US3916205A - Differential counting of leukocytes and other cells - Google Patents
Differential counting of leukocytes and other cells Download PDFInfo
- Publication number
- US3916205A US3916205A US365460A US36546073A US3916205A US 3916205 A US3916205 A US 3916205A US 365460 A US365460 A US 365460A US 36546073 A US36546073 A US 36546073A US 3916205 A US3916205 A US 3916205A
- Authority
- US
- United States
- Prior art keywords
- dyes
- leukocytes
- cell
- exposures
- fluorescent
- Prior art date
- Legal status (The legal status 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 status listed.)
- Expired - Lifetime
Links
- 210000000265 leukocyte Anatomy 0.000 title claims abstract description 123
- 210000004027 cell Anatomy 0.000 title claims abstract description 71
- 239000000975 dye Substances 0.000 claims abstract description 112
- 239000007850 fluorescent dye Substances 0.000 claims abstract description 27
- 239000000203 mixture Substances 0.000 claims abstract description 26
- 210000001995 reticulocyte Anatomy 0.000 claims abstract description 20
- 210000003743 erythrocyte Anatomy 0.000 claims abstract description 19
- 238000010186 staining Methods 0.000 claims abstract description 19
- 210000004369 blood Anatomy 0.000 claims abstract description 17
- 239000008280 blood Substances 0.000 claims abstract description 17
- 238000010521 absorption reaction Methods 0.000 claims description 37
- 238000000034 method Methods 0.000 claims description 31
- 210000003979 eosinophil Anatomy 0.000 claims description 23
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 claims description 18
- 229960005542 ethidium bromide Drugs 0.000 claims description 18
- 239000008187 granular material Substances 0.000 claims description 18
- 210000000440 neutrophil Anatomy 0.000 claims description 18
- 210000000805 cytoplasm Anatomy 0.000 claims description 16
- 210000001616 monocyte Anatomy 0.000 claims description 16
- 210000004698 lymphocyte Anatomy 0.000 claims description 14
- 230000004044 response Effects 0.000 claims description 14
- 230000005855 radiation Effects 0.000 claims description 13
- 125000004306 triazinyl group Chemical group 0.000 claims description 13
- 238000005286 illumination Methods 0.000 claims description 12
- HYLDLLCHFLSKAG-UHFFFAOYSA-M lissamine flavine FF Chemical group [Na+].C1=CC(C)=CC=C1N(C1=O)C(=O)C2=C3C1=CC=CC3=C(N)C(S([O-])(=O)=O)=C2 HYLDLLCHFLSKAG-UHFFFAOYSA-M 0.000 claims description 10
- 210000000601 blood cell Anatomy 0.000 claims description 9
- 230000003287 optical effect Effects 0.000 claims description 8
- 230000005540 biological transmission Effects 0.000 claims description 6
- 102000039446 nucleic acids Human genes 0.000 claims description 4
- 108020004707 nucleic acids Proteins 0.000 claims description 4
- 150000007523 nucleic acids Chemical class 0.000 claims description 4
- 102000004169 proteins and genes Human genes 0.000 claims description 4
- 108090000623 proteins and genes Proteins 0.000 claims description 4
- TXVWTOBHDDIASC-UHFFFAOYSA-N 1,2-diphenylethene-1,2-diamine Chemical compound C=1C=CC=CC=1C(N)=C(N)C1=CC=CC=C1 TXVWTOBHDDIASC-UHFFFAOYSA-N 0.000 claims description 3
- XJHABGPPCLHLLV-UHFFFAOYSA-N benzo[de]isoquinoline-1,3-dione Chemical class C1=CC(C(=O)NC2=O)=C3C2=CC=CC3=C1 XJHABGPPCLHLLV-UHFFFAOYSA-N 0.000 claims description 3
- 241000269627 Amphiuma means Species 0.000 claims description 2
- 230000002378 acidificating effect Effects 0.000 claims description 2
- FDXOYIGOUGEQBC-UHFFFAOYSA-N 8-(4-methylanilino)naphthalene-1-sulfonic acid Chemical compound C1=CC(C)=CC=C1NC1=CC=CC2=CC=CC(S(O)(=O)=O)=C12 FDXOYIGOUGEQBC-UHFFFAOYSA-N 0.000 claims 1
- 230000003595 spectral effect Effects 0.000 abstract description 8
- 230000001678 irradiating effect Effects 0.000 abstract description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 12
- 230000005284 excitation Effects 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 9
- 239000000725 suspension Substances 0.000 description 8
- 125000003545 alkoxy group Chemical group 0.000 description 7
- 125000000217 alkyl group Chemical group 0.000 description 7
- 229910052736 halogen Chemical group 0.000 description 7
- 150000002367 halogens Chemical group 0.000 description 7
- 125000000954 2-hydroxyethyl group Chemical group [H]C([*])([H])C([H])([H])O[H] 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 239000000872 buffer Substances 0.000 description 6
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 6
- 229910052753 mercury Inorganic materials 0.000 description 6
- PSZYNBSKGUBXEH-UHFFFAOYSA-N naphthalene-1-sulfonic acid Chemical compound C1=CC=C2C(S(=O)(=O)O)=CC=CC2=C1 PSZYNBSKGUBXEH-UHFFFAOYSA-N 0.000 description 5
- BIEFDNUEROKZRA-UHFFFAOYSA-N 2-(2-phenylethenyl)aniline Chemical compound NC1=CC=CC=C1C=CC1=CC=CC=C1 BIEFDNUEROKZRA-UHFFFAOYSA-N 0.000 description 4
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- -1 4-(3- sulfoanilino)-6-[bis (2-hydroxyethyl)-amino]- 1,3,5triazin-2-yl amino stilbene 2,2-disulfonic acid tetrasodium salt Chemical compound 0.000 description 3
- PJANXHGTPQOBST-VAWYXSNFSA-N Stilbene Natural products C=1C=CC=CC=1/C=C/C1=CC=CC=C1 PJANXHGTPQOBST-VAWYXSNFSA-N 0.000 description 3
- 235000021286 stilbenes Nutrition 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- DPKHZNPWBDQZCN-UHFFFAOYSA-N acridine orange free base Chemical compound C1=CC(N(C)C)=CC2=NC3=CC(N(C)C)=CC=C3C=C21 DPKHZNPWBDQZCN-UHFFFAOYSA-N 0.000 description 2
- 125000002490 anilino group Chemical group [H]N(*)C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 2
- 229910021538 borax Inorganic materials 0.000 description 2
- 125000002091 cationic group Chemical group 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000001747 exhibiting effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 235000010339 sodium tetraborate Nutrition 0.000 description 2
- PJANXHGTPQOBST-UHFFFAOYSA-N stilbene Chemical compound C=1C=CC=CC=1C=CC1=CC=CC=C1 PJANXHGTPQOBST-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- KBVDUUXRXJTAJC-UHFFFAOYSA-N 2,5-dibromothiophene Chemical compound BrC1=CC=C(Br)S1 KBVDUUXRXJTAJC-UHFFFAOYSA-N 0.000 description 1
- CBTLENLQSONRNU-UHFFFAOYSA-N 4-(2-phenylethenyl)cyclohexa-2,4-diene-1,1-diamine Chemical class C1=CC(N)(N)CC=C1C=CC1=CC=CC=C1 CBTLENLQSONRNU-UHFFFAOYSA-N 0.000 description 1
- ZFXPBTZXYNIAJW-UHFFFAOYSA-N 4-[2-(2-phenylethenyl)phenyl]triazine Chemical class C=1C=CC=CC=1C=CC1=CC=CC=C1C1=CC=NN=N1 ZFXPBTZXYNIAJW-UHFFFAOYSA-N 0.000 description 1
- 241000237519 Bivalvia Species 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 210000003651 basophil Anatomy 0.000 description 1
- DZBUGLKDJFMEHC-UHFFFAOYSA-N benzoquinolinylidene Natural products C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000020639 clam Nutrition 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- BFMYDTVEBKDAKJ-UHFFFAOYSA-L disodium;(2',7'-dibromo-3',6'-dioxido-3-oxospiro[2-benzofuran-1,9'-xanthene]-4'-yl)mercury;hydrate Chemical compound O.[Na+].[Na+].O1C(=O)C2=CC=CC=C2C21C1=CC(Br)=C([O-])C([Hg])=C1OC1=C2C=C(Br)C([O-])=C1 BFMYDTVEBKDAKJ-UHFFFAOYSA-L 0.000 description 1
- 239000010408 film Substances 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000006193 liquid solution Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- GKZKFYQHCPQNNG-UHFFFAOYSA-N propane-1,1-diol;hydrochloride Chemical compound Cl.CCC(O)O GKZKFYQHCPQNNG-UHFFFAOYSA-N 0.000 description 1
- ORUDTFXLZCCWNY-UHFFFAOYSA-N pyrene-1,2,3-trisulfonic acid Chemical class C1=CC=C2C=CC3=C(S(O)(=O)=O)C(S(=O)(=O)O)=C(S(O)(=O)=O)C4=CC=C1C2=C43 ORUDTFXLZCCWNY-UHFFFAOYSA-N 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000004328 sodium tetraborate Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- BSVBQGMMJUBVOD-UHFFFAOYSA-N trisodium borate Chemical compound [Na+].[Na+].[Na+].[O-]B([O-])[O-] BSVBQGMMJUBVOD-UHFFFAOYSA-N 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
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
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1468—Electro-optical investigation, e.g. flow cytometers with spatial resolution of the texture or inner structure of the particle
- G01N15/147—Electro-optical investigation, e.g. flow cytometers with spatial resolution of the texture or inner structure of the particle 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
- G01N15/1468—Electro-optical investigation, e.g. flow cytometers with spatial resolution of the texture or inner structure of the particle
-
- G01N2015/016—
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
- G01N2021/6441—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks with two or more labels
Definitions
- the present invention relates to cytology and, more particularly is directed towards a composition for staining specific components of cells particularly blood cells,and a method and apparatus for differentially counting and classifying leukocyte types.
- cytotechnician microscopically views ablood smear prepared on an ordinary microscope slide that has been stained with one of the Romanowsky stains, such as'the Wright or Giemsa stains.
- the cytotechnician sequentially examines I leukocytes and classifies each accordingly to its type. Not only is this method time consuming, but it suffers also from the disadvantage of limited reliability relative to counting and classifying of the less abundant cells such as monocytes, eosinophils and basophils.
- the leukocytes are made to flow through three or four different channels, each channel provided with means for staining the leukocytes flowing therein with a particular dye. Due to the fact that several channels and difl'erent chemical treatments are required for each cell type, such a system sufiers from thedisadvantages that it is complex in design and costly to manufacture.
- a selected region of a blood smear stained in the usual manner with a Romanowsky stain, is scanned mechanically under a microscope provided with an electronic image tube.
- an image analyzing computer connected to the image tube classifies the leukocyte according to its cell profile and cytoplasm color. Since this system requires'the services of a technician to select a region of the smear suitable for automatic scanning, such systems have suffered from the disadvantages of being time consumming and costly.
- the present invention provides a dye com position for distinguishing eosinophils, monocytes, lymstained leukocytes are irradiated with optical radiation having the characteristic absorption wavelengths of each dye, the relative intensities of the light emitted and/or by each leukocyte defines 'a
- compatible d'yes is used here to mean that no dye in the mixture prevents any other dye in the mixture from selectively staining characteristic structures of the cell types of interest when irradiated in the characteristic absorption region of said other dye, such selective staining being essentially not different from the action of said other dye when used by itself.
- fluorochrome stain is used to designate a stain which under suitable illumination produces better differentiated optical parameters about the cell type of interest by its fluorescence than by its absorption properties, for instance, when the light transmitted by the dye in its main absorption band is not much different in intensity from that transmitted by its surroundings, but the fluorescence intensity of the same dye is several times or more greater than that of its surroundings.
- differential staining is used to mean that the dye being used will stain a given component of the cell type of interest for instance the protein of said cell type to a much greater extent that other components, for instance nucleic acids, of the same cell type.
- One fluorescent dye composition particularly adapted for slide scanning and flow tube methods for diflerentially counting and classifying leukocytes, comprises sulfonated triazinyl derivatives of a diarnino stilbene, having each amino group in each of the phenyl rings of the stilbene molecule, preferably a sulfonated 4,4 diamino stilbene, either of the dyes 8-p-toluidino-lnaphthalene sulfonic acid or brilliant sulfaflavine, and ethidium bromide.
- a drop of blood is spread on a microscope slide and, after fixation, is stained by immersion in the dye composition.
- a small amount of a blood sample is stained in a liquid solution of the dye composition and the cells are forced to flow in single file through a capillary tube.
- the stained leukocytes are irradiated under ultraviolet and violet light and their fluorescence is detected through a blue-green filter.
- the leukocytes are presented with a fluorescence in decreasing order to intensity from eosiniphils to immature neutrophils to mature neutro phills to monocytes to lymphocytes.
- the stained leukocytes are then irradiated under a greenlight and the nuclei of the different leukocyte types are detected through a red filter.
- Leukocytes exhibit a red fluorescence of comparable intensity, reticulocytes fluoresce with an appreciably smaller intensity and normal erythrocytes have a negligible fluorescence.
- the leukocytes are counted and classified, for example, by measuring the ratio of the blue-green component of cell fluorescence to the red nuclear fluorescence.
- excitation with ultraviolet light produces a strong orange-red component, most pronounced in neutrophils and eosinophils, the intensity of which, with respect to the blue-green component, provides an- 7 other parameter for distinguishing particular leukocyte types.
- the transmitted violet light is measured to distinguish erythrocytes from leukocytes. In contrast to the negligible absorption of violet light by leukocytes, erythrocytes and reticulocytes exhibit an appreciable absorption of violet light.
- the invention accordingly comprises the composition, method steps and apparatus possessing the construction, combination of elements, and arrangement of parts that are exemplified in the following detailed disclosure, the scope of which will be indicated in the appended claims.
- FIG. 1 is a block and schematic diagram of a slide scanning system for differential counting of leukocytes
- FIG. 2 is a block and schematic diagram of a flow tube system for differential counting of leukocytes.
- FIG. 3 is a graphic representation illustrating the fluorescent characteristics of selected leukocyte types on a methanol fixed smear stained with one of the mixtures herein described.
- the process embodying the present invention involves the staining of a blood medium with a dye composition comprising a plurality of dyes, at least one of the dyes imparting a characteristic fluorescence to all leukocytes.
- the dye composition is such that, when the stained blood medium is illuminated with light having the characteristic absorption wavelengths of each dye, the relative intensities of the light emitted and/or transmitted by each cell type at the optical wavelength region of emission and absorption characteristic of each dye uniquely depends, in at least one characteristic wavelength region, on the type of leukocyte.
- the light emitted by the irradiated leukocytes in each of their characteristic wavelength regions is measured and the leukocyte types are classified according tothe relative intensities of the emitted light in the characteristic wavelength region of each dye.
- the process embodying the present invention involves the staining of leukocytes randomly distributed in a medium with a dye composition comprising a mixture of (l) a sulfonated triazinyl stilbene derivative, (2) a naphthalene sulfonic acid; and (3) a cationic dye.
- the stained leukocytes are irradiated with ultraviolet and violet light and detected through a green or blue-green filter.
- the green component of leukocyte fluorescence is presented with a characteristic fluorescent intensity against a dark background.
- the stained leukocytes are also irradiated with green light and the red fluorescence of only the nuclei of the cells are detected through a red or orange-red filter. As shown in FIG.
- the detected leukocytes are characterized by a fluorescence in decreasing order of intensity from eosinophils to neutrophils to monocytes to lymphocytes, each leukocyte type uniquely defined by the ratio of the blue-green component of cell fluorescence to the red nuclear fluorescence with respect to the red fluorescence from the nucleus.
- the dyes for protein such as the cytoplasm of white blood cells and granules of granulocytes, preferably are sulfonated triazinyl derivatives of a diamino stilbene, for example 4,4-bis 4-(3 sulfoanilino)-6-[bis (2- hydroxy-ethyl)-amino]-l ,3,5triazin-2-yl amino stilbene 2,2disulfonic acid tetrasodium salt and their alkyl, alkoxy or halogen substituted derivatives.
- the optimum excitation wavelength band is in the range of 320 to 390 nanometers and the optimum fluorescent wavelength band is 440 to 550 nanometers.
- These dyes impart a strong blue fluorescence to the protein of leukocytes and the neutrophil granules and a weaker blue fluorescence to eosinophils, monocytes and lymphocytes.
- the dyes for eosinophil granules preferably are the anilino or toluidino naphthalene sulfonic acids and their alkyl, alkoxy or halogen substituted derivatives;
- 4,4 diamino stilbene 2,2 disulfonic acid N, N, N, N' tetraacetric acid and its alkyl, alkoxy or halogen substituted derivatives
- sulfonated fluorescent derivatives of 1,8 naphthalimide such as'brilliant s'ulfaflavine and its alkyl, alkoxy or halogen substituted derivatives
- the anilino and toluidino naphthalene sulfonic acids have an optimum excitation wavelength band in the range of 320 to 410 nanometers and an optimum fluorescent wavelength band in the range of 440 to 550 nanometers.
- Brilliant sulfoflavine has an optimum excitation wavelength band in the range of 360 to 450 nanometers and an optimum fluorescent wavelength band in the range of 480 to 550 nanome-- ters. These dyes strongly stain the'eosinophil granules with a green-blue or green fluorescence.
- the dyes which impart a strong red fluorescence to the nucleic acids of all leukocytes, mainly in the nuclei, preferably are cationic dyes, for example the phenanthridinium dyes such as ethidium bromide and its alkyl, alkoxy or halogen derivatives; and in dry smears, acridine orange; and rhoduline orange.
- Ethidium bromide has an optimum excitation wavelength range of 480 to 550 nanometers and an optimum fluorescent wavelength range of 580 to 650 nanometers.
- a slide scanning system 10 for differentially counting leukocytes.
- an alcohol-fixed blood smear 11 on a slide 12 is immersed in a preferred dye solution comprising 1 X 10 molar ethidium bromide; l X 10' to l X 10 molar brilliant sulfaflavine; andl X 10 molar 4,4 bis 4-(3-sulfoanilino)-6[bis (2-hydroxyethyl)amino]-l,3,5 triazin-Z-yl amino stilbene 2,2 disulfonic acid tetrasodium salt in a solvent including a buffer of 0.01 to 0.1 molar 2-amino-2(hydroxymethyl)-l,3 propanediol-HCI, commonly known as tris- HCl, at a pH concentration range of 8.0 to 10.0, for best results, the pH concentration range is 8.5 to 9.5.
- the buffer is other than tris-I-ICl, for example sodium borate or sodium bicarbonate.
- slide 12 is rinsed in an aqueous solution, for example distilled water, for approximately 1 minute and then dried.
- slide 12 is irradiated with illumination from a source 14, for example a mercury arc lamp, which is focused thereon via a collimating lens 16, an excitation filter wheel 18, a beam splitter 20 and a condensing lens 22.
- Filter wheel 18 includes a filter 32 which transmits ultraviolet and violet light, for example 320 to 440 nanometers and a filter 34 which transmits green light, for example the 546 nanometer mercury band.
- beam splitter 20 is a dichroic mirror.
- beam splitter 20 is other than a dichroic mirror, for example a reflector such as a straight surface rnirror.
- the field of view containing the fluorochromed leukocytes is imaged on a photo-electronic device 24, for example the photosensitive surface of an image scanning tube, via a collimating lens 26 and an emission filter wheel 28.
- Filter wheel 28 includes a blue-green filter 36 and a red filter 38. As hereinafter described, filter wheels 18 and 28 are indexed by means of a controller 30.
- controller 30 indexes filter wheels 18 and 28 insuch a manner that filters 32 and 36 are positioned -in the transmitted light path. That is, slide 12 is irradi- ,ated with ultraviolet and violet light and blue-green fluorescent images are presented at the photosensitive .eosiniphils and neutrophils.
- the photosensitive surface of image tube 24 is scanned in a specified pattern determined by a control 40 which is programmed by a computer 42.
- the intensity of the irradiated leukocytes are measured by computer 42 and data signals for each measurement are stored in a memory 44 at X,Y address location corresponding to the X,Y positions on the photosensitive surface of image scanning tube 24.
- Controller 30 then indexes filter wheels 18 and 28 in such a manner that filters 34 and 38 are positioned in the transmitted light path. That is, slide 12 is irradiated with green light and red fluorescent images are presented at the photosensitive surface of image scanning tube 24 through red filter 38. In this case, only the red fluorescence of the nuclei of the leukocytes appear against a dark background at the photosensitive surface of image tube 24; In the manner hereinbefore described, the fluorescent intensities of the nuclei at each of the X,Y positions on the photosensitive surface of image scanning tube 24 are measured by computer 42.
- the measurement data stored'in memory 44 i.e., the
- a green component of cell fluorescence is addressed into computer42 for determining leukocyte types by generating differential counting data signals representing the ratio of the green component of cell fluorescence to red nuclear fluorescence with respect to the red fluorescence from the nucleus as shown in FIG. 3.
- Cell profile data signals i.e. ameasurement of the time period during which a signal from a cell is received, distinguish lymphocytes from monocytes.
- the differential counting data signals generated by computer 42 are applied to a display 46, for example a digital display of the relative abundance of the different cell types, for visual presentation. It is to be understood that, in alternative embodiments display 46 is other than a digital display, for example a cathode-ray tube, a chart recorder, or a magnetic tape recorder. 7
- systems other than that shown in FIG. 1 can be used for differentially counting and classifying leukocytes types on a stained blood smear.
- a manual system wherein the stained slide is irradiated in the manner hereinbefore described and observed through a microscope.
- the irradiated leukocytes are detected by means other than an image scanning tube, for example one or a plurality of photo-detectors and a flying spot scanner.
- a flow tube system 50 for differential counting of leukocyte types By way of example, a blood sample is diluted in a saline solution and the cells are fixed with formaldehyde. It is to be noted that the cells can be fixed with formaldehyde prior to or after dilution.
- the white cells are stained in a solution comprising 1 X molar 4,4 bis I 4-(3 sulfoanilino)-6-[bis (2-hydroxy ethyl) amino]- 1,3,5 triazin -2- yl ⁇ amino stilbene; 1 X 10 to l X 10 molar brilliant sulfaflavine; and l X 10 molar ethidium bromide.
- the resulting suspension is diluted and buffered with either 0.1 molar tris-HCl or 0.05 molar borax at a pH in the range of 9.0 to 9.2.
- the blood sample has a dilution range. of fifty to one hundred fold in the suspension, the final concentration of which is approximately 1 X 10 molar or less in each of the fluorescent dyes.
- the cells. are made to flow single file through a narrow tube 52, for example a capillary tube, wherein each of the leukocytes is irradiated with illumination from a source 54, for example a mercury arc lamp.
- the illumination generated by mercury arc lamp 54 is directed through a collimating lens 56 to a reflecting filter 58, for example a dichroic filter.
- dichroic filter 58 transmits green light in the 546 nanometer mercury band.
- the wavelength band of light reflected by dichroic filter 58 is directed to a reflecting filter 66, for example a dichroic filter, and focused on capillary tube 52 at via a condensing lens 72.
- dichroic filter 66 reflects ultraviolet and violet light in the 320 to 410 nanometer mercury band. It is preferred that the distance between the locations denoted by reference characters 62 and 70 is typically in the range of 50 to 200 microns.
- a photo-electric device 74 for example a photo-multiplier senses the blue-green emission light from capillarytube 52 via a collecting lens 76, a dichroic mirror 77 and a blue-green emission filter 78.
- a photo-electric device 80 for example a photo-multiplier, senses the orange-red emission light from capillary tube 52 via collecting lens 76, dichroic mirror 77, a dichroic mirror 82 and an orange-red emission filter 84.
- a photo-electric device 81 for example a photodiode, senses the ultraviolet and violet light at 70 via an emission filter 82 having a pass band in the approximaterange of 410 to 430 nanometers.
- Photo-multiplier 74 detects the blue-green component of cell fluorescence
- photo-multiplier 80 detects the red fluorescence
- photo-multiplier 81 detects the violet light.
- Erythrocytes and reticulocytes are characterized by appreciable absorption of violet light and leukocytes are characterized by negligible absorption of violet light. Accordingly, photo-multiplier 81 detects the violet light absorption for distinguishing red cells from white cells.
- Data signals generated by photo-multipliers 74 and 80 are applied to a precessor 86, for example a small dedicated computer, wherein the detected leukocyte fluorescences are differentially counted and classified as the ratio of the green component of cell fluorescence to the red nuclear fluorescence with respect to the red fluorescence from the nucleus for each leukocyte type.
- display 88 is other than a digital display for example, a cathode-ray tube, a chart recorder or a magnetic tape recorder.
- the light emitted from source 54 is directed to tube 52 by means other than a pair of beam splitters, for example, two sources each emitting light which is directed towards tube 52 or a source characterized by a scanning light beam.
- EXAMPLE I SLIDE A drop of whole blood is placed on a microscope slide and spread into a thin film with the aid of another microscope slide. After the film has dried, the blood smear is fixed by immersing the slide in methyl alcohol for five minutes. Thereafter, the slide is immersed in a liquid mixture comprising:
- the slide After ten minutes in the liquid mixture, the slide is rinsed in distilled water for one minute and then dried.
- the stained suspended cells exhibit the following optical properties under ultraviolet illumination of approximately 365 nanometers:
- the cytoplasm and granules of the neutrophils exhibit a visible fluorescence with spectral peaks in the blue and orange-red regions.
- the blue component results from the direct excitation of the triazinyl dye and the red component, most pronounced in the granules, is due to energy transfer from the triazinyl dye to ethidium bromide, the latter being present in the granules, and elsewhere in the cytoplasm, at concentrations which are too small to be efficienctly excited by direct absorption of the ultraviolet light.
- the cytoplasm and granules of the eosinophils exhibit a stronger visible fluorescence than the neutrophils with spectral peaks in the blue-green and orange-red regions.
- the blue-green component results mainly from the excitation of the brilliant sulfaflavine dye and the triazinyl dye, and the redorange component is due to energy transfer from the triazinyl dye and the brilliant sulfaflavine dye to ethidium bromide; the latter being present in the granules, and elsewhere in the cytoplasm at concentrations too small to be efficiently excited by direct absorption of the ultraviolet light.
- the cytoplasm and nucleus of lymphocytes and monocytes exhibit a weaker fluorescence than that of either the neutrophils or eosinophils.
- This fluorescence is characterized by a spectral distribution having a peak in the blue region and a smaller peak or shoulder in the red region, the fluorescence of the monocytes being smaller than the fluorescence of the lymphocytes.
- the eosinophils Under violet illumination of approximately 400 to 440 nanometers, the eosinophils exhibit a strong green fluorescence which is several times stronger than that of the other leukocytes.
- the nuclei, and to a lesser extent the cytoplasm, of all the leukocytes exhibit a red fluorescence; eosinophils having the brightest fluorescent intensity, neutrophils and monocytes having comparable fluorescent intensities of a lesser brilliance, and
- lymphocytes generally having the lowest fluorescent intensity.
- red cells exhibit a negligible fluorescence under any of the above conditions.
- the stained suspended cells exhibit the following optical properties under ultraviolet illumination of approximately 365 nanometers:
- cytoplasm and granules of the neutrophils ex-' hibit a visible fluorescence with spectral peaks in the blue and orange-red regions.
- the blue component results from the direct excitation of the triazinyl dye and the red component, most pronounced in the granules, is due to energy transfer from the triazinyl dye to ethidium bromide, the latter being present in the granules, and elsewhere in the cytoplasm, at concentrations which are too small to be efficiently excited by direct absorption of the ultraviolet light.
- the cytoplasm and granules of the eosinophils exhibit a stronger visible fluorescence than the neutrophils with spectral peaks in the blue-green and orange-red regions.
- the blue-green component results mainly from the excitation of 8-p-toluidino-lnaphthalene sulfonic acid and the triazinyl dye, and the red-orange component is due to energy transfer from the triazinyl dye and the 8-ptoluidino-lnaphthalene sulfonic acid to ethidium bromide; the latter being present in the granules, and elsewhere in the cytoplasm at concetrations too small to be efficiently excited by direct absorption of the ultraviolet light.
- the cytoplasm and nuclei of lymphocytes and monocytes exhibit a weaker fluorescence than that of either the neutrophils or eosinophils.
- This fluorescence is characterized by a spectral distribution having a peak in the blue region and a smaller peak or shoulder in the red region, the fluorescence of the monocytes being smaller than the fluorescence of the lymphocytes.
- the eosinophils Under violet illumination of approximately 400 to 440 nanometers, the eosinophils exhibit a strong green fluorescence which is several times stronger than that of the other leukocytes.
- the nuclei, and to a lesser extent the cytoplasm, of all the leukocytes exhibit a red fluorescence; eosinophils having the brightest fluorescent intensity, neutrophils and monocytes having comparable fluorescent intensities of a lesser brilliance, and lymphocytes generally having the lowest fluorescent intensity.
- red cells exhibit a negligible fluorescenceunder any of the above conditions.
- the present invention provides a single dye composition comprising three fluorescent dyes for uniquely distinguishing leukocytes, erythrocytes and reticulocytes randomly distributed in a medium and a method and apparatus for absolute counting of leukocytes and reticulocytes and differential counting of leukocytes which have been stained with such a dye composition.
- the stained leukocytes are irradiated by light having the characteristic absorption wavelengths of each dye, the relative intensities of leukocyte fluorescence at the optical wavelength region of emission of each dye depends, in at least one characteristic wavelength region, on the type of leukocyte.
- the light emitted by the irradiated leukocytes in each of their characteristic wavel'ength regions is measured and the leukocyte types are classified according to the relative intensities of the emitted light in the characteristic wavelength region of each dye.
- Transmitted violet light is measured to distinguish erythrocytes, reticulocytes and leukocytes, erythrocytes and reticulocytes exhibiting appreciable absorption of violet light and leukocytes exhibiting negligible absorption of violet light.
- At least one of which is a fluoresecent dye illuminating the dyed cell with a sequence of expo sures to radiation, each of said exposures being at a limited wavelength band corresponding to the wavelength absorption band of a respective one of said dyes;
- a system for identifying leukocytes, erythrocytes and reticulocytes randomly distributed in a medium comprising:
- source means for sequentially illuminating the stained leukocytes, erythrocytes and reticulocytes with optical radiation in the characteristic absorption wavelength band of each said dye and c. detector means optically coupled to said irradiated leukocytes, erythrocytes and reticulocytes for identifying each said type cell according to its fluorescent intensity.
- said detector means includes photodetector means for measuring light scattered by said leukocytes for differentiating said leukocytes according to their size.
- detector means optically coupled to said slide means for detecting, during each of said exposures, the characteristic wavelength band emitted by one of said dyes in said leukocytes in response to a corresponding one of said exposuresbut not the characteristic wavelength band emitted by any of the other dyes responsively to said one of said exposures;
- a system for differentially classifying and counting leukocyte types randomly distributed in a blood sample comprising:
- a dye composition comprising a plurality of fluorescent dyes, at least one of said fluorescent dyes imparting a first characteristic fluorescence to the nuclei of all said leukocytes, said other fluorescent dyes imparting a second characteristic fluorescence to the cytoplasm of eosinophils, neutrophils, monocytes and lymphocytes;
- said means for illuminating includes means for providing at least two of said exposures at different wavelength bands corresponding to the absorption bands of respective ones of said dyes; and i said means for observing includes means for observing fluorescent emission in the wavelength band of emission from said fluorescent dye dueonly to exposure of said cell to radiation in the wavelength absorption band of said fluorescent dye, and means for observing fluorescent emission in the wavelength band of emission from said fluorescent dye due only to exposure of said cell to radiation in the wavelength absorption band of another of said dyes.
Abstract
Description
Claims (18)
Priority Applications (10)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US365460A US3916205A (en) | 1973-05-31 | 1973-05-31 | Differential counting of leukocytes and other cells |
US05/438,162 US4146604A (en) | 1973-05-31 | 1974-01-30 | Differential counting of leukocytes and other cells |
FR7411363A FR2231968B1 (en) | 1973-05-31 | 1974-03-29 | |
GB1723474A GB1475317A (en) | 1973-05-31 | 1974-04-19 | Method and composition for staining biological material |
GB4106676A GB1475318A (en) | 1973-05-31 | 1974-04-19 | Method for differentially counting leukocytes and other cells |
DE2421501A DE2421501A1 (en) | 1973-05-31 | 1974-05-03 | METHOD AND DEVICE FOR ANALYSIS OF LEUKOCYTE AND SIMILAR CELLS |
AU68612/74A AU6861274A (en) | 1973-05-31 | 1974-05-06 | Biological stain for counting leukocytes |
NL7406331A NL7406331A (en) | 1973-05-31 | 1974-05-10 | |
CH660074A CH582355A5 (en) | 1973-05-31 | 1974-05-14 | |
JP49054272A JPS5020820A (en) | 1973-05-31 | 1974-05-15 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US365460A US3916205A (en) | 1973-05-31 | 1973-05-31 | Differential counting of leukocytes and other cells |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US05/438,162 Continuation-In-Part US4146604A (en) | 1973-05-31 | 1974-01-30 | Differential counting of leukocytes and other cells |
Publications (1)
Publication Number | Publication Date |
---|---|
US3916205A true US3916205A (en) | 1975-10-28 |
Family
ID=23439004
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US365460A Expired - Lifetime US3916205A (en) | 1973-05-31 | 1973-05-31 | Differential counting of leukocytes and other cells |
Country Status (1)
Country | Link |
---|---|
US (1) | US3916205A (en) |
Cited By (86)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3992631A (en) * | 1975-02-27 | 1976-11-16 | International Diagnostic Technology, Inc. | Fluorometric system, method and test article |
US4000417A (en) * | 1975-08-25 | 1976-12-28 | Honeywell Inc. | Scanning microscope system with automatic cell find and autofocus |
US4015595A (en) * | 1975-09-15 | 1977-04-05 | Benjamin Jr J Malvern | Photoplethysmographs |
US4025310A (en) * | 1976-05-28 | 1977-05-24 | International Diagnostic Technology, Inc. | Method for reading a wet fluorescent surface |
US4071891A (en) * | 1976-04-30 | 1978-01-31 | Barrows George H | Electronic calculator - register for hematology differentials |
US4097845A (en) * | 1976-11-01 | 1978-06-27 | Rush-Presbyterian-St. Luke's Medical Center | Method of and an apparatus for automatic classification of red blood cells |
US4117338A (en) * | 1977-05-24 | 1978-09-26 | Corning Glass Works | Automatic recording fluorometer/densitometer |
US4125828A (en) * | 1972-08-04 | 1978-11-14 | Med-El Inc. | Method and apparatus for automated classification and analysis of cells |
US4156570A (en) * | 1977-04-18 | 1979-05-29 | Robert A. Levine | Apparatus and method for measuring white blood cell and platelet concentrations in blood |
US4199748A (en) * | 1976-11-01 | 1980-04-22 | Rush-Presbyterian-St. Luke's Medical Center | Automated method and apparatus for classification of cells with application to the diagnosis of anemia |
US4209256A (en) * | 1978-07-21 | 1980-06-24 | Smithkline Corporation | Liquid flow viewing cell and method for analyzing liquid stream |
US4225229A (en) * | 1977-03-04 | 1980-09-30 | Goehde Wolfgang | Apparatus for measuring cytological properties |
EP0022670A2 (en) * | 1979-07-13 | 1981-01-21 | Ortho Diagnostic Systems Inc. | Method and apparatus for automated identification and enumeration of specified blood cell subclasses |
US4318886A (en) * | 1979-11-19 | 1982-03-09 | Nippon Kogaku K.K. | Automatic HLA typing apparatus |
US4336029A (en) * | 1980-08-15 | 1982-06-22 | Ortho Diagnostic Systems Inc. | Method and reagents for quantitative determination of reticulocytes and platelets in whole blood |
US4343782A (en) * | 1978-04-20 | 1982-08-10 | Shapiro Howard M | Cytological assay procedure |
US4354114A (en) * | 1979-10-09 | 1982-10-12 | Karnaukhov Valery N | Apparatus for investigation of fluorescence characteristics of microscopic objects |
GB2116712A (en) * | 1982-03-09 | 1983-09-28 | Lawrence Kass | Metachromatic dye sorption and fluorescent light emissive means for differential determination of developmental stages of neutrophilic granulocytic cells and other leukocytes |
US4420558A (en) * | 1981-02-12 | 1983-12-13 | Janssen Pharmaceutica N.V. | Bright field light microscopic method of enumerating and characterizing subtypes of white blood cells and their precursors |
DE3331017A1 (en) * | 1982-08-30 | 1984-03-08 | Becton, Dickinson and Co., 07652 Paramus, N.J. | METHOD AND APPARATUS FOR DIFFERENTIATING DIFFERENT CELL SUBPOPULATIONS |
EP0121262A2 (en) * | 1983-04-05 | 1984-10-10 | Becton Dickinson and Company | Method and apparatus for distinguishing multiple subpopulations of cells in a sample |
EP0132064A2 (en) * | 1983-07-18 | 1985-01-23 | Becton Dickinson and Company | Method for elimination of interference of selected cell populations in analytic cytology |
FR2551551A1 (en) * | 1983-09-02 | 1985-03-08 | Reanal Finomvegyszergyar | Reagent simultaneously determining thrombocyte and leucocyte concns. |
US4542518A (en) * | 1982-11-03 | 1985-09-17 | Anthony Thomas E | Electronic blood cell counter |
WO1985005640A1 (en) * | 1984-05-31 | 1985-12-19 | Coulter Electronics, Inc. | A reagent system and method for identification, enumeration and examination of classes and subclasses of blood leukocytes |
US4581223A (en) * | 1980-03-12 | 1986-04-08 | Lawrence Kass | Individual leukocyte determination by means of differential metachromatic dye sorption |
US4581334A (en) * | 1983-04-25 | 1986-04-08 | Ortho Diagnostics Systems, Inc. | Simultaneous detection of leukocyte phagocytic and killing ability |
US4595582A (en) * | 1981-11-20 | 1986-06-17 | Reanal Finomvegyszergyar | Dyestuff composition for histological examinations |
US4717655A (en) * | 1982-08-30 | 1988-01-05 | Becton, Dickinson And Company | Method and apparatus for distinguishing multiple subpopulations of cells |
EP0259834A2 (en) * | 1986-09-10 | 1988-03-16 | Toa Medical Electronics Co., Ltd. | Method of classifying leucocytes by flow cytometry |
EP0268766A2 (en) * | 1986-11-27 | 1988-06-01 | Toa Medical Electronics Co., Ltd. | Method of classifying leukocytes by flow cytometry and reagents used in the method |
WO1988005908A1 (en) * | 1987-02-04 | 1988-08-11 | Richmond Cell Screening Limited | Cell screening, apparatus and methods |
US4847910A (en) * | 1986-06-30 | 1989-07-11 | Hitachi, Ltd. | Automatic cell sample classifying apparatus |
EP0485945A2 (en) * | 1990-11-16 | 1992-05-20 | Toa Medical Electronics Co., Ltd. | Method of classifying leukocytes by flow cytometry |
US5123731A (en) * | 1988-02-01 | 1992-06-23 | Canon Kabushiki Kaisha | Particle measuring device |
EP0499693A2 (en) * | 1991-02-22 | 1992-08-26 | Toa Medical Electronics Co., Ltd. | Method of differentiating erythroblasts from other cells by flow cytometry |
US5175109A (en) * | 1986-09-10 | 1992-12-29 | Toa Medical Electronics Co., Ltd. | Reagent for classifying leukocytes by flow cytometry |
US5179026A (en) * | 1986-11-27 | 1993-01-12 | Toa Medical Electronics Co., Ltd. | Method of classifying leukocytes by flow cytometry and reagents used in the method |
US5187749A (en) * | 1990-04-10 | 1993-02-16 | Olympus Optical Co., Ltd. | Virus infection examination apparatus having automatic determination function and method therefor |
US5188935A (en) * | 1984-05-31 | 1993-02-23 | Coulter Electronics, Inc. | Reagent system and method for identification, enumeration and examination of classes and subclasses of blood leukocytes |
US5239360A (en) * | 1988-10-21 | 1993-08-24 | Applied Biosystems, Inc. | Lens for capillary electrophoresis and chromatography |
US5480804A (en) * | 1989-06-28 | 1996-01-02 | Kirin Beverage Corporation | Method of and apparatus for detecting microorganisms |
US5656501A (en) * | 1993-08-11 | 1997-08-12 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Flow cell device for monitoring blood or other cell suspension under flow |
EP0806664A2 (en) * | 1996-04-12 | 1997-11-12 | Toa Medical Electronics Co., Ltd. | A reagent for measuring reticulocytes and a method of measuring them |
WO1998020365A1 (en) * | 1996-11-05 | 1998-05-14 | Corrosion Consultants, Inc. | Ultraviolet light illumination and viewing system and method for fluorescent dye leak detection |
US5843790A (en) * | 1996-12-31 | 1998-12-01 | Ronn; Avigdor M. | Method for assaying photosensitizing drug in whole blood |
US5846834A (en) * | 1996-12-31 | 1998-12-08 | Ronn; Avigdor M. | Method for assaying photosensitizing drug in whole blood |
US5948686A (en) * | 1998-03-07 | 1999-09-07 | Robert A. Leuine | Method for performing blood cell counts |
WO1999046047A2 (en) * | 1998-03-10 | 1999-09-16 | Large Scale Proteomics Corporation | Detection and characterization of microorganisms |
US6122396A (en) * | 1996-12-16 | 2000-09-19 | Bio-Tech Imaging, Inc. | Method of and apparatus for automating detection of microorganisms |
WO2000079326A1 (en) * | 1999-06-18 | 2000-12-28 | Genomic Solutions Inc. | An automated, ccd-based microarray imaging system |
US20020028158A1 (en) * | 1998-03-07 | 2002-03-07 | Wardlaw Stephen C. | Apparatus for analyzing biologic fluids |
US20030001072A1 (en) * | 1999-10-07 | 2003-01-02 | Dorsel Andreas N. | Apparatus and method for autofocus |
US20030042428A1 (en) * | 2001-07-28 | 2003-03-06 | Berthold Technologies Gmbh & Co. Kg | Apparatus for measuring in particular luminescent and/or fluorescent radiation |
US20030226977A1 (en) * | 2002-06-11 | 2003-12-11 | Leica Microsystems Heidelberg Gmbh | Method for scanning microscopy, scanning microscope, and apparatus for coding an illuminating light beam |
US6723290B1 (en) | 1998-03-07 | 2004-04-20 | Levine Robert A | Container for holding biologic fluid for analysis |
US6779350B2 (en) | 2002-03-21 | 2004-08-24 | Ritchie Enginerring Company, Inc. | Compressor head, internal discriminator, external discriminator, manifold design for refrigerant recovery apparatus and vacuum sensor |
US6832491B2 (en) | 2002-03-21 | 2004-12-21 | Ritchie Engineering Company, Inc. | Compressor head, internal discriminator, external discriminator, manifold design for refrigerant recovery apparatus |
US20050070025A1 (en) * | 2003-08-07 | 2005-03-31 | Greg Mooradian | System and method incorporating ultraviolet spectral fluorescence technology in sensor applications |
US20060135861A1 (en) * | 2003-02-06 | 2006-06-22 | Koninklijke Philips Electronics N.V. | Apparatus and method for blood analysis |
US20080138852A1 (en) * | 2004-02-13 | 2008-06-12 | Winkelman James W | Identification of blood elements using inverted microscopy |
US20080212069A1 (en) * | 2007-01-26 | 2008-09-04 | Becton, Dickinson And Company | Method, system, and compositions for cell counting and analysis |
EP2267444A1 (en) * | 1998-03-07 | 2010-12-29 | Wardlaw Partners LP | Determination of white blood cell differential and reticulocyte counts |
US8797527B2 (en) | 2011-08-24 | 2014-08-05 | Abbott Point Of Care, Inc. | Biologic fluid sample analysis cartridge |
US20150029508A1 (en) * | 2013-07-23 | 2015-01-29 | Sysmex Corporation | Sample analyzing apparatus, disease monitoring system, and method for managing data of sample analyzing apparatus |
US9199233B2 (en) | 2010-03-31 | 2015-12-01 | Abbott Point Of Care, Inc. | Biologic fluid analysis cartridge with deflecting top panel |
US9523682B2 (en) | 2011-11-16 | 2016-12-20 | Becton, Dickinson And Company | Methods and systems for detecting an analyte in a sample |
US9579651B2 (en) | 2009-12-18 | 2017-02-28 | Abbott Point Of Care, Inc. | Biologic fluid analysis cartridge |
US9649061B2 (en) | 2015-03-10 | 2017-05-16 | Becton, Dickinson And Company | Biological fluid micro-sample management device |
US9678065B2 (en) | 2013-01-11 | 2017-06-13 | Becton, Dickinson And Company | Low-cost point-of-care assay device |
US9693723B2 (en) | 2014-10-14 | 2017-07-04 | Becton, Dickinson And Company | Blood sample management using open cell foam |
US9797899B2 (en) | 2013-11-06 | 2017-10-24 | Becton, Dickinson And Company | Microfluidic devices, and methods of making and using the same |
US9873118B2 (en) | 2010-12-30 | 2018-01-23 | Abbott Point Of Care, Inc. | Biologic fluid analysis cartridge with sample handling portion and analysis chamber portion |
US10018640B2 (en) | 2013-11-13 | 2018-07-10 | Becton, Dickinson And Company | Optical imaging system and methods for using the same |
US10371640B2 (en) | 2011-11-28 | 2019-08-06 | California Institute Of Technology | Compositions and methods for leukocyte differential counting |
US10488644B2 (en) | 2015-09-17 | 2019-11-26 | S.D. Sight Diagnostics Ltd. | Methods and apparatus for detecting an entity in a bodily sample |
CN110530838A (en) * | 2019-02-01 | 2019-12-03 | 东华大学 | A kind of multi-component dyes compatibility evaluation method |
US10578606B2 (en) | 2015-09-01 | 2020-03-03 | Becton, Dickinson And Company | Depth filtration device for separating specimen phases |
US10640807B2 (en) | 2011-12-29 | 2020-05-05 | S.D. Sight Diagnostics Ltd | Methods and systems for detecting a pathogen in a biological sample |
US10843190B2 (en) | 2010-12-29 | 2020-11-24 | S.D. Sight Diagnostics Ltd. | Apparatus and method for analyzing a bodily sample |
US11099175B2 (en) | 2016-05-11 | 2021-08-24 | S.D. Sight Diagnostics Ltd. | Performing optical measurements on a sample |
US11298061B2 (en) | 2014-10-14 | 2022-04-12 | Becton, Dickinson And Company | Blood sample management using open cell foam |
US11307196B2 (en) | 2016-05-11 | 2022-04-19 | S.D. Sight Diagnostics Ltd. | Sample carrier for optical measurements |
US11609413B2 (en) | 2017-11-14 | 2023-03-21 | S.D. Sight Diagnostics Ltd. | Sample carrier for microscopy and optical density measurements |
US11733150B2 (en) | 2016-03-30 | 2023-08-22 | S.D. Sight Diagnostics Ltd. | Distinguishing between blood sample components |
US11927519B2 (en) | 2015-02-09 | 2024-03-12 | Slingshot Biosciences, Inc. | Synthetic human cell mimic hydrogel particle for cytometric or coulter device |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3315229A (en) * | 1963-12-31 | 1967-04-18 | Ibm | Blood cell recognizer |
US3497690A (en) * | 1967-09-21 | 1970-02-24 | Bausch & Lomb | Method and apparatus for classifying biological cells by measuring the size and fluorescent response thereof |
-
1973
- 1973-05-31 US US365460A patent/US3916205A/en not_active Expired - Lifetime
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3315229A (en) * | 1963-12-31 | 1967-04-18 | Ibm | Blood cell recognizer |
US3497690A (en) * | 1967-09-21 | 1970-02-24 | Bausch & Lomb | Method and apparatus for classifying biological cells by measuring the size and fluorescent response thereof |
Cited By (159)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4125828A (en) * | 1972-08-04 | 1978-11-14 | Med-El Inc. | Method and apparatus for automated classification and analysis of cells |
US3992631A (en) * | 1975-02-27 | 1976-11-16 | International Diagnostic Technology, Inc. | Fluorometric system, method and test article |
US4000417A (en) * | 1975-08-25 | 1976-12-28 | Honeywell Inc. | Scanning microscope system with automatic cell find and autofocus |
US4015595A (en) * | 1975-09-15 | 1977-04-05 | Benjamin Jr J Malvern | Photoplethysmographs |
US4071891A (en) * | 1976-04-30 | 1978-01-31 | Barrows George H | Electronic calculator - register for hematology differentials |
US4025310A (en) * | 1976-05-28 | 1977-05-24 | International Diagnostic Technology, Inc. | Method for reading a wet fluorescent surface |
US4199748A (en) * | 1976-11-01 | 1980-04-22 | Rush-Presbyterian-St. Luke's Medical Center | Automated method and apparatus for classification of cells with application to the diagnosis of anemia |
US4097845A (en) * | 1976-11-01 | 1978-06-27 | Rush-Presbyterian-St. Luke's Medical Center | Method of and an apparatus for automatic classification of red blood cells |
US4225229A (en) * | 1977-03-04 | 1980-09-30 | Goehde Wolfgang | Apparatus for measuring cytological properties |
US4156570A (en) * | 1977-04-18 | 1979-05-29 | Robert A. Levine | Apparatus and method for measuring white blood cell and platelet concentrations in blood |
US4117338A (en) * | 1977-05-24 | 1978-09-26 | Corning Glass Works | Automatic recording fluorometer/densitometer |
US4343782A (en) * | 1978-04-20 | 1982-08-10 | Shapiro Howard M | Cytological assay procedure |
US4209256A (en) * | 1978-07-21 | 1980-06-24 | Smithkline Corporation | Liquid flow viewing cell and method for analyzing liquid stream |
EP0022670A2 (en) * | 1979-07-13 | 1981-01-21 | Ortho Diagnostic Systems Inc. | Method and apparatus for automated identification and enumeration of specified blood cell subclasses |
US4284412A (en) * | 1979-07-13 | 1981-08-18 | Ortho Diagnostics, Inc. | Method and apparatus for automated identification and enumeration of specified blood cell subclasses |
EP0022670A3 (en) * | 1979-07-13 | 1981-05-20 | Ortho Diagnostic Systems Inc. | Method and apparatus for automated identification and enumeration of specified blood cell subclasses |
US4354114A (en) * | 1979-10-09 | 1982-10-12 | Karnaukhov Valery N | Apparatus for investigation of fluorescence characteristics of microscopic objects |
US4318886A (en) * | 1979-11-19 | 1982-03-09 | Nippon Kogaku K.K. | Automatic HLA typing apparatus |
US4581223A (en) * | 1980-03-12 | 1986-04-08 | Lawrence Kass | Individual leukocyte determination by means of differential metachromatic dye sorption |
US4336029A (en) * | 1980-08-15 | 1982-06-22 | Ortho Diagnostic Systems Inc. | Method and reagents for quantitative determination of reticulocytes and platelets in whole blood |
US4420558A (en) * | 1981-02-12 | 1983-12-13 | Janssen Pharmaceutica N.V. | Bright field light microscopic method of enumerating and characterizing subtypes of white blood cells and their precursors |
US4500509A (en) * | 1981-03-11 | 1985-02-19 | Lawrence Kass | Metachromatic dye sorption and fluorescent light emmisive means for differential determination of developmental stages of neutrophilic granulocytic cells and other leukocytes |
US4595582A (en) * | 1981-11-20 | 1986-06-17 | Reanal Finomvegyszergyar | Dyestuff composition for histological examinations |
GB2116712A (en) * | 1982-03-09 | 1983-09-28 | Lawrence Kass | Metachromatic dye sorption and fluorescent light emissive means for differential determination of developmental stages of neutrophilic granulocytic cells and other leukocytes |
US4717655A (en) * | 1982-08-30 | 1988-01-05 | Becton, Dickinson And Company | Method and apparatus for distinguishing multiple subpopulations of cells |
US4499052A (en) * | 1982-08-30 | 1985-02-12 | Becton, Dickinson And Company | Apparatus for distinguishing multiple subpopulations of cells |
DE3331017A1 (en) * | 1982-08-30 | 1984-03-08 | Becton, Dickinson and Co., 07652 Paramus, N.J. | METHOD AND APPARATUS FOR DIFFERENTIATING DIFFERENT CELL SUBPOPULATIONS |
JPH0447265B2 (en) * | 1982-08-30 | 1992-08-03 | Becton Dickinson Co | |
JPS5960261A (en) * | 1982-08-30 | 1984-04-06 | ベクトン・デイツキンソン・アンド・カンパニ− | Method of discriminating large number of preliminary prolif-eration of cell |
US4542518A (en) * | 1982-11-03 | 1985-09-17 | Anthony Thomas E | Electronic blood cell counter |
EP0121262A3 (en) * | 1983-04-05 | 1985-11-06 | Becton, Dickinson And Company | Method and apparatus for distinguishing multiple subpopulations of cells in a sample |
JPS59184862A (en) * | 1983-04-05 | 1984-10-20 | ベクトン・ディッキンソン・アンド・カンパニ− | Method and device for discriminating plurality of subsidiarygroup of cell in sample |
JPH058385B2 (en) * | 1983-04-05 | 1993-02-02 | Becton Dickinson Co | |
EP0121262A2 (en) * | 1983-04-05 | 1984-10-10 | Becton Dickinson and Company | Method and apparatus for distinguishing multiple subpopulations of cells in a sample |
US4581334A (en) * | 1983-04-25 | 1986-04-08 | Ortho Diagnostics Systems, Inc. | Simultaneous detection of leukocyte phagocytic and killing ability |
EP0132064A3 (en) * | 1983-07-18 | 1988-04-06 | Becton, Dickinson And Company | Method for elimination of selected cell populations in analytic cytology |
JPH058386B2 (en) * | 1983-07-18 | 1993-02-02 | Becton Dickinson Co | |
EP0132064A2 (en) * | 1983-07-18 | 1985-01-23 | Becton Dickinson and Company | Method for elimination of interference of selected cell populations in analytic cytology |
JPS6076666A (en) * | 1983-07-18 | 1985-05-01 | ベクトン・デイツキンソン・アンド・カンパニ− | Method of removing specific cell distribution in cytology for analysis |
US4599307A (en) * | 1983-07-18 | 1986-07-08 | Becton, Dickinson And Company | Method for elimination of selected cell populations in analytic cytology |
FR2551551A1 (en) * | 1983-09-02 | 1985-03-08 | Reanal Finomvegyszergyar | Reagent simultaneously determining thrombocyte and leucocyte concns. |
US5188935A (en) * | 1984-05-31 | 1993-02-23 | Coulter Electronics, Inc. | Reagent system and method for identification, enumeration and examination of classes and subclasses of blood leukocytes |
WO1985005640A1 (en) * | 1984-05-31 | 1985-12-19 | Coulter Electronics, Inc. | A reagent system and method for identification, enumeration and examination of classes and subclasses of blood leukocytes |
US4847910A (en) * | 1986-06-30 | 1989-07-11 | Hitachi, Ltd. | Automatic cell sample classifying apparatus |
US5296378A (en) * | 1986-09-10 | 1994-03-22 | Toa Medical Electronics Co., Ltd. | Method for classifying leukocytes by flow cytometry |
EP0259834A3 (en) * | 1986-09-10 | 1988-12-07 | Toa Medical Electronics Co., Ltd. | Method of classifying leucocytes by flow cytometry |
US4933293A (en) * | 1986-09-10 | 1990-06-12 | Toa Medical Electronics Co., Ltd. | Method of classifying leukocytes by flow cytometry and reagents used in the method |
US5928949A (en) * | 1986-09-10 | 1999-07-27 | Toa Medical Electronics Co., Ltd. | Reagent and method for classifying leukocytes by flow cytometry |
EP0259834A2 (en) * | 1986-09-10 | 1988-03-16 | Toa Medical Electronics Co., Ltd. | Method of classifying leucocytes by flow cytometry |
US5175109A (en) * | 1986-09-10 | 1992-12-29 | Toa Medical Electronics Co., Ltd. | Reagent for classifying leukocytes by flow cytometry |
EP0268766A3 (en) * | 1986-11-27 | 1988-12-14 | Toa Medical Electronics Co., Ltd. | Method of classifying leukocytes by flow cytometry and reagents used in the method |
US5179026A (en) * | 1986-11-27 | 1993-01-12 | Toa Medical Electronics Co., Ltd. | Method of classifying leukocytes by flow cytometry and reagents used in the method |
US5039613A (en) * | 1986-11-27 | 1991-08-13 | Toa Medical Electronics Co., Ltd. | Reagents used in a method of classifying leukocytes by flow cytometry |
EP0268766A2 (en) * | 1986-11-27 | 1988-06-01 | Toa Medical Electronics Co., Ltd. | Method of classifying leukocytes by flow cytometry and reagents used in the method |
GB2209395A (en) * | 1987-02-04 | 1989-05-10 | Richmond Cell Screening Ltd | Cell screening, apparatus and methods |
WO1988005908A1 (en) * | 1987-02-04 | 1988-08-11 | Richmond Cell Screening Limited | Cell screening, apparatus and methods |
GB2209395B (en) * | 1987-02-04 | 1991-09-04 | Richmond Cell Screening Ltd | Cell screening, apparatus and methods |
US5123731A (en) * | 1988-02-01 | 1992-06-23 | Canon Kabushiki Kaisha | Particle measuring device |
US5239360A (en) * | 1988-10-21 | 1993-08-24 | Applied Biosystems, Inc. | Lens for capillary electrophoresis and chromatography |
US5480804A (en) * | 1989-06-28 | 1996-01-02 | Kirin Beverage Corporation | Method of and apparatus for detecting microorganisms |
US5187749A (en) * | 1990-04-10 | 1993-02-16 | Olympus Optical Co., Ltd. | Virus infection examination apparatus having automatic determination function and method therefor |
EP0485945A3 (en) * | 1990-11-16 | 1993-02-03 | Toa Medical Electronics | Method of classifying leukocytes by flow cytometry |
EP0485945A2 (en) * | 1990-11-16 | 1992-05-20 | Toa Medical Electronics Co., Ltd. | Method of classifying leukocytes by flow cytometry |
EP0499693A3 (en) * | 1991-02-22 | 1993-02-03 | Toa Medical Electronics Co., Ltd. | Method of differentiating erythroblasts from other cells by flow cytometry |
US5298426A (en) * | 1991-02-22 | 1994-03-29 | Toa Medical Electronics Co., Ltd. | Method of differentiating erythroblasts from other cells by flow cytometry |
EP0499693A2 (en) * | 1991-02-22 | 1992-08-26 | Toa Medical Electronics Co., Ltd. | Method of differentiating erythroblasts from other cells by flow cytometry |
US5656501A (en) * | 1993-08-11 | 1997-08-12 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Flow cell device for monitoring blood or other cell suspension under flow |
EP0806664A3 (en) * | 1996-04-12 | 1998-04-08 | Toa Medical Electronics Co., Ltd. | A reagent for measuring reticulocytes and a method of measuring them |
EP0806664A2 (en) * | 1996-04-12 | 1997-11-12 | Toa Medical Electronics Co., Ltd. | A reagent for measuring reticulocytes and a method of measuring them |
WO1998020365A1 (en) * | 1996-11-05 | 1998-05-14 | Corrosion Consultants, Inc. | Ultraviolet light illumination and viewing system and method for fluorescent dye leak detection |
US6122396A (en) * | 1996-12-16 | 2000-09-19 | Bio-Tech Imaging, Inc. | Method of and apparatus for automating detection of microorganisms |
US5843790A (en) * | 1996-12-31 | 1998-12-01 | Ronn; Avigdor M. | Method for assaying photosensitizing drug in whole blood |
US5846834A (en) * | 1996-12-31 | 1998-12-08 | Ronn; Avigdor M. | Method for assaying photosensitizing drug in whole blood |
EP2267443A1 (en) * | 1998-03-07 | 2010-12-29 | Wardlaw Partners LP | Determination of white blood cell differential and reticulocyte counts |
EP2320228A1 (en) * | 1998-03-07 | 2011-05-11 | Wardlaw Partners LP | Determination of white blood cell differential and reticulocyte counts |
US8367012B2 (en) | 1998-03-07 | 2013-02-05 | Abbott Laboratories | Container for holding biologic fluid for analysis |
US5948686A (en) * | 1998-03-07 | 1999-09-07 | Robert A. Leuine | Method for performing blood cell counts |
EP2267444A1 (en) * | 1998-03-07 | 2010-12-29 | Wardlaw Partners LP | Determination of white blood cell differential and reticulocyte counts |
US6723290B1 (en) | 1998-03-07 | 2004-04-20 | Levine Robert A | Container for holding biologic fluid for analysis |
US20080187466A1 (en) * | 1998-03-07 | 2008-08-07 | Wardlaw Stephen C | Container for holding biologic fluid for analysis |
US20020028158A1 (en) * | 1998-03-07 | 2002-03-07 | Wardlaw Stephen C. | Apparatus for analyzing biologic fluids |
US6929953B1 (en) | 1998-03-07 | 2005-08-16 | Robert A. Levine | Apparatus for analyzing biologic fluids |
US6869570B2 (en) | 1998-03-07 | 2005-03-22 | Robert A. Levine | Apparatus for analyzing biologic fluids |
US6866823B2 (en) | 1998-03-07 | 2005-03-15 | Robert A. Levine | Apparatus for analyzing biologic fluids |
US20040156755A1 (en) * | 1998-03-07 | 2004-08-12 | Robert Levine | Container for holding biologic fluid for analysis |
US6340570B1 (en) | 1998-03-10 | 2002-01-22 | Large Scale Proteomics Corp. | Detection and characterization of microorganisms |
US6911312B2 (en) | 1998-03-10 | 2005-06-28 | Large Scale Proteomics Corporation | Detection and characterization of microorganisms |
US7070739B1 (en) | 1998-03-10 | 2006-07-04 | Large Scale Proteomics Corporation | Detection and characterization of microorganisms |
WO1999046047A2 (en) * | 1998-03-10 | 1999-09-16 | Large Scale Proteomics Corporation | Detection and characterization of microorganisms |
US20020127546A1 (en) * | 1998-03-10 | 2002-09-12 | Anderson Norman G. | Detection and characterization of microorganisms |
US6346421B1 (en) | 1998-03-10 | 2002-02-12 | Large Scale Proteomics Corp. | Methods for concentrating and detecting microorganisms using centrifuge tubes |
US6254834B1 (en) | 1998-03-10 | 2001-07-03 | Large Scale Proteomics Corp. | Detection and characterization of microorganisms |
US6479239B1 (en) | 1998-03-10 | 2002-11-12 | Large Scale Biology Corporation | Detection and characterization of microorganisms |
US20020137026A1 (en) * | 1998-03-10 | 2002-09-26 | Anderson Norman G. | Detection and characterization of microorganisms |
WO1999046047A3 (en) * | 1998-03-10 | 1999-12-02 | Biosource Proteomics Inc | Detection and characterization of microorganisms |
WO2000079326A1 (en) * | 1999-06-18 | 2000-12-28 | Genomic Solutions Inc. | An automated, ccd-based microarray imaging system |
US6496309B1 (en) * | 1999-06-18 | 2002-12-17 | Genomic Solutions, Inc. | Automated, CCD-based DNA micro-array imaging system |
US20030001072A1 (en) * | 1999-10-07 | 2003-01-02 | Dorsel Andreas N. | Apparatus and method for autofocus |
US6949754B2 (en) * | 2001-07-28 | 2005-09-27 | Berthold Technologies Gmbh & Co. Kg | Apparatus for measuring in particular luminescent and/or fluorescent radiation |
US20030042428A1 (en) * | 2001-07-28 | 2003-03-06 | Berthold Technologies Gmbh & Co. Kg | Apparatus for measuring in particular luminescent and/or fluorescent radiation |
US6832491B2 (en) | 2002-03-21 | 2004-12-21 | Ritchie Engineering Company, Inc. | Compressor head, internal discriminator, external discriminator, manifold design for refrigerant recovery apparatus |
US7159412B2 (en) | 2002-03-21 | 2007-01-09 | Ritchie Engineering Company, Inc. | Compressor head, internal discriminator, external discriminator, manifold design for refrigeration recovery apparatus |
US7310965B2 (en) | 2002-03-21 | 2007-12-25 | Ritchie Engineering Company, Inc. | Compressor head, internal discriminator, external discriminator, manifold design for refrigeration recovery apparatus |
US7073346B2 (en) | 2002-03-21 | 2006-07-11 | Ritchie Engineering Company, Inc. | Compressor head, internal discriminator, external discriminator, manifold design for refrigerant recovery apparatus and vacuum sensor |
US6779350B2 (en) | 2002-03-21 | 2004-08-24 | Ritchie Enginerring Company, Inc. | Compressor head, internal discriminator, external discriminator, manifold design for refrigerant recovery apparatus and vacuum sensor |
US7133130B2 (en) * | 2002-06-11 | 2006-11-07 | Leica Microsystems Cms Gmbh | Method for scanning microscopy, scanning microscope, and apparatus for coding an illuminating light beam |
US20030226977A1 (en) * | 2002-06-11 | 2003-12-11 | Leica Microsystems Heidelberg Gmbh | Method for scanning microscopy, scanning microscope, and apparatus for coding an illuminating light beam |
US20060135861A1 (en) * | 2003-02-06 | 2006-06-22 | Koninklijke Philips Electronics N.V. | Apparatus and method for blood analysis |
US20050070025A1 (en) * | 2003-08-07 | 2005-03-31 | Greg Mooradian | System and method incorporating ultraviolet spectral fluorescence technology in sensor applications |
US9176121B2 (en) * | 2004-02-13 | 2015-11-03 | Roche Diagnostics Hematology, Inc. | Identification of blood elements using inverted microscopy |
US20080138852A1 (en) * | 2004-02-13 | 2008-06-12 | Winkelman James W | Identification of blood elements using inverted microscopy |
US20080212069A1 (en) * | 2007-01-26 | 2008-09-04 | Becton, Dickinson And Company | Method, system, and compositions for cell counting and analysis |
US9097640B2 (en) | 2007-01-26 | 2015-08-04 | Becton, Dickinson And Company | Method, system, and compositions for cell counting and analysis |
US7738094B2 (en) | 2007-01-26 | 2010-06-15 | Becton, Dickinson And Company | Method, system, and compositions for cell counting and analysis |
US9579651B2 (en) | 2009-12-18 | 2017-02-28 | Abbott Point Of Care, Inc. | Biologic fluid analysis cartridge |
US9993817B2 (en) | 2009-12-18 | 2018-06-12 | Abbott Point Of Care, Inc. | Biologic fluid analysis cartridge |
US9199233B2 (en) | 2010-03-31 | 2015-12-01 | Abbott Point Of Care, Inc. | Biologic fluid analysis cartridge with deflecting top panel |
US10843190B2 (en) | 2010-12-29 | 2020-11-24 | S.D. Sight Diagnostics Ltd. | Apparatus and method for analyzing a bodily sample |
US11583851B2 (en) | 2010-12-30 | 2023-02-21 | Abbott Point Of Care Inc. | Biologic fluid analysis cartridge with sample handling portion and analysis chamber portion |
US9873118B2 (en) | 2010-12-30 | 2018-01-23 | Abbott Point Of Care, Inc. | Biologic fluid analysis cartridge with sample handling portion and analysis chamber portion |
US10391487B2 (en) | 2010-12-30 | 2019-08-27 | Abbott Point Of Care, Inc. | Biologic fluid analysis cartridge with sample handling portion and analysis chamber portion |
US8797527B2 (en) | 2011-08-24 | 2014-08-05 | Abbott Point Of Care, Inc. | Biologic fluid sample analysis cartridge |
US9523682B2 (en) | 2011-11-16 | 2016-12-20 | Becton, Dickinson And Company | Methods and systems for detecting an analyte in a sample |
US10371640B2 (en) | 2011-11-28 | 2019-08-06 | California Institute Of Technology | Compositions and methods for leukocyte differential counting |
US11584950B2 (en) | 2011-12-29 | 2023-02-21 | S.D. Sight Diagnostics Ltd. | Methods and systems for detecting entities in a biological sample |
US10640807B2 (en) | 2011-12-29 | 2020-05-05 | S.D. Sight Diagnostics Ltd | Methods and systems for detecting a pathogen in a biological sample |
US9678065B2 (en) | 2013-01-11 | 2017-06-13 | Becton, Dickinson And Company | Low-cost point-of-care assay device |
US20150029508A1 (en) * | 2013-07-23 | 2015-01-29 | Sysmex Corporation | Sample analyzing apparatus, disease monitoring system, and method for managing data of sample analyzing apparatus |
US10073080B2 (en) * | 2013-07-23 | 2018-09-11 | Sysmex Corporation | Sample analyzing apparatus, disease monitoring system, and method for managing multiple disease determination data in a sample analyzing apparatus |
US9797899B2 (en) | 2013-11-06 | 2017-10-24 | Becton, Dickinson And Company | Microfluidic devices, and methods of making and using the same |
US10073093B2 (en) | 2013-11-06 | 2018-09-11 | Becton, Dickinson And Company | Microfluidic devices, and methods of making and using the same |
US10018640B2 (en) | 2013-11-13 | 2018-07-10 | Becton, Dickinson And Company | Optical imaging system and methods for using the same |
US10663476B2 (en) | 2013-11-13 | 2020-05-26 | Becton, Dickinson And Company | Optical imaging system and methods for using the same |
US10888261B2 (en) | 2014-10-14 | 2021-01-12 | Becton, Dickinson And Company | Blood sample management using open cell foam |
US10219731B2 (en) | 2014-10-14 | 2019-03-05 | Becton, Dickinson And Company | Blood sample management using open cell foam |
US11134875B2 (en) | 2014-10-14 | 2021-10-05 | Becton, Dickinson And Company | Blood sample management using open cell foam |
US10595762B2 (en) | 2014-10-14 | 2020-03-24 | Becton, Dickinson And Company | Blood sample management using open cell foam |
US9693723B2 (en) | 2014-10-14 | 2017-07-04 | Becton, Dickinson And Company | Blood sample management using open cell foam |
US11298061B2 (en) | 2014-10-14 | 2022-04-12 | Becton, Dickinson And Company | Blood sample management using open cell foam |
US11927519B2 (en) | 2015-02-09 | 2024-03-12 | Slingshot Biosciences, Inc. | Synthetic human cell mimic hydrogel particle for cytometric or coulter device |
US9873117B2 (en) | 2015-03-10 | 2018-01-23 | Becton, Dickinson And Company | Biological fluid micro-sample management device |
US9649061B2 (en) | 2015-03-10 | 2017-05-16 | Becton, Dickinson And Company | Biological fluid micro-sample management device |
US10578606B2 (en) | 2015-09-01 | 2020-03-03 | Becton, Dickinson And Company | Depth filtration device for separating specimen phases |
US11366095B2 (en) | 2015-09-01 | 2022-06-21 | Becton, Dickinson And Company | Depth filtration device for separating specimen phases |
US11808757B2 (en) | 2015-09-01 | 2023-11-07 | Becton, Dickinson And Company | Depth filtration device for separating specimen phases |
US11262571B2 (en) | 2015-09-17 | 2022-03-01 | S.D. Sight Diagnostics Ltd. | Determining a staining-quality parameter of a blood sample |
US11796788B2 (en) | 2015-09-17 | 2023-10-24 | S.D. Sight Diagnostics Ltd. | Detecting a defect within a bodily sample |
US11199690B2 (en) | 2015-09-17 | 2021-12-14 | S.D. Sight Diagnostics Ltd. | Determining a degree of red blood cell deformity within a blood sample |
US10488644B2 (en) | 2015-09-17 | 2019-11-26 | S.D. Sight Diagnostics Ltd. | Methods and apparatus for detecting an entity in a bodily sample |
US10663712B2 (en) | 2015-09-17 | 2020-05-26 | S.D. Sight Diagnostics Ltd. | Methods and apparatus for detecting an entity in a bodily sample |
US11914133B2 (en) | 2015-09-17 | 2024-02-27 | S.D. Sight Diagnostics Ltd. | Methods and apparatus for analyzing a bodily sample |
US11733150B2 (en) | 2016-03-30 | 2023-08-22 | S.D. Sight Diagnostics Ltd. | Distinguishing between blood sample components |
US11307196B2 (en) | 2016-05-11 | 2022-04-19 | S.D. Sight Diagnostics Ltd. | Sample carrier for optical measurements |
US11808758B2 (en) | 2016-05-11 | 2023-11-07 | S.D. Sight Diagnostics Ltd. | Sample carrier for optical measurements |
US11099175B2 (en) | 2016-05-11 | 2021-08-24 | S.D. Sight Diagnostics Ltd. | Performing optical measurements on a sample |
US11614609B2 (en) | 2017-11-14 | 2023-03-28 | S.D. Sight Diagnostics Ltd. | Sample carrier for microscopy measurements |
US11609413B2 (en) | 2017-11-14 | 2023-03-21 | S.D. Sight Diagnostics Ltd. | Sample carrier for microscopy and optical density measurements |
US11921272B2 (en) | 2017-11-14 | 2024-03-05 | S.D. Sight Diagnostics Ltd. | Sample carrier for optical measurements |
CN110530838A (en) * | 2019-02-01 | 2019-12-03 | 东华大学 | A kind of multi-component dyes compatibility evaluation method |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US3916205A (en) | Differential counting of leukocytes and other cells | |
US4146604A (en) | Differential counting of leukocytes and other cells | |
US4933293A (en) | Method of classifying leukocytes by flow cytometry and reagents used in the method | |
Vonesch et al. | The colored revolution of bioimaging | |
US3918812A (en) | Diagnoses of disease states by fluorescent measurements utilizing scanning laser beams | |
US5528045A (en) | Particle analyzer with spatially split wavelength filter | |
US5175109A (en) | Reagent for classifying leukocytes by flow cytometry | |
Rost | Quantitative fluorescence microscopy | |
CA1309328C (en) | Method of classifying leukocytes by flow cytometry and reagents used in the method | |
EP0160568B1 (en) | Methods and apparatus for analysis of particles and cells | |
US5428441A (en) | Apparatus for analyzing particle images | |
JPS63118638A (en) | Method and device for determining characteristic of cell, particle, etc. by using multiple fluorometric analysis | |
JPH04337459A (en) | Reagent and method for analyzing cell in urine | |
JPS63500334A (en) | Scattered light measurement device by biological cells for flow cytometer | |
JP3815838B2 (en) | Particle measuring device | |
Picciolo et al. | Reduction of fading of fluorescent reaction product for microphotometric quantitation | |
US4777133A (en) | Device for quantitative endpoint determination in immunofluorescence using microfluorophotometry | |
US4622291A (en) | Method and device for quantitative end point determination in immunofluorescence using microfluorophotometry | |
Tanke et al. | A parameter for the distribution of fluorophores in cells derived from measurements of inner filter effect and reabsorption phenomenon | |
Berns | Fluorescence analysis of cells using a laser light source | |
Ploem | Appropriate technology for the quantitative assessment of the final reaction product of histochemical techniques | |
HU231357B1 (en) | Method and apparatus for analysing fluorescence of a sample | |
Jotz et al. | A new optical multichannel microspectrofluorometer. | |
Wittrup et al. | Fluorescence array detector for large‐field quantitative fluorescence cytometry | |
Koper et al. | An epiilluminator/detector unit permitting arc lamp illumination for fluorescence activated cell sorters |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: BIO-RAD LABORATORIES, INC., A CORP. OF DE. Free format text: MERGER;ASSIGNOR:BLOCK ENGINEERING, INC.;REEL/FRAME:003974/0501 Effective date: 19820406 |
|
AS | Assignment |
Owner name: FIRST NATIONAL BANK OF BOSTON, THE Free format text: SECURITY INTEREST;ASSIGNOR:ORION RESEARCH INCORPORATED, A MA. CORP.;REEL/FRAME:004666/0862 Effective date: 19860624 |
|
STCF | Information on status: patent grant |
Free format text: PATENTED FILE - (OLD CASE ADDED FOR FILE TRACKING PURPOSES) |
|
AS | Assignment |
Owner name: ORION RESEARCH INCORPORATED, A CORP. OF MA Free format text: RELEASED BY SECURED PARTY;ASSIGNOR:FIRST NATIONAL BANK OF BOSTON, THE;REEL/FRAME:005688/0306 Effective date: 19910430 |
|
AS | Assignment |
Owner name: FIRST NATIONAL BANK OF BOSTON, THE A NATIONAL BAN Free format text: SECURITY INTEREST;ASSIGNOR:ORION RESEARCH INCORPORATED A CORPORATION OF MASSACHUSETTS;REEL/FRAME:005709/0678 Effective date: 19910502 |
|
AS | Assignment |
Owner name: ORION RESEARCH, INC., MASSACHUSETTS Free format text: RELEASE BY SECURED PARTY;ASSIGNOR:FIRST NATIONAL BANK OF BOSTON, THE;REEL/FRAME:006554/0561 Effective date: 19930520 |