US4146604A
Expired / Lapsed
US
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Differential counting of leukocytes and other cells
Assignee: Bio Rad Laboratories Inc
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Filing: 1974-01-30
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Granted: 1979-03-27
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Expires: 1994-01-30
Abstract
Abstract A novel composition of matter, process and apparatus is provided for distinguishing and absolutely counting leukocytes, erythrocytes, reticulocytes, and other cells as well as differently counting and classifying the different types of leukocytes. The composition of matter comprises a plurality of compatible dyes for uniquely distinguishing leukocytes, erythrocytes, reticulocytes, and other cells randomly distributed in a medium. At least one of the dyes imparts a characteristic fluorescence to all leukocytes stained therewith. The nature of the dye composition is such that when the stained leukocytes are irradiated with radiation having the characteristic absorption wavelengths of each dye, the relative intensities of the light emitted and/or transmitted by each leukocyte type defines that particular leukocyte type. Apparatus is therefore provided for irradiating the medium with the characteristic radiation and measuring the radiation emitted by the irradiated leukocytes in each of their characteristic wavelength regions.
Inventors
- 👤 Marcos Y. Kleinerman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 22:09
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/5094; G01N15/00; G01N15/10; G01N15/14; G01N15/1468; G01N15/147; G01N21/00; G01N21/62; G01N21/63; G01N21/64; G01N21/6428; G01N15/01; G01N2015/016; G01N2021/6439; G01N2021/6441; Y; Y10; Y10S; Y10S436/00; Y10S436/80
Claims Analysis
Total Claims: 31
Claim 1
Independent Claim
89 words
A mixture for staining a biological cell and comprising, in combination; a fluorescent dye capable of dyeing a first specific constituent of said cell, said dye being a sulfonated triazinyl derivative of diamino stilbene, and a sensitizer compound capable of being coupled to a different specific constituent of said cell and capable of absorbing incident radiation in a spectral absorption band and of transmitting to dye disposed adjacent to said compound, at least part of the energy of the absorbed incident radiation to excite said dye into fluorescent emission.
Claim 2
Dependent Claim
89 words
A mixture for staining a biological cell and comprising, in combination; a fluorescent dye capable of dyeing a first specific constituent of said cell, said dye being a sulfonated triazinyl derivative of diamino stilbene, and a sensitizer compound capable of being coupled to a different specific constituent of said cell and capable of absorbing incident radiation in a spectral absorption band and of transmitting to dye disposed adjacent to said compound, at least part of the energy of the absorbed incident radiation to excite said dye into fluorescent emission.
Claim 3
Dependent Claim
73 words
A dye mixture for distinguishing among biological cell types, said mixture comprising a plurality of compatible dyes, which dyes differentially stain characteristic structures of the cells types of interest, one of said dyes being a sulfonated triazinyl derivative of diamino stilbene, which differentially fluorochrome stains protein structures of the cell types of interest, another one of said dyes being a cationic dye which differentially stains nucleic acids in the cell types of interest.
Claim 4
Dependent Claim
73 words
A dye mixture for distinguishing among biological cell types, said mixture comprising a plurality of compatible dyes, which dyes differentially stain characteristic structures of the cells types of interest, one of said dyes being a sulfonated triazinyl derivative of diamino stilbene, which differentially fluorochrome stains protein structures of the cell types of interest, another one of said dyes being a cationic dye which differentially stains nucleic acids in the cell types of interest.
Claim 5
Dependent Claim
15 words
The mixture as defined in claim 2 wherein said cationic dye is a phenanthridium dye.
Claim 6
Dependent Claim
14 words
The mixture as defined in claim 3 wherein said phenanthridium dye is ethidium bromide.
Claim 7
Dependent Claim
30 words
The mixture as defined in claim 2 wherein said sulfonated triazinyl derivative of diamino stilbene is 4,4' bis[4-(3 sulfoanilino)-6-[bis (2-hydroxy ethyl) amino]-1,3,5 triazin-2yl] amino stilbene 2,2' disulfonic acid tetrasodium salt.
Claim 8
Dependent Claim
15 words
The mixture as defined in claim 5 wherein said cationic dye is a phenanthridium dye.
Claim 9
Dependent Claim
14 words
The mixture as defined in claim 6 wherein said phenanthridium dye is ethidium bromide.
Claim 10
Dependent Claim
78 words
A dye mixture for distinguishing among blood cells, said mixture comprising a plurality of compatible dyes, which dyes differentially stain characteristic structures of the cells types of interest, one of said dyes being a sulfonated triazinyl derivative of diamino stilbene, which differentially fluorochrome stains the cytoplasm of leukocytes and the granules of granulocytes, another one of said dyes being a cationic dye which differentially stains nucleic acids in said blood cells and the filaments network structure of reticulocytes.
Claim 11
Dependent Claim
78 words
A dye mixture for distinguishing among blood cells, said mixture comprising a plurality of compatible dyes, which dyes differentially stain characteristic structures of the cells types of interest, one of said dyes being a sulfonated triazinyl derivative of diamino stilbene, which differentially fluorochrome stains the cytoplasm of leukocytes and the granules of granulocytes, another one of said dyes being a cationic dye which differentially stains nucleic acids in said blood cells and the filaments network structure of reticulocytes.
Claim 12
Dependent Claim
15 words
The mixture as defined in claim 8 wherein said cationic dye is a phenanthridium dye.
Claim 13
Dependent Claim
14 words
The mixture as defined in claim 9 wherein said phenanthridium dye is ethidium bromide.
Claim 14
Dependent Claim
29 words
The mixture as defined in claim 8 in which another one of said dyes is a sulfonated fluorescent derivative of 1,8 naphthalimide which differentially stains the granules of eosinophils.
Claim 15
Dependent Claim
18 words
The mixture as defined in claim 11 wherein said sulfonated fluorescent derivative of 1,8 naphthalimide is brilliant sulfaflavine.
Claim 16
Dependent Claim
29 words
The mixture as defined in claim 8 in which another one of said dyes is a toluidino derivative of naphthalene sulfonic acid which differentially stains the granules of eosinophils.
Claim 17
Dependent Claim
19 words
The mixture as defined in claim 13 wherein said toluidino derivative of naphthalene sulfonic acid is 8-p-toluidino-1-naphtheline sulfonic acid.
Claim 18
Dependent Claim
32 words
A dye mixture for differentially staining biological cells said mixture comprising: (a) a sulfonated triazinyl derivative of diamino stilbene; (b) a cationic dye; and (c) a sulfonated fluorescent derivative of 1,8 naphthalimide.
Claim 19
Dependent Claim
32 words
A dye mixture for differentially staining biological cells said mixture comprising: (a) a sulfonated triazinyl derivative of diamino stilbene; (b) a cationic dye; and (c) a sulfonated fluorescent derivative of 1,8 naphthalimide.
Claim 20
Dependent Claim
30 words
The mixture as defined in claim 15 wherein said sulfonated triazinyl derivative of diamino stilbene is 4,4' bis [4-(3 sulfoanilino)-6-[bis (2-hydroxy-ethyl) amino]1,3,5 triazin-2yl] amino stilbene 2,2' disulfonic acid tetrasodium salt.
Claim 21
Dependent Claim
15 words
The mixture as defined in claim 15 wherein said cationic dye is a phenanthridium dye.
Claim 22
Dependent Claim
14 words
The mixture as defined in claim 17 wherein said phenanthridium dye is ethidium bromide.
Claim 23
Dependent Claim
18 words
The mixture as defined in claim 15 wherein said sulfonated fluorescent derivative of 1,8 naphthalimide is brilliant sulfaflavine.
Claim 24
Dependent Claim
32 words
A dye mixture for differentially staining biological cells said mixture comprising: (a) a sulfonated triazinyl derivative of diamino stilbene; (b) a cationic dye; and (c) a toluidino derivative of naphthalene sulfonic acid.
Claim 25
Dependent Claim
32 words
A dye mixture for differentially staining biological cells said mixture comprising: (a) a sulfonated triazinyl derivative of diamino stilbene; (b) a cationic dye; and (c) a toluidino derivative of naphthalene sulfonic acid.
Claim 26
Dependent Claim
31 words
The mixture as defined in claim 20 wherein said sulfonated triazinyl derivative of diamino stilbene is 4,4' bis [4-(3 sulfoanilino)-6-[bis (2-hydroxy-ethyl) amino] 1,3,5 triazin-2yl] amino stilbene 2,2' disulfonic acid tetrasodium salt.
Claim 27
Dependent Claim
15 words
The mixture as defined in claim 20 wherein said cationic dye is a phenanthridium dye.
Claim 28
Dependent Claim
14 words
The mixture as defined in claim 22 wherein said phenanthridium dye is ethidium bromide.
Claim 29
Dependent Claim
19 words
The mixture as defined in claim 20 wherein said toluidino derivative of naphthelene sulfonic acid is 8-p-toluidino-1-naphthalene sulfonic acid.
Claim 30
Independent Claim
98 words
A method of staining a biological cell and comprising, differentially dyeing specific constituents of said cell with a mixture of a fluorescent dye capable of dyeing a first of said constituents, and a sensitizer compound capable of being coupled to a different one of said constituents and capable of absorbing incident radiation in a spectral absorption band and of transmitting to said dye bound to said first constituent adjacent to said sensitizer compound coupled to said different constituent, at least part of the energy of the absorbed incident radiation, thereby to excite said bound dye into fluorescent emission.
Claim 31
Independent Claim
98 words
A method of staining a biological cell and comprising, differentially dyeing specific constituents of said cell with a mixture of a fluorescent dye capable of dyeing a first of said constituents, and a sensitizer compound capable of being coupled to a different one of said constituents and capable of absorbing incident radiation in a spectral absorption band and of transmitting to said dye bound to said first constituent adjacent to said sensitizer compound coupled to said different constituent, at least part of the energy of the absorbed incident radiation, thereby to excite said bound dye into fluorescent emission.
References Cited (Backward Citations)
US3386920A
Cited
US3586859A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1982-03-26 | 1982-09-25 | 1983-03-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1986-03-26 | 1986-09-25 | 1987-03-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1990-03-26 | 1990-09-25 | 1991-03-27 | $7700.00 | Window Closed / Lapsed |