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Related Experiment Videos

Improved cytochemical methods for demonstrating cell death using LR White as an embedding medium.

A A Al-Hazzaa1, I D Bowen

  • 1School of Pure and Applied Biology, University of Wales Cardiff, UK.

The Histochemical Journal
|April 1, 1999
PubMed
Summary
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New techniques using LR White resin enable precise localization of DNA fragmentation, a key marker of apoptosis, in cancer cells. These methods allow detailed comparison at both light and electron microscope levels for better understanding of cell death.

Area of Science:

  • Cell Biology
  • Histopathology
  • Microscopy

Background:

  • Apoptosis is programmed cell death, crucial for development and disease.
  • DNA fragmentation is a hallmark of apoptosis.
  • Accurate localization of DNA fragmentation is vital for understanding apoptosis.
  • Current methods may have limitations in resolution or comparability.

Purpose of the Study:

  • To describe novel techniques for in situ DNA fragmentation localization in apoptosis.
  • To enable simultaneous light and electron microscopic analysis.
  • To compare different detection methods for nucleosomal histones.

Main Methods:

  • Utilized LR White resin embedding for tissue preservation.
  • Employed anti-histone antibody for detecting DNA fragmentation.

Related Experiment Videos

  • Applied 3,3'-diaminobenzidine (DAB) and silver-enhanced techniques for light microscopy.
  • Used colloidal gold-based anti-histone technique for electron microscopy.
  • Investigated untreated squamous cell carcinoma of the pharynx.
  • Performed parallel histochemical localization of acid phosphatase activity.

    • Main Results:

    • Successfully localized DNA fragmentation characteristic of apoptosis at both light and electron microscope levels.
    • Enabled direct comparison between light microscopic (DAB, silver-enhanced) and electron microscopic (colloidal gold) histone localization.
    • Demonstrated the utility of LR White resin techniques for apoptosis research.
    • Provided parallel data on acid phosphatase activity.

    Conclusions:

    • The described LR White resin-based techniques are effective for in situ apoptosis detection.
    • These methods facilitate high-resolution comparative analysis of DNA fragmentation and histone exposure.
    • The techniques are applicable to various tissues, including squamous cell carcinoma.
    • This work advances the understanding of apoptosis mechanisms through improved visualization.