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Related Experiment Videos

Apoptosis assessment by the DNA diffusion assay.

Narendra P Singh1

  • 1Department of Bioengineering, University of Washington, Seattle, USA.

Methods in Molecular Medicine
|May 25, 2005
PubMed
Summary

This study introduces a simple DNA diffusion assay to quantify apoptosis. The method detects DNA fragmentation in apoptotic cells, visualized as a halo using YOYO-1 staining.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Apoptosis, or programmed cell death, is a crucial biological process.
  • Quantifying apoptosis is essential for understanding various physiological and pathological conditions.
  • Existing methods for apoptosis detection can be complex or lack sensitivity.

Purpose of the Study:

  • To develop a simple, sensitive, and reliable DNA diffusion assay for quantifying apoptosis.
  • To validate the assay for detecting apoptosis in human leukocytes.

Main Methods:

  • The assay utilizes the principle of DNA diffusion from apoptotic cells with alkali-labile sites.
  • Cells are embedded in agarose microgels, lysed under alkaline conditions, and DNA is precipitated.
  • DNA is stained with the fluorescent dye YOYO-1 for visualization.

Main Results:

  • Apoptotic cells exhibit a characteristic halo formation due to DNA diffusion.
  • The assay successfully detected and quantified apoptosis in human leukocytes (neutrophils).
  • The DNA diffusion assay provides a clear visual indicator of apoptotic cells.

Conclusions:

  • The developed DNA diffusion assay is a simple, sensitive, and reliable method for apoptosis quantification.
  • This assay offers a valuable tool for researchers studying apoptosis in various cell types.
  • The halo appearance serves as a distinct marker for identifying apoptotic cells.

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