FULLY AUTOMATIC METHOD OF RAPIDLY IDENTIFYING AND CHARATERISING RETICULOCYTES, ERYTHROCYTES AND THROMBOCYTES IN WHOLE BLOOD AND COMPOSITION OF A REAGENT USED THEREFOR

摘要

The invention provides a very fast and reliable method and reagent composition for determining reticulocyte and erythrocyte counts, identification and characterization, as well as platelet counts, in a whole blood sample using fully automated hematology analyzers. The reagent composition includes a zwitterionic surfactant as sphering agent for reticulocytes and erythrocytes, an organic cationic dye, e.g., Oxazine 750, for staining the reticulocytes, and buffer solution components for maintaining a reagent pH of about 7.2 to 7.8, and an osmolality of about 292 +/- 5. On the basis of the particular pH, ionic strength, and dye concentration, the reagent composition of the present method improves upon previous methods, thus allowing fully automated blood sample analyses to be completed in less than about 30 seconds, with only about a 20 second incubation of sample in the reagent solution before analysis. With such a rapid incubation time, the reagent mixture is passed through the sensing region of a flow cytometer, and the light scattered and absorbed by each cell is rapidly measured to provide accurate cell determinations and counts. The volume and hemoglobin concentration and content of each erythrocyte or reticulocyte are determined. Mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes and/or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.