Critical Aspects in Analysis of Cellular DNA Content
Open Access
- 1 April 2011
- journal article
- unit
- Published by Wiley in Current Protocols in Cytometry
- Vol. 56 (1), 7.2.1-7.2.8
- https://doi.org/10.1002/0471142956.cy0702s56
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
This publication has 26 references indexed in Scilit:
- Induction of DNA damage response by the supravital probes of nucleic acidsCytometry Part A, 2009
- High‐resolution multi‐parameter DNA flow cytometry enables detection of tumour and stromal cell subpopulations in paraffin‐embedded tissuesThe Journal of Pathology, 2005
- Cytometry of the cell cycle: Cycling through historyCytometry Part A, 2004
- Simultaneous flow cytometric analysis of two cell surface markers, telomere length, and DNA contentCytometry, 2002
- Laser scanning cytometry distinguishes lymphocytes, monocytes, and granulocytes by differences in their chromatin structureCytometry, 1997
- Cytometry in cell necrobiology: Analysis of apoptosis and accidental cell death (necrosis)Cytometry, 1997
- An improved hedley method for preparation of paraffin-embedded tissues for flow cytometric analysis of ploidy and S-phaseCytometry, 1991
- Effect of drug efflux blockers on vital staining of cellular DNA with hoechst 33342Cytometry, 1987
- Changes in accessibility of DNA to various fluorochromes during spermatogenesisCytometry, 1986
- Simultaneous quantitation of hoechst 33342 and immunofluorescence on viable cells using a fluorescence activated cell sorterCytometry, 1980