Estimating cell death in G2M using bivariate BrdUrd/DNA flow cytometry

R. Allen White, David Asmuth, Ying Lu, Nan Wang, Xiao Dong Li, Lisa Reece, Richard B Pollard, Mostafa Nokta, James F. Leary, Nicholas H A Terry

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Background: In an accompanying paper (Asmuth et al.) it was found necessary to include cell death explicitly to estimate parameters of cell proliferation. The use of bivariate flow cytometry to estimate the phase durations and the doubling times of cells labeled with thymidine analogues is well established. However, these methods of analysis do not consider the possibility of cell death. This report demonstrates that estimating cell death in G2/M is possible. Methods: Mathematical models for the experimental quantities, the fraction of labeled undivided cells, the fraction of labeled divided cells, and the relative movement were developed. These models include the possibility that, of the cells with G2/M DNA content, only a certain fraction will divide, with the remainder dying after some time TR. Simulation studies were conducted to test the possibility of using simple methods to estimate phase durations and cell death rates. Results: Cell death alters the estimates of phase transit times in a rather complex manner that depends on the lifetime of the doomed cells. However, it is still possible to obtain estimates of the phase durations of cells in S and G2/M and the death rates of cells in G2/M. Conclusions: The methods presented herein provide a new way to characterize cell populations that includes cell death rates and common measurements of cell proliferation.

Original languageEnglish (US)
Pages (from-to)32-40
Number of pages9
JournalCytometry Part A
Volume66
Issue number1
DOIs
StatePublished - Jul 2005

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Keywords

  • Bromodeoxyuridine
  • Cell cycle
  • Phase durations
  • Population growth

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

White, R. A., Asmuth, D., Lu, Y., Wang, N., Li, X. D., Reece, L., Pollard, R. B., Nokta, M., Leary, J. F., & Terry, N. H. A. (2005). Estimating cell death in G2M using bivariate BrdUrd/DNA flow cytometry. Cytometry Part A, 66(1), 32-40. https://doi.org/10.1002/cyto.a.20147