Reticulocyte analyzing method and apparatus utilizing light scatter techniques
AssigneeCoulter Corporation
InventorsCarlos M. Rodriguez, Stephen L. Ledis
Life of this patent
Estimating the term…
Abstract
A reticulocyte analyzing method and apparatus in which a biological sample stream of ghosted red cells is passed into and through a point focused beam of light, such as laser light. A light detector structure is positioned with respect to the axis of the light beam to provide a light output pulse indicative of the passage of each cell. Electrically conductive contacts within the fluid stream can provide additional electrical pulse outputs of each cell. A staining reagent can be utilized with a ghosting reagent to further differentiate the reticulocytes. The light and the light and electronic produced output pulses or signals can be combined to define and quantify reticulocytes as distinct from other known cell types.
Claims24
A method of determining a percent of reticulocytes to the total number of erythrocytes in at least a portion of a whole blood sample in a cytometric flow cell, comprising:
- a. combining a portion of a whole blood sample, including a plurality of cells, with a RNA precipitating first dye, said first dye being used to delineate reticulocytes for light scatter detection of said reticulocytes;
- b. combining said portion of a whole blood sample including a plurality of cells with a red blood cell ghosting reagent which removes hemoglobin from said red blood cells;
- c. passing an aliquot of said combined whole blood sample, RNA precipitating first dye and ghosting reagent through a cytometric flow cell, including passing said cells substantially one at a time through a sensing zone in said cytometric flow cell and concurrently passing at least a beam of light through said sensing zone to intersect said cells;
- d. detecting light scattered caused by each cell as each cell intersects said beam of light;
- and
- e. analyzing upper median angle light scatter (UMALS) of said light scattered by each cell to obtain a determination of the percentage of reticulocytes to the total number of erythrocytes in said whole blood sample.
An apparatus for determining a percent of reticulocytes to the total number of erythrocytes in at least a portion of a whole blood sample in a cytometric flow cell, comprising:
- a. means for combining a portion of a whole blood sample, including a plurality of cells, with a RNA precipitating first dye, said first dye being used to delineate reticulocytes for light scatter detection of said reticulocytes;
- b. means for combining said portion of a whole blood sample, including a plurality of cells, with a red blood cell ghosting reagent which removes hemoglobin from said red blood cells;
- c. means for passing an aliquot of said combined whole blood sample, RNA precipitating first dye and ghosting reagent through a cytometric flow cell, wherein said flow cell includes means for passing said cells substantially one at a time through a sensing zone in said cytometric flow cell and concurrently passing at least a beam of light through said sensing zone to intersect said cells;
- d. means for detecting light scattered caused by each cell as each cell intersects said beam of light;
- and
- e. means for analyzing upper median angle light scatter(UMALS) of said light scattered by each cell to obtain a determination of the percentage of reticulocytes to the total number of erythrocytes in said whole blood sample.
A method of determining reticulocyte count in at least a portion of a whole blood sample in a cytometric flow cell, comprising:
- a. combining a portion of a whole blood sample, including a plurality of cells, with a RNA precipitating first dye, said first dye being used to delineate reticulocytes for facilitating light scatter detection of said reticulocytes;
- b. combining said portion of a whole blood sample including a plurality of cells with a red blood cell ghosting reagent which removes hemoglobin from said red blood cells;
- c. passing an aliquot of said combined whole blood sample, RNA precipitating first dye and ghosting reagent through a cytometric flow cell, including passing said cells substantially one at a time through a sensing zone in said cytometric flow cell and concurrently passing at least a beam of light through said sensing zone to intersect said cells;
- d. detecting light scattered caused by each cell as each cell intersects said beam of light;
- and
- e. analyzing upper median angle light scatter (UMALS) of said light scattered by each cell to obtain a determination of the number of reticulocytes in said blood sample.
An apparatus for determining reticulocyte count in at least a portion of a whole blood sample in a cytometric flow cell, comprising:
- a. means for combining a portion of a whole blood-sample,. including a plurality of cells, with a RNA precipitating first dye, said first dye being used to delineate reticulocytes for light scatter detection of said reticulocytes;
- b. means for combining a portion of a whole blood sample, including a plurality of cells with a red blood cell ghosting reagent which removes hemoglobin from said red blood cells;
- c. means for passing an aliquot of said combined whole blood sample, RNA precipitating first dye and ghosting reagent through a cytometric flow cell, wherein said flow cell includes means for passing said cells substantially one at a time through a sensing zone in said cytometric flow cell and concurrently passing at least a beam of light through said sensing zone to intersect said cells;
- d. means for detecting light scattered caused by each cell as each cell intersects said beam of light;
- and
- e. means for analyzing upper median angle light scatter (UMALS) of said light scattered by each cell to obtain a determination of the number of reticulocytes in said blood sample.
Dependencies are inferred from the claim text.