Open AccessFlow microfluorometric quantitation of the blastogenic response of lymphocytes.
Author(s) -
L. Scott Cram,
Enrique R Santiago Gómez,
Charles O. Thoen,
J Forslund,
James H. Jett
Publication year - 1976
Publication title -
journal of histochemistry and cytochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.971
H-Index - 124
eISSN - 1551-5044
pISSN - 0022-1554
DOI - 10.1177/24.1.768372
Subject(s) - propidium iodide , chemistry , staining , microbiology and biotechnology , flow cytometry , dna , stimulation , population , antigen , dna synthesis , lymphocyte , chromatography , immunology , pathology , biology , biochemistry , endocrinology , medicine , apoptosis , environmental health , programmed cell death
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.
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