Flow cytometric quantification of all phases of the cell cycle and apoptosis in a two-color fluorescence plot.

Vignon, Christine; Debeissat, Christelle; Georget, Marie-Thérèse; Bouscary, Didier; Gyan, Emmanuel; Rosset, Philippe; Herault, Olivier · PLoS One · 2013

basic_science · Level V

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Abstract

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.

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