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US 5,125,737 AGranted
Multi-part differential analyzing apparatus utilizing light scatter techniques
AssigneeCoulter Electronics, Inc.
InventorsCarlos M. Rodriguez, Wallace H. Coulter
Life of this patent
Estimating the term…
Abstract
Disclosed is a flow through particle analyzing cell differentiating apparatus for optical and electronic measurements on a stream of particles in which a hydrodynamically focoussed stream is passed into and through a point focussed beam of radiated energy whereby the beam is scattered by the stream to impinge upon light responsive signal operating members disposed adjacent the cell at angular positions relative to the beam axis of from about 10.degree. to 70.degree..
Claims40
Apparatus for selectively differentiating at least one white blood cell subpopulation in a blood sample, whereby differentiation is based upon individual cell light scatter, said apparatus comprising:
- a cytometric flow cell including inlet and outlet means for the ingress and egress of a blood sample in liquid suspension;
- introducing means for introducing a blood sample including white cells into said flow cell inlet to cause said white cells to flow through said flow cell;
- a beam of light arranged such that the axis of its light rays pass through said flow cell at right angles to said flow of white cells;
- extinguishing means, axially aligned with said axis of light rays, for extinguishing the light rays which are at a low angle to said beam axis which are scattered by the white cells in response to the impingement of light thereon as they pass through said beam;
- light collection means responsive to the light scattered by said cells in a median angle (MALS) greater than 10.degree., but less than 70.degree., relative to said axis after passing through said beam for producing median angle output signal data, eosinophil subpopulation data being generated by and derived from the scattered rays in the (MALS) range of 20.degree. to 70.degree.;
- utilizing means for utilizing said output signal data from said light collection means for utilizing the portion of said (MALS) in the range of 20.degree. to 70.degree. to differentiate the eosinophil subpopulation;
- and
- said apparatus is constructed to operate to differentiate white blood cells, especially including eosinophil blood cells, as a result of the inherent, natural characteristics of the white blood cells and without treatment of any kind for specifically altering the inherent, natural characteristics of the white blood cells for differentiating eosinophils.
5 more elements
A method for differentiating at least one white blood cell subpopulation in a blood sample, whereby differentiation is based upon individual cell light scatter, said method comprising the steps of:
- passing a beam of light along an axis and orthogonally through a fluid passageway while simultaneously flowing said blood sample through said passageway;
- extinguishing the rays of light which are at a low angel to said axis which are scattered by the white cells in response to impingement of light thereon as they pass through said beam of light;
- collecting the light scattered by said cells in a median angle (MALS) greater than 10.degree., but less than 70.degree., relative to said axis after passing through said beam of light;
- sensing the rays of light scattered in said median angle by each cell of said blood sample for producing median angle output signal data;
- said step of sensing providing signal data which is a result of the inherent, natural light scatter characteristics of the white blood cells without treatment of any kind for specifically altering the inherent, natural characteristics of said white blood cells for differentiating eosinophils, eosinophil data being contributed by the portion of (MALS) from 20.degree. to 70.degree.;
- and utilizing said portion of said median angle light scatter signal data to differentiate the eosinophil subpopulation.
3 more elements
A method for differentiating at least one white blood cell subpopulation in a blood sample, whereby differentiation is based upon individual cell light scatter, said method comprising the steps of:
- passing rays of light along an axis and orthogonally through a fluid passageway while simultaneously flowing said blood sample through said passageway;
- extinguishing the rays of light which are at a low angle to said axis which are scattered by the white cells in response to impingement of light therein as they pass through said beam of light;
- collecting the light scattered by said cells in a median angle (MALS) greater than 10.degree., but less than 70.degree., relative to said axis after passing through said beam of light;
- sensing the rays of light scattered in said median angle for producing median angle output signal data by each cell of said blood sample;
- eosinophil subpopulation data being generated by and derived from the scattered rays in the (MALS) range of 20.degree. to 70.degree.;
- said step of sensing providing signal data as a result of the inherent, natural light scatter characteristics of the white blood cells without treatment of any kind for specifically altering the inherent, natural characteristics of said white blood cells for differentiating white blood cell subpopulations;
- and processing said output signal data to produce data for utilizing the portion of said (MALS) in the range of 20.degree. to 70.degree. to differentiate at least three subpopulations, namely eosinophils, neutrophils, and lymphocytes, monocytes and basophils.
4 more elements
A method for differentiating at least one white blood cell subpopulation in a blood sample, whereby differentiation is based upon individual cell light scatter, said method comprising the steps of:
- passing rays of light along an axis and orthogonally through a fluid passageway while simultaneously flowing said blood sample through said passageway;
- extinguishing the rays of light which are at a low angle to said axis which are scattered by the white cells in response to impingement of light thereon as they pass through said beam of light;
- collecting the light scattered by said cells in a median angle (MALS) greater than 10.degree., but less than 70.degree., relative to said axis after passing through said beam of light;
- sensing the rays of light scattered in said median angle by each cell of said blood sample for producing median angle output signal data;
- said step of sensing provides signal data as a result of the inherent, natural light scatter characteristics of the white blood cells without treatment of any kind for specifically altering the inherent, natural characteristics of said white blood cells for differentiating white blood cell subpopulations and is within a collection angle of from 10.degree. to 70.degree. relative to said axis;
- processing electronically said output signal data to produce white blood cell differentiating data;
- wherein said step of sensing is operative with the untreated white blood cells and within a collection angle of from 10.degree. to 70.degree. relative to said axis;
- eosinophil subpopulation data being generated by and derived from the scattered rays in the (MALS) range of 20.degree. to 70.degree.;
- and including the additional steps of obtaining axial light loss signal information (ALL), generating Coulter volume DC signal information (DC), and comparing said ALL with said DC for producing data for differentiating three white blood cell subpopulations, namely:
- lymphocytes, basophils and eosinophils, monocytes and neutrophils.
7 more elements
A method for differentiating at least one white blood cell subpopulation in a blood sample, whereby differentiation is based upon individual cell light scatter, said method comprising the steps of:
- passing rays of light along an axis and orthogonally through a fluid passageway while simultaneously flowing said blood sample through said passageway;
- extinguishing the rays of light which are at a low angle to said axis which are scattered by the white cells in response to impingement of light thereon as they pass through said beam of light;
- collecting the light scattered by said cells in a median angle (MALS) greater than 10.degree., but less than 70.degree., relative to said axis after passing through said beam of light;
- sensing the rays of light scattered in said median angle by each cell of said blood sample for producing median angle output signal data;
- said step of sensing providing signal data as a result of the inherent, natural light scatter characteristics of the white blood cells without treatment of any kind for specifically altering the inherent, natural characteristics of said white blood cells for differentiating white blood cell subpopulations;
- processing electronically the said output signal data to produce white blood cell differentiating data;
- wherein said step of sensing is operative with the untreated white blood cells and within a collection angle of from 10.degree. to 70.degree. relative to said axis;
- and including the steps of applying a DC and an RF current simultaneously to said flow cell passageway aperture producing a Coulter volume DC signal (DC) and a Coulter RF signal (RF), and comparing said DC to said RF, whereby said processing provides data for differentiating at least four white blood cell subpopulations, namely:
- neutrophils, monocytes, lymphocytes, and basophils.
6 more elements
A method for differentiating at least one white blood cell subpopulation in a blood sample, whereby differentiation is based upon individual cell light scatter, said method comprising the steps of:
- passing rays of light along an axis and orthogonally through a fluid passageway including a Coulter aperture while simultaneously flowing said blood sample through said passageway;
- sensing some of the rays of light scattered by each cell of said blood sample;
- processing electronically the results of said sensing step to produce white blood cell differentiating data;
- wherein said step of sensing provides median angle light scatter (MALS) signal data throughout a range of approximately 10.degree.-70.degree. and wherein said processing step develops a log of median angle light scatter signal (log of MALS);
- including the steps of applying a DC current simultaneously to said flow cell passageway aperture producing a Coulter volume DC signal (DC), dividing the log of MALS by said DC for generating rotated light scatter signal information (RLS) and comparing said DC with said RLS, whereby said processing provides data for differentiating and identifying at least four white blood cell subpopulations, namely:
- lymphocytes and basophils, monocytes, neutrophils, and eosinophils.
3 more elements
A method for differentiating at least one white blood cell subpopulation in a blood sample, whereby differentiation is based upon individual cell light scatter, said method comprising the steps of:
- passing rays of light along an axis and orthogonally through a fluid passageway including a Coulter aperture while simultaneously flowing said blood sample through said passageway, sensing some of the rays of light scattered by each cell of said blood sample;
- processing electronically the results of said sensing step to produce white blood cell differentiating data, wherein said step of sensing provided median angle light scatter (MALS) signal data throughout a range of approximately 10.degree.-70.degree. and said processing step develops a log of median angle light scatter signal (log of MALS), applying a DC and an RF current simultaneously to said flow cell passageway aperture for producing a Coulter volume DC signal (DC), dividing the log of MALS by said DC for generating rotated light scatter signal information (RLS), and comparing said DC with said RLS, whereby said processing provides data for differentiating and identifying at least four white blood cell subpopulations, namely:
- lymphocytes and basophils, monocytes, neutrophils, and eosinophils, dividing said RF by said DC for generating opacity signal information (OP), and comparing said DC with said OP while gating on said RLS, whereby said processing provides data for further differentiating and identifying one of said four white blood cell subpopulations into at least two white blood cell subpopulations, namely:
- lymphocytes and basophils.
1 more element
A method for differentiating at least one white blood cell subpopulation in a blood sample, whereby differentiation is based upon individual cell light scatter, said method comprising the steps of:
- passing rays of light along an axis and orthogonally through a fluid passageway including a Coulter aperture while simultaneously flowing said blood sample through said passageway;
- sensing some of the rays of light scattered by each cell of said blood sample;
- processing electronically the results of said sensing step to produce white blood cell differentiating data;
- wherein said step of sensing provides median angle light scatter (MALS) signal data throughout a range of approximately 10.degree.-70.degree. and said processing step develops a log of median angle light scatter signal (log of MALS);
- applying a DC and an RF current simultaneously to said flow cell passageway aperture for producing a Coulter volume DC signal (DC), dividing the log of MALS by said DC for generating rotated light scatter signal information (RLS), dividing said RF by said DC for generating opacity signal information (OP), and comparing said DC with said RLS and said OP, whereby said processing provides data for differentiating and identifying at least five white blood cell populations, namely:
- lymphocytes, basophils, monocytes, neutrophils, and eosinophils.
3 more elements
Dependencies are inferred from the claim text.