Flow cytometric quantification of all phases of the cell cycle and apoptosis in a two-color fluorescence plot.
An optimal technology for cell cycle analysis would allow the concomitant measurement of apoptosis, G0, G1, S, G2 and M phases in combination with cell surface phenotyping. We have developed an easy method in flow cytometry allowing this discrimination in an only two-color fluorescent plot. It is based on the concomitant use of 7-amino-actinomycin D and the antibodies anti-Ki67 and anti-phospho(Ser10)-histone H3, both conjugated to Alexa Fluor®488 to discriminate G0 and M phases, respectively. The method is particularly valuable in a clinical setting as verified in our laboratory by analyzing human leukemic cells from marrow samples or after exposure to cell cycle modifiers.
Université François-Rabelais de Tours, Tours, France.
Christine
C
Vignon
Christelle
C
Debeissat
Marie-Thérèse
MT
Georget
Didier
D
Bouscary
Emmanuel
E
Gyan
Philippe
P
Rosset
Olivier
O
Herault
D017209:Apoptosis / Q000187:drug effects; D001854:Bone Marrow Cells / Q000187:drug effects / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D003703:Demecolcine / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D016130:Immunophenotyping / Q000379:methods; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism; D050257:Tubulin Modulators / Q000494:pharmacology
D050257:Tubulin Modulators; D003703:Demecolcine