METHOD FOR CLASSIFYING WHITE BLOOD CELL BY FLOW CYTOMETRY

摘要

PURPOSE:To discriminate and count the kinds of respective white blood cells by passing a colored sample to a flow cytometer, discriminating the white blood cells and the other blood cells and ghosts by fluorescent light intensity and measuring the fluorescent signal and side scattered light signal of the white blood cells. CONSTITUTION:Fresh blood subjected to an anticlotting treatment is added to a hypotonic 1st liquid consisting of dye neutral red which specifically strongly dyes eosinophil, dye anstrazone orange G which specifically strongly dyes basophil and buffer agent for maintaining pH in an acidic region to hemolyze the red blood cells. A 2nd liquid consisting of a buffer for neutralizing the acid in the buffer and maintaining the soln. pH at dye pH and osmotic pressure compensating agent for maintaining the soln. in the form of the white blood cells is added to the 1st liquid to dye the white blood cells. Such liquid is passed through the flowcyte meter to discriminate the white blood cells and the other blood cells and ghosts by the fluorescent intensity. The fluorescent signal and side (90 deg.) scattered light signal are then measured. The kinds of the respective white blood cells are identified by these signals.