Enrichment for submitotic cell populations using flow cytometry

Cytometry. 2000 Feb 1;39(2):126-30. doi: 10.1002/(sici)1097-0320(20000201)39:2<126::aid-cyto5>3.0.co;2-v.

Abstract

Background: One of the most dramatic events during the course of the mammalian cell cycle is mitosis, when chromosomes condense and segregate, the nuclear envelope breaks down, and the cell divides into two daughter cells. Although cells undergoing mitosis are cytologically distinguishable from nonmitotic cells, few molecular markers are available to specifically identify mitotic cells, especially cells within different stages of mitosis.

Methods: We applied the flow cytometric method of Juan et al. (Cytometry 32:71-77, 1998) to obtain cells with various levels of the molecular markers cyclin B1 and phosphorylated histone H3; fluorescence microscopy was then used to identify sorted cells in different stages of mitosis.

Results: We observed the substantial enrichment of submitotic cell populations.

Conclusions: This method represents an effective approach to obtain an enriched population of submitotic cells without the use of drug treatments or prior synchronization.

MeSH terms

  • Biomarkers / analysis
  • Cell Line
  • Cyclin B / analysis
  • Cyclin B1
  • Female
  • Flow Cytometry / methods*
  • Histones / analysis
  • Humans
  • Male
  • Microscopy, Fluorescence
  • Mitosis / genetics*
  • Phosphorylation

Substances

  • Biomarkers
  • CCNB1 protein, human
  • Cyclin B
  • Cyclin B1
  • Histones