Method and apparatus for classifying B-lymphocytes
First Claim
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1. A method for classifying B-lymphocytes, comprising the steps of:
- (1) preparing a measurement sample by mixing a blood sample with a lysing reagent to lyse erythrocytes and maintaining a mixture of said lysing reagent and said blood sample for from 15 to 60 minutes to shrink B-lymphocytes in said blood sample;
(2) flowing said measurement sample through a flow cell of a flow cytometer;
(3) radiating a light beam onto each individual cell in the measurement sample that is flowing through the flow cell;
(4) detecting at least two scattered light emitted from the irradiated cells, wherein said two scattered light are selected from the group consisting of forward scattered light, backward scattered light, and side scattered light;
(5) preparing a two-dimensional scattergram using the detected two scattered light by plotting intensities of the detected two scattered light from each of the cells;
(6) specifying a region of a B-lymphocyte cluster on the two-dimensional scattergram using the detected two scattered light; and
(7) counting the number of B-lymphocytes in the specified region.
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Abstract
A B-lymphocyte classifying method includes the steps of (1) preparing a measurement sample by mixing a blood sample with a lysing reagent to lyse erythrocytes and to shrink the B-lymphocytes in the blood sample, (2) letting the measurement sample flow through a flow cell of a flow cytometer, (3) radiating a light beam onto a cell in the measurement sample that is flowing through the flow cell, (4) detecting at least two scattered light emitted from the irradiated cell, (5) specifying a region of a B-lymphocyte cluster in accordance with the detected scattered light, and (6) counting the number of B-lymphocytes in the specified region.
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10 Claims
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1. A method for classifying B-lymphocytes, comprising the steps of:
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(1) preparing a measurement sample by mixing a blood sample with a lysing reagent to lyse erythrocytes and maintaining a mixture of said lysing reagent and said blood sample for from 15 to 60 minutes to shrink B-lymphocytes in said blood sample; (2) flowing said measurement sample through a flow cell of a flow cytometer; (3) radiating a light beam onto each individual cell in the measurement sample that is flowing through the flow cell; (4) detecting at least two scattered light emitted from the irradiated cells, wherein said two scattered light are selected from the group consisting of forward scattered light, backward scattered light, and side scattered light; (5) preparing a two-dimensional scattergram using the detected two scattered light by plotting intensities of the detected two scattered light from each of the cells; (6) specifying a region of a B-lymphocyte cluster on the two-dimensional scattergram using the detected two scattered light; and (7) counting the number of B-lymphocytes in the specified region. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A method for classifying B-lymphocytes, comprising the steps of:
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(1) preparing a measurement sample by mixing a blood sample with a lysing reagent to lyse erythrocytes and maintaining said lysing reagent and said blood sample for from 15 to 60 minutes to shrink B-lymphocytes in said blood sample; (2) flowing said measurement sample through a flow cell of a flow cytometer; (3) radiating a light beam onto each individual cell in the measurement sample that is flowing through the flow cell; (4) detecting at least two scattered light emitted from the irradiated cells, wherein said two scattered light are selected from the group consisting of forward scattered light, backward scattered light, and side scattered light; (5) specifying B-lymphocytes on the basis of intensities of the detected two scattered light from each of the cells; and (6) counting the number of the specified B-lymphocytes. - View Dependent Claims (10)
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Specification