EP0193356A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Method for analysis of subpopulations of blood cells
Assignee: Becton Dickinson and Co
•
Filing: 1986-02-20
•
Granted: —
•
Expires: 2006-02-20
Abstract
Abstract A method for identifying and enumerating cells of a subclass of blood cells in relation to cells of another subclass of blood cells is provided. In the method a first subclass of blood cells is selectively tagged by incubating an aliquot of a blood sample with a first tagging agent. A second subclass of blood cells is selectively tagged by incubating the aliquot with a second tagging agent. The aliquot is then passed, without lysing of any subclasses of blood cells which are not of interest, substantially a cell at a time through an area of optical stimulation for the tagging agents. Light emitted by the tagging agents is detected, the detection being limited by gating to a threshold value related to a predetermined intensity of light by one of the tagging agents. Cells of the subclass are differentiated based on occurrence of emitted light from the tagging agents.
Inventors
- 👤 Diether J. Recktenwald
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 21:09
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; G01N33/56966; G01N33/56972; G01N15/00; G01N15/10; G01N15/14; G01N15/1456; G01N15/1459; G01N33/5005; G01N33/5094; G01N15/01; G01N2015/1402; G01N2015/1477; G01N21/00; G01N21/62; G01N21/63; G01N21/64; G01N21/6428; Y; Y10; Y10S; Y10S435/00; Y10S435/968; Y10S436/00; Y10S436/80; Y10S436/827
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
236 words
A method for identifying and enumerating cells of a subclass of blood cells in relation to cells of another subclass of blood cells comprising the steps of: (1) providing an aliquot from a blood sample, (2) selectively tagging cells of a first subclass of blood cells by incubating said aliquot with a first tagging agent which is selectively reactive with cells of said first subclass, said first tagging agent having a measurable physical response to suitable stimulation; (3) selectively tagging cells of at least a second subclass of blood cells by incubating said aliquot with a second tagging agent which is selectively reactive with cells of said second subclass, said second tagging agent having a measurable physical response to suitable stimulation; (4) analyzing said aliquot, substantially a cell at a time, by subjecting said aliquot to suitable stimulation for both said first tagging agent and said second tagging agent while detecting said measurable physical response provided by said first tagging agent and said second tagging agent and capturing the data provided by said detection, said captured data being restricted to a predetermined range establishing the intensity of the physical response conferred on the cells of interest by at least one of said tagging agents, and (5) differentiating cells of said first subclass of cells from said second subclass based on occurrence of said measurable physical response of said first tagging agent and said second tagging agent.
Claim 2
Independent Claim
236 words
A method for identifying and enumerating cells of a subclass of blood cells in relation to cells of another subclass of blood cells comprising the steps of: (1) providing an aliquot from a blood sample, (2) selectively tagging cells of a first subclass of blood cells by incubating said aliquot with a first tagging agent which is selectively reactive with cells of said first subclass, said first tagging agent having a measurable physical response to suitable stimulation; (3) selectively tagging cells of at least a second subclass of blood cells by incubating said aliquot with a second tagging agent which is selectively reactive with cells of said second subclass, said second tagging agent having a measurable physical response to suitable stimulation; (4) analyzing said aliquot, substantially a cell at a time, by subjecting said aliquot to suitable stimulation for both said first tagging agent and said second tagging agent while detecting said measurable physical response provided by said first tagging agent and said second tagging agent and capturing the data provided by said detection, said captured data being restricted to a predetermined range establishing the intensity of the physical response conferred on the cells of interest by at least one of said tagging agents, and (5) differentiating cells of said first subclass of cells from said second subclass based on occurrence of said measurable physical response of said first tagging agent and said second tagging agent.
Claim 3
Dependent Claim
25 words
A method in accordance with Claim 1 wherein said first tagging agent and said second tagging agent have a predetermined fluorescent response to optical stimulation.
Claim 4
Dependent Claim
36 words
A method in accordance with Claim 2 wherein said aliquot is analyzed by passing said aliquot through an area of optical stimulation while detecting light emitted from said first tagging agent and said second tagging agent.
Claim 5
Dependent Claim
35 words
A method in accordance with Claim 1 wherein said first subclass of blood cells is _a set of white blood cells and said second subclass is a group of white blood cells within said set.
Claim 6
Dependent Claim
30 words
A method in accordance with Claim 1 wherein said first and second tagging agents are selected from the group consisting of antibodies, lectins, nucleic acid dyes and membrane potential dyes.
Claim 7
Dependent Claim
17 words
A method in accordance with Claim 1 wherein said first and second tagging agents are monoclonal antibodies.
Claim 8
Dependent Claim
38 words
A method in accordance with Claim 1 wherein at least one of the group consisting of said first tagging agent and said second tagging agent is reactive with distinct antigenic determinants of a subclass of cells of interest.
Claim 9
Dependent Claim
89 words
A method in accordance with Claim 1 or Claim 6 wherein said first subclass is selected from the group consisting of nucleated cells, white blood cells, lymphocytes, granulocytes, T-cells, B-cells and NK/ LGL cells and said second subclass of blood cells is selected from the group consisting of lymphocytes, granulocytes, monocytes, helper T-cells, inducer T-cells, suppressor T-cells, cytotoxic T -cells, T-cells, B-cells, T-cells in maturation/ activation stages, B-cells in maturation/activation stages, monocytes in maturation/activation stages, neutrophils, basophils, eosinophils and granulocytes -in maturation/activation stages and NK/LGL cells in maturation/activation stages.
Claim 10
Dependent Claim
53 words
A method in accordance with Claim 1 or Claim 2 wherein at least a third subclass of blood cells is selectively tagged by incubating said aliquot with a third tagging agent which is selectively reactive with cells of said third subclass, said third tagging agent having a measurable physical response to suitable stimulation.
Claim 11
Dependent Claim
20 words
A method in accordance with Claim 9 wherein said third tagging agent has a predetermined fluorescent response to optical stimulation.
Claim 12
Dependent Claim
16 words
A method in accordance with Claim 10 wherein said third tagging agent is a monoclonal antibody.
Claim 13
Dependent Claim
18 words
A method in accordance with Claim 11 wherein one of said tagging agents is a nucleic acid dye.
Claim 14
Dependent Claim
26 words
A method in accordance with Claim 1 or Claim 10 wherein said tagging agents are selected from antibodies, lectins, nucleic acid dyes and membrane potential dyes.
Claim 15
Dependent Claim
35 words
A method in accordance with Claim 2 or Claim 10 wherein said tagging agents are marked with a marking agent selected from the group consisting of phycoerythrin, fluorescein isothiocyanate, texas red, allophyocyanine, R-phycocyanin and LDS-741.
Claim 16
Dependent Claim
33 words
A method in accordance with Claim 3 wherein said aliquot is passed through said area of opticaL stimulation at a rate of from about 20 to about 2000 white blood cells per second.
Claim 17
Dependent Claim
37 words
A method in accordance with Claim 1 wherein said first and second subclasses are white blood cells and subclasses thereof and said first and second tagging agents are monoclonal antibodies - . conjugated to fluorescent marking agents.-
Claim 18
Dependent Claim
47 words
A method in accordance with Claim 9 wherein said first, second and third subclasses are white blood cells and subclasses thereof and at least one of the group consisting of said first tagging agent, said second tagging agent and said third tagging agent is a monoclonal antibody.
Claim 19
Dependent Claim
17 words
A method in accordance with Claim 1 or Claim 9 wherein said incubation steps are performed sequentially.
Claim 20
Dependent Claim
17 words
A method in accordance with Claim 1 or Claim 9 wherein said incubation steps are performed simultaneously.
Claim 21
Dependent Claim
25 words
A method in accordance with Claim 1 or Claim 9 wherein said aliquot is analyzed without lysis or separation of any subclass of blood cells.
Claim 22
Dependent Claim
39 words
A method in accordance with Claim 1 or Claim 9 wherein at least one of said measurable physical responses of said tagging agents is provided by a physical property associated with at least one of said subclasses of cells.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.