Open AccessSimultaneous measurement of neutrophil, lymphocyte, and monocyte glutathione by flow cytometry
Author(s) -
Scott Robert B.,
Collin James M.,
Matin Sina,
White Frances,
Swerdlow Paul S.
Publication year - 1990
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1860040503
Subject(s) - glutathione , flow cytometry , monocyte , glutathione reductase , lymphocyte , chemistry , fluorescence , thiol , microbiology and biotechnology , biochemistry , biology , immunology , glutathione peroxidase , enzyme , physics , quantum mechanics
A flow cytometric method for quantitation of glutathione (GSH) was applied to simultaneous analysis of the major leukocyte types in peripheral blood. Cellular thiols (predominately GSH) were stained with monochlorobimane (MCIB), and thiol fluorescence was measured with a flow cytometer. The fluorescence of the thiols closely reflected the GSH content, as measured by a specific glutathione reductase assay. Fluorescence of individual cell types could be measured after delineating those cells by their light‐scatter characteristics, utilizing dual‐angle light scatter for discrimination. By this means, GSH contents of 12.5 ± 2.0nmol/10 7 neutrophils, 14.5 ± 2.7 nmol/10 7 monocytes, and 5.0 ± 1.0 nmol/10 7 lymphocytes were found. The results obtained for neutrophils with the flow cytometer were virtually identical with those obtained with chemical assay in purified samples of neutrophils, indicating the validity of the flow cytometric method.