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Related Experiment Videos

A rapid cell membrane permeability test using fluorescent dyes and flow cytometry.

M Aeschbacher1, C A Reinhardt, G Zbinden

  • 1Institute of Toxicology, Federal Institute of Technology, Schwerzenbach, Switzerland.

Cell Biology and Toxicology
|June 1, 1986
PubMed
Summary

This study introduces a rapid cell membrane integrity assay using fluorescein diacetate and ethidium bromide. The test effectively quanties chemical effects on cell membranes, detecting damage before viability loss.

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Introduction.

Toxicology in vitro : an international journal published in association with BIBRA·2010

Area of Science:

  • Biochemistry
  • Cell Biology
  • Toxicology

Background:

  • Cell membrane integrity is crucial for cell viability.
  • Assessing chemical effects on cell membranes requires reliable and rapid methods.
  • Existing viability assays may not detect early membrane damage.

Purpose of the Study:

  • To develop and validate a rapid, reliable assay for detecting cytotoxic chemicals that impact cell membranes.
  • To quantify concentration-dependent effects of chemicals on cell membrane integrity.
  • To monitor early chemical insults to cell membranes preceding loss of viability.

Main Methods:

  • Utilized a dual-staining technique with fluorescein diacetate (FDA) and ethidium bromide (EB) on rat thymocytes.
  • Employed flow cytometry for two-parameter analysis of dye retention and exclusion.

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  • Quantified chemical effects by measuring changes in FDA-fluorescence and EB-staining relative to controls.

    • Main Results:

    • Intact cells fluoresced green (cytoplasm) with FDA, while membrane-damaged cells fluoresced red (nucleus/RNA) with EB.
    • The assay detected concentration-dependent effects of detergents and solvents on cell membranes.
    • Measured dye retention (FDA) and dye exclusion (EB) changes accurately reflected chemical insult.

    Conclusions:

    • The combined FDA/EB assay provides a sensitive and rapid method for assessing cell membrane damage.
    • This assay can monitor early chemical insults to cell membranes, preceding traditional viability measurements.
    • The technique offers a valuable tool for toxicological screening and research on membrane-active compounds.