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Related Experiment Videos

Ethidium bromide: a nucleic acid stain for tissue section.

W A Franklin, J D Locker

    The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
    |April 1, 1981
    PubMed
    Summary
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    Ethidium bromide (EB) is a fluorescent dye that stains cell nuclei in tissue sections. It can also differentiate DNA and RNA when combined with enzyme digestion, aiding in cellular localization.

    Area of Science:

    • Molecular Biology
    • Cell Biology
    • Histology

    Background:

    • Ethidium bromide (EB) is a phenanthridinium dye known for its selective intercalation into double-stranded nucleic acids.
    • EB exhibits a significant and specific increase in fluorescence upon binding to DNA.
    • Its application in staining fixed tissue sections allows for high-resolution visualization of microscopic details.

    Purpose of the Study:

    • To evaluate ethidium bromide as a stain for fixed tissue sections in fluorescence microscopy.
    • To assess the utility of EB as a counterstain in immunofluorescence.
    • To explore the potential of EB in combination with enzymatic digestion for differential localization of DNA and RNA.

    Main Methods:

    • Staining of fixed tissue sections with ethidium bromide.

    Related Experiment Videos

  • Observation of fluorescence microscopy to assess nuclear and cytoplasmic staining.
  • Enzymatic digestion of tissue sections using trypsin and ribonuclease.
  • Counterstaining with fluorescein for immunofluorescence applications.

    • Main Results:

    • Ethidium bromide effectively stains cellular nuclei with excellent resolution.
    • Intense cytoplasmic staining observed in some tissues (e.g., pancreatic acini) was eliminated by trypsin and ribonuclease digestion.
    • The orange fluorescence of EB is distinct from the green fluorescence of fluorescein, making it a suitable counterstain.
    • Combined enzymatic digestion allowed for differential localization of DNA and RNA.

    Conclusions:

    • Ethidium bromide is a practical and effective stain for fluorescence microscopy of tissue sections.
    • EB serves as an excellent counterstain for immunofluorescence.
    • The combination of EB staining with enzymatic digestion offers a simple method for differentiating DNA and RNA within tissues.