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A universal post-embedding protocol for immunogold labelling of osmium-fixed, epoxy resin-embedded tissue
1Department of Cell Biology, Neurobiology and Anatomy, College of Medicine, University of Cincinnati, OH 45267, USA.
Journal of Electron Microscopy
|January 1, 1997
Summary
Double etching epoxy-embedded sections with 3% sodium metaperiodate offers effective epitope expression. This method avoids specimen bleaching and resin damage seen with more concentrated solutions.
Area of Science:
- Immunohistochemistry
- Electron Microscopy Techniques
Background:
- Epitope retrieval is crucial for antigen detection in epoxy-embedded ultrathin sections.
- Sodium metaperiodate is a common oxidizing agent for enhancing epitope exposure.
Purpose of the Study:
- To evaluate the efficacy of double etching with 3% sodium metaperiodate for epitope expression.
- To compare this method with saturated and half-saturated sodium metaperiodate solutions.
- To assess potential side effects like bleaching and resin damage.
Main Methods:
- Epoxy-embedded ultrathin sections were double etched using 3% sodium metaperiodate.
- Epitope expression was compared to sections treated with saturated and half-saturated sodium metaperiodate.
- Specimen integrity (bleaching, holes) was assessed.
Main Results:
- Double etching with 3% sodium metaperiodate achieved epitope expression comparable to more concentrated solutions.
- This method did not cause specimen bleaching.
- No holes were generated in the plastic resin.
Conclusions:
- Double etching with 3% sodium metaperiodate is a viable and gentler alternative for epitope retrieval.
- This technique preserves specimen integrity while ensuring adequate antigen accessibility for analysis.