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

PLoS One. 2013 Jul 30;8(7):e68425. doi: 10.1371/journal.pone.0068425. Print 2013.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis* / drug effects
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / metabolism
  • Cell Cycle* / drug effects
  • Cell Line
  • Demecolcine / pharmacology
  • Flow Cytometry* / methods
  • Humans
  • Immunophenotyping / methods
  • Lymphocytes / drug effects
  • Lymphocytes / metabolism
  • Tubulin Modulators / pharmacology

Substances

  • Tubulin Modulators
  • Demecolcine

Grants and funding

This research was supported by the the Ligue Nationale Contre le Cancer and the International Rotary Club of Blois. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.