Open AccessEstimation of membrane potentials of individual lymphocytes by flow cytometry.
Author(s) -
Howard M. Shapiro,
Peter J. Natale,
Louis A. Kamentsky
Publication year - 1979
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.76.11.5728
Subject(s) - valinomycin , ficoll , flow cytometry , membrane potential , depolarization , concanavalin a , biophysics , membrane , nigericin , chemistry , gramicidin , turbidimetry , ionophore , biology , chromatography , biochemistry , microbiology and biotechnology , in vitro , peripheral blood mononuclear cell
The membrane potentials of individual cells can be estimated by flow cytometric quantitation of the cells' uptake of the fluorescent lipophilic cationic dye 3,3'-dihexyloxacarbocyanine iodide. Human lymphocytes separated from peripheral blood on Hypaque-Ficoll gradients are uniformly depolarized by gramicidin and hyperpolarized by valinomycin. Concanavalin A and phytohemagglutinin depolarize only a fraction of the lymphocytes. The flow cytometric technique allows precise detection of heterogeneous membrane potential responses to stimuli such as lectins; it could also provide a basis for sorting cells that respond differently to a given stimulus.