Unit

UNIT 7.2 Critical Aspects in Analysis of Cellular DNA Content

  1. Zbigniew Darzynkiewicz

Published Online: 1 APR 2011

DOI: 10.1002/0471142956.cy0702s56

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Darzynkiewicz, Z. 2011. Critical Aspects in Analysis of Cellular DNA Content. Current Protocols in Cytometry. 56:7.2:7.2.1–7.2.8.

Author Information

  1. The Brander Cancer Research Institute and Department of Pathology, New York Medical Center, Valhalla, New York

Publication History

  1. Published Online: 1 APR 2011
  2. Published Print: APR 2011

Abstract

This unit covers general aspects of DNA content analysis and provides introductory or complementary information to the specific protocols of DNA content assessment in this chapter. It describes principles of DNA content analysis and outlines difficulties and pitfalls common to these methods. It also reviews methods of DNA staining in live, permeabilized, and fixed cells, and in cell nuclei isolated from paraffin-embedded tissues, as well as the approaches to stain DNA concurrently with cell immunophenotype. This unit addresses factors affecting accuracy of DNA measurement, such as chromatin features restricting accessibility of fluorochromes to DNA, stoichiometry of interaction with DNA, and “mass action law” characterizing binding to DNA in relation to unbound fluorochrome concentration. It also describes controls to ensure accuracy and quality control of DNA content determination and principles of DNA ploidy assessment. Because many aspects of DNA content analysis are common to protocols in units 7.3, 7.6, 7.16, 7.20, 7.23, & 7.25, certain parts of this unit provide information redundant with commentaries in these units. Curr. Protoc. Cytom. 56:7.2.1-7.2.8. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • cell cycle;
  • apoptosis;
  • ploidy;
  • DNA index;
  • stoichiometry;
  • fluorochrome;
  • chromatin