-12-18 | 1996-12-12 | Multi-parameter hematology apparatus and method |
AU12884/97A AU1288497A (en) | 1995-12-18 | 1996-12-12 | Multi-parameter hematology apparatus and method |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US881895P | 1995-12-18 | 1995-12-18 | |
US60/008,818 | 1995-12-18 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1997022870A1 true WO1997022870A1 (en) | 1997-06-26 |
Family
ID=21733847
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1996/019823 WO1997022870A1 (en) | 1995-12-18 | 1996-12-12 | Multi-parameter hematology apparatus and method |
Country Status (5)
Country | Link |
---|---|
US (1) | US5851835A (en) |
EP (1) | EP0888531A1 (en) |
JP (1) | JP2000502446A (en) |
AU (1) | AU1288497A (en) |
WO (1) | WO1997022870A1 (en) |
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1996
- 1996-12-12 WO PCT/US1996/019823 patent/WO1997022870A1/en not_active Application Discontinuation
- 1996-12-12 AU AU12884/97A patent/AU1288497A/en not_active Abandoned
- 1996-12-12 US US08/764,322 patent/US5851835A/en not_active Expired - Fee Related
- 1996-12-12 JP JP9522913A patent/JP2000502446A/en active Pending
- 1996-12-12 EP EP96943726A patent/EP0888531A1/en not_active Withdrawn
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EP0507746A2 (en) * | 1991-04-03 | 1992-10-07 | Istituto Nazionale Di Ottica | Method and device for measuring the particle size distribution of a flowing solid particulate substance |
EP0575712A2 (en) * | 1992-03-31 | 1993-12-29 | University Of Manitoba | Spectrophotometric blood analysis |
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---|---|---|---|---|
EP1106991A2 (en) * | 1999-12-01 | 2001-06-13 | Fresenius HemoCare GmbH | Method and apparatus for determining particle concentration |
EP1106991A3 (en) * | 1999-12-01 | 2001-12-19 | Fresenius HemoCare GmbH | Method and apparatus for determining particle concentration |
US6627911B2 (en) | 1999-12-01 | 2003-09-30 | Fresenius Hemocare Gmbh | Method for determining a particle concentration and device for implementing the method |
WO2002088676A1 (en) * | 2001-04-26 | 2002-11-07 | Evotec Oai Ag | Method and device for detecting the state of objects |
Also Published As
Publication number | Publication date |
---|---|
JP2000502446A (en) | 2000-02-29 |
US5851835A (en) | 1998-12-22 |
EP0888531A1 (en) | 1999-01-07 |
AU1288497A (en) | 1997-07-14 |
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