A flow cytometric technique for simultaneous analysis of human mononuclear cell surface antigens and DNA.
A method for dual staining of mononuclear cells for lymphocyte phenotypic markers and DNA is described. The cells were stained with fluorescein-conjugated monoclonal antibodies and then rendered permeable to propidium iodide using saponin. Propidium iodide stains DNA and, using flow cytometry, cell cycle analysis of individual lymphocyte subpopulations can be determined. Saponin acts within 1 min, preserves expression of surface antigens and is effective at all concentrations from 0.001% to 1%. This technique is simple, rapid and gives reproducible results.
Rigg
K M
KM
Department of Surgery, Medical School, Newcastle upon Tyne, U.K.
Shenton
B K
BK
Murray
I A
IA
Givan
A L
AL
Taylor
R M
RM
Lennard
T W
TW
D000954:Antigens, Surface / Q000032:analysis*; D002453:Cell Cycle; D002462:Cell Membrane / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D004247:DNA / Q000032:analysis*; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000032:analysis*; D012503:Saponins / Q000494:pharmacology*
D000954:Antigens, Surface; D012503:Saponins; D004247:DNA