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Related Experiment Videos

An enhanced method for post-embedding immunocytochemical staining which preserves cell membranes.

M A Berryman1, R D Rodewald

  • 1Department of Biology, University of Virginia, Charlottesville 22901.

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
|February 1, 1990
PubMed
Summary

A new immunogold staining method preserves cell ultrastructure and antigenicity in plastic-embedded tissues. This technique successfully localized immunoglobulin G receptors in rat jejunal epithelium.

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Effects of tannic acid on antigenicity and membrane contrast in ultrastructural immunocytochemistry.

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society·1992
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Area of Science:

  • Cell Biology
  • Immunocytochemistry
  • Microscopy

Background:

  • Conventional immunogold staining methods often compromise cell ultrastructure.
  • Preserving antigenicity and ultrastructure is crucial for accurate localization studies.

Purpose of the Study:

  • To develop an improved immunogold staining technique for plastic-embedded tissues.
  • To achieve high specificity without sacrificing cellular integrity.
  • To localize the immunoglobulin G receptor in neonatal rat jejunal epithelium.

Main Methods:

  • Utilized a combination of optimized standard techniques including specific primary fixation, uranyl acetate post-fixation, acetone dehydration, and low-temperature embedding in LR Gold resin.
  • Employed osmium tetroxide staining after immunogold labeling for thin sections.

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  • Used monoclonal primary antibodies and colloidal gold-labeled secondary antibodies for staining.

    • Main Results:

    • Achieved high levels of specific immunogold staining.
    • Successfully preserved excellent cell ultrastructure, including membrane integrity.
    • Resolved the immunoglobulin G receptor within membrane compartments involved in transport.

    Conclusions:

    • The developed method provides high-quality immunogold staining while maintaining ultrastructural preservation.
    • This technique is suitable for localizing membrane receptors and applicable to various immunocytochemical studies.
    • The method offers a valuable tool for ultrastructural immunolocalization in diverse biological systems.