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Related Experiment Videos

A simple technique for quantifying apoptosis in 96-well plates.

Deborah Ribble1, Nathaniel B Goldstein, David A Norris

  • 1Dermatology Department, UCHSC at Fitzsimons, PO Box 6511, Aurora, CO 80045, USA. Deborah.Ribble@uchsc.edu

BMC Biotechnology
|May 12, 2005
PubMed
Summary

A modified ethidium bromide and acridine orange (EB/AO) staining assay offers a faster, gentler method for quantifying apoptosis in both live and dead cells. This 96-well plate assay provides comparable results to conventional methods, improving cell viability analysis.

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Area of Science:

  • Cell Biology
  • Biochemistry

Background:

  • Quantifying apoptosis is crucial in biological studies.
  • Current methods for apoptosis quantification have limitations including multiple steps, cell damage, and non-specific detection.
  • A modified ethidium bromide and acridine orange (EB/AO) staining assay in a 96-well plate format is presented to overcome these limitations.

Purpose of the Study:

  • To develop a modified EB/AO staining assay for apoptosis quantification in a 96-well plate format.
  • To overcome the limitations of existing apoptosis detection methods.
  • To combine the advantages of the 96-well format with the conventional EB/AO method.

Main Methods:

  • A modified ethidium bromide and acridine orange (EB/AO) staining assay was developed.
  • The assay was performed entirely within a 96-well plate.

Related Experiment Videos

  • The modified method was compared to the conventional EB/AO method using suspension (Jurkat) and adherent (A375) cells.

    • Main Results:

    • The modified EB/AO assay achieved quantification results comparable to the conventional EB/AO method.
    • Comparable results were observed for both suspension and adherent cells.
    • The new method demonstrated effectiveness under normal growth and apoptosis-inducing conditions.

    Conclusions:

    • The modified EB/AO assay significantly reduces assay time by eliminating detaching and washing steps.
    • The method minimizes damage to adherent cells and reduces the loss of floating cells.
    • This improved assay is particularly beneficial for quantifying apoptosis in adherent cells.