Patent ID: 6524858
Filing Date: 2003-02-25
Classification: G01N,Y10T

Abstract:
A single channel, single dilution method for identifying and measuring cell components in a normal or abnormal mammalian blood sample, comprising mature red blood cells; reticulocytes; white blood cells, including lymphocytes, basophils, eosinophils, monocytes, and neutrophils; platelets and reticulated platelets, and for determining qualitative and quantitative parameters of said blood sample components, comprising:(a) mixing in a single dilution an aliquot of the blood sample with an aqueous reagent composition to form a reaction mixture, said reagent composition comprising a surfactant as sphering agent in an amount effective to sphere blood cells in the sample; a dye compound in an amount effective to stain reticulocyte RNA and white blood cell nucleic acid; and a buffer or buffer solution; wherein the red blood cells remain substantially unlysed in the reaction mixture; (b) passing the reaction mixture of (a) substantially one cell at a time through a flow cell in a single channel, wherein light is scattered and absorbed by each cell component; said scattered light being optically detected at a low angle interval of about 1 to 10 degrees to produce a low light scatter intensity measurement and at a high angle interval of about 4 to 30 degrees to produce a high light scatter intensity measurement; (c) detecting absorption signals produced by incident radiation resulting from binding of the dye compound to reticulocyte and white blood cell nucleic acid, or components thereof, in the single channel; and (d) discriminating among and measuring each of the different blood cell components, comprising mature red blood cells; reticulocytes; white blood cells, including lymphocytes, basophils, eosinophils, monocyte, and neutrophils; platelets and reticulated platelets of the sample by detecting scatter-scatter optical signals, scatter-absorption optical signals, or scatter-scatter-absorption optical signals by means of the scattering measurements and absorption signals of steps (b) and (c); wherein detection and measurement are performed on each of the cell components flowing through the flow cell in the single channel.