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Simple, Affordable, and Modular Patterning of Cells using DNA
Published on: February 24, 2021
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Analysis of Cellular DNA Content by Flow Cytometry.
Zbigniew Darzynkiewicz1, Xuan Huang2, Hong Zhao1
1Department of Pathology and Brander Cancer Research Institute, New York Medical College, Valhalla, New York.
Current Protocols in Cytometry
|October 3, 2017
Summary
Flow cytometry offers robust methods for analyzing cellular DNA content. This technique reveals cell cycle distribution, apoptosis frequency, and DNA ploidy, aiding in cell biology research.
Area of Science:
- Cell Biology
- Molecular Biology
- Biotechnology
Background:
- Accurate measurement of cellular DNA content is crucial for understanding cell proliferation, apoptosis, and ploidy.
- Flow cytometry is a powerful technique for quantitative analysis of cellular components, including DNA.
Purpose of the Study:
- To present universally applicable methods for measuring cellular DNA content using flow cytometry.
- To detail techniques for cell permeabilization and DNA staining for accurate analysis.
- To describe algorithms for DNA content histogram deconvolution.
Main Methods:
- Staining of fixed cells using detergents and/or proteolytic treatments for DNA accessibility.
- Supravital cell staining with Hoechst 33342 for live cell sorting based on DNA content.
- Methods for staining cell nuclei isolated from paraffin-embedded tissues.
Main Results:
- Established simple and universally applicable methods for DNA content analysis.
- Provided protocols for both fixed and live cell staining, as well as for tissue samples.
- Listed available algorithms for histogram deconvolution to quantify cell cycle phases and apoptosis.
Conclusions:
- Flow cytometry provides versatile methods for assessing cell cycle distribution, apoptosis, and DNA ploidy.
- The presented techniques are applicable to various cell types and sample preparations, including fixed cells, live cells, and paraffin-embedded tissues.
- Computational algorithms are essential for accurate interpretation of DNA content histograms.
