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Cell-Cycle Analysis of Asynchronous Populations

  • Michael G. Ormerod
Part of the Methods in Molecular Biology™ book series (MIMB, volume 263)

Abstract

Cells are incubated continuously in bromodeoxyuridine (BrdUrd), which is incorporated into cells synthesizing DNA. At intervals, cells are harvested and nuclei are prepared and stained with a bis-benzimidazole, Hoechst 33258, and propidium iodide. In the flow cytometer, the dyes are excited by UV and blue and red fluorescences recorded. BrdUrd quenches the blue fluorescence of the Hoechst dye. The degree of quenching records the progress of the cell through S phase(s); the red (PI) fluorescence yields the cell cycle phases. By this means, the progress of cells around the cell cycle can be followed and the effects of cytotoxic drugs, radiation, and other treatments observed.

Key Words

Bromodeoxyuridine flow cytometry Hoechst 33258 proliferation 

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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Michael G. Ormerod
    • 1
  1. 1.ReigateUK

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