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 ba...

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Auteurs principaux: Christine Vignon, Christelle Debeissat, Marie-Thérèse Georget, Didier Bouscary, Emmanuel Gyan, Philippe Rosset, Olivier Herault
Format: article
Langue:EN
Publié: Public Library of Science (PLoS) 2013
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Accès en ligne:https://doaj.org/article/15449e40bade46b0b85c696e0fc59e71
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Résumé: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.