Unit

UNIT 5.7 Analysis of Cellular DNA Content by Flow Cytometry

  1. Zbigniew Darzynkiewicz,
  2. Xuan Huang

Published Online: 1 MAY 2004

DOI: 10.1002/0471142735.im0507s60

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Darzynkiewicz, Z. and Huang, X. 2004. Analysis of Cellular DNA Content by Flow Cytometry. Current Protocols in Immunology. 60:5.7:5.7.1–5.7.18.

Author Information

  1. New York Medical Center, Valhalla, New York

Publication History

  1. Published Online: 1 MAY 2004
  2. Published Print: APR 2004

Abstract

Cellular DNA content can be measured by flow or laser-scanning cytometry with the aim of (1) revealing cell distribution within the major phases of the cell cycle, (2) estimating the frequency of apoptotic cells with fractional DNA content, and/or (3) disclosing the DNA-ploidy of the measured cell population. In this unit, simple and universally applicable methods for staining fixed cells are presented, as are methods that utilize detergents and proteolytic treatment to permeabilize cells. Additionally, supravital cell staining with Hoechst 33342, which is primarily used for sorting live cells for subsequent culturing based on DNA-content differences, is also described. Also presented are methods for staining of cell nuclei isolated from paraffin-embedded tissues, and deconvolution of DNA-content-frequency histograms to estimate the percentage of cells in major phases of the cell cycle and frequency of apoptotic cells with fractional DNA content.

Keywords:

  • cell cycle;
  • DNA ploidy;
  • DNA index;
  • Hoechst 33342;
  • propidium iodide;
  • PI;
  • 4′6-diamidino-2-phenylindole;
  • DAPI