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Flow microfluorometric quantitation of the blastogenic response of lymphocytes

LS Cram, ER Gomez, CO Thoen, JC Forslund and JH Jett

A method for quantitating the specific stimulation of peripheral lymphocytes has been developed using the techniques of flow microfluorometry. Peripheral bovine lymphocytes were collected and specifically stained for deoxyribonucleic acid (DNA) content using a low-salt propidium iodide procedure. Flow microfluorometry was used to determine, on the basis of DNA content, the percentage of cells in a population that was stimulated. Extremely uniform staining of the lymphocytes (coefficient of variation of less than 2%) provides a high resolution between proliferating and nonproliferating cells. The method provides a rapid, highly repoducible technique for determing the fraction of lymphocytes stimulated in response to tuberculin antigens based on an increase in cellular DNA content. Specific and nonspecific stimulation by a defined antigen can be measured and resolved.

Volume 24, Issue 1, pp. 383-387, 01/01/1976
Copyright © 1976 by The Histochemical Society


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P. Horan and L. Wheeless Jr
Quantitative single cell analysis and sorting
Science, October 14, 1977; 198(4313): 149 - 157.
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