Rapid identification of T helper and T cytotoxic/suppressor lymphocytes with an oxazine dye.
Peripheral blood lymphocytes displayed a plurality of sizes and colors when exposed first to a methanolic solution of C.I. basic blue 141, then to an aqueous alkaline solution of the same dye and rinsed in a neutral HEPES buffer containing trace amounts of various salts. As confirmed with purified lymphocyte subpopulations obtained with a cell sorter, T helper cells (CD4) were small and their nuclei and cytoplasm stained deep blue. T cytotoxic/suppressor cells (CD 8) were larger than T helper cells, their nuclei stained pale green or blue green and their cytoplasm contained a cluster of magenta colored granules. From start to finish, the stain takes 15 min to perform. Used in the manner described, basic blue 141 holds promise as a rapid means of identifying and differentiating CD4 and CD8 cells under the ordinary light microscope without using monoclonal antibodies or fluorescence.
Kass
L
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Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44109.
D006801:Humans; D016131:Lymphocyte Subsets; D010078:Oxazines; D013194:Staining and Labeling / Q000379:methods; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology*; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology*; D050378:T-Lymphocytes, Regulatory / Q000166:cytology*
D010078:Oxazines; C053870:C.I. Basic Blue 141