NYMC Faculty Publications

Analysis of Cellular DNA Content by Flow Cytometry

DOI

10.1002/cpim.36

Journal Title

Current Protocols in Immunology

First Page

7.5.1

Last Page

7.5.20

Document Type

Article

Publication Date

11-1-2017

Department

Pathology, Microbiology and Immunology

Keywords

DNA Ploidy, Cell Cycle, DNA Fragmentation, Apoptosis, Ploidies, Flow Cytometry

Disciplines

Biological Phenomena, Cell Phenomena, and Immunity | Medical Cell Biology | Medicine and Health Sciences | Nanotechnology

Abstract

Cellular DNA content can be measured by flow cytometry with the aim of : (1) revealing cell distribution within the major phases of the cell cycle, (2) estimating frequency of apoptotic cells with fractional DNA content, and/or (3) disclosing 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/or proteolytic treatment to permeabilize cells and make DNA accessible to fluorochrome. Additionally, supravital cell staining with Hoechst 33342, which is primarily used for sorting live cells based on DNA-content differences for their subsequent culturing, is described. Also presented are methods for staining cell nuclei isolated from paraffin-embedded tissues. Available algorithms are listed for deconvolution of DNA-content-frequency histograms to estimate percentage of cells in major phases of the cell cycle and frequency of apoptotic cells with fractional DNA content.

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