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. 2015 Jul 1:111:28.6.1-28.6.11.
doi: 10.1002/0471142727.mb2806s111.

Assaying Cell Cycle Status Using Flow Cytometry

Kang Ho Kim  1 , Joel M Sederstrom  2
Affiliations

Assaying Cell Cycle Status Using Flow Cytometry

Kang Ho Kim et al. Curr Protoc Mol Biol. .

Abstract

In this unit, two protocols are described for analyzing cell cycle status using flow cytometry. The first is based on the simultaneous analysis of proliferation-specific marker (Ki-67) and cellular DNA content, which discriminate resting/quiescent cell populations (G0 cell) and quantify cell cycle distribution (G1, S, or G2/M), respectively. The second is based on differential staining of DNA and RNA through co-staining of Hoechst 33342 and Pyronin Y, which is also useful to identify G0 cells from G1 cells. Along with these methods for analyzing cell cycle status, two additional methods for cell proliferation assays with recent updates of newly developed fluorophores, which allow multiplex analysis of cell cycle status, cell proliferation, and a gene of interest using flow cytometry, are outlined.

Keywords: Hoechst 33342; Ki-67; Pyronin Y; cell cycle; flow cytometry; propidium iodide.

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Figures

Figure 1
Figure 1
Analysis of cell cycle status by differential staining of Ki-67/DNA and DNA/RNA. HeLa cells were fixed and subsequently stained with Ki-67-FITC and PI (A) or Hoechst 33342 and Pyronin Y (B) according to the BASIC PROTOCOL 1 and 2.
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