Single channel, single dilution detection method for the identification and quantification of blood cells and platelets in a whole blood sample using an automated hematology analyzer

Abstract

The invention provides a new single channel, single dilution method and system for identifying, analyzing and quantifying the cellular components of whole blood using a single channel, rather than multiple channels, of an automated hematology analyzer utilizing flow cytometry and the detection of the light scattered and absorbed by each cell. The single channel utilized in the method was previously known and used only for red blood cell and reticulocyte analysis. The method involves the use of an organic dye in the reagent solution for staining the nucleic acid of reticulocytes, including reticulated platelets, and white blood cells in the sample. The single channel method developed and described is particularly useful for determining white blood cell counts and assessing parameters of a whole blood sample, for blood samples from both human and non-human mammals. The single channel method of the invention employs only one blood diluent reagent composition that is mixed with the blood sample aliquot for analysis and, optimally, a sheath/rinse reagent, thus making it simpler and easier to use. In addition, the reagent compositions of the method are more benign to the white blood cells than previous reagents, which contained various lytic surfactants for removing red blood cells, for example. The apparatus for performing the method provides an economical, streamlined and space-saving analyzer for carrying out complete blood cell analysis on a mammalian whole blood sample.