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Protocol for HER2 FISH Using a Non-cross-linking, Formalin-free Tissue Fixative to Combine Advantages of Cryo-preservation and Formalin Fixation
Published on: December 25, 2017
Tissue preparation for histochemistry: fixation, embedding, and antigen retrieval for light microscopy
Tim D Hewitson1, Belinda Wigg, Gavin J Becker
1Department of Nephrology, The Royal Melbourne Hospital, Melbourne, VIC, Australia.
Methods in Molecular Biology (Clifton, N.J.)
|December 5, 2009
Summary
Histochemistry uses specialized assays on tissue samples for in situ biological study. Proper tissue preparation, including freezing or paraffin embedding, is crucial for accurate results and antigen preservation.
Area of Science:
- Histochemistry
- Immunohistochemistry
- Molecular Biology Assays
Background:
- Histochemical techniques enable in situ study of tissue and organ biology.
- Success relies heavily on appropriate tissue preparation methods.
Purpose of the Study:
- To outline essential tissue preparation steps for routine histochemical analysis.
- To compare frozen and paraffin-embedded tissue preparation for histological analysis.
Main Methods:
- Describing basic tissue preparation for histochemical analysis.
- Discussing frozen tissue preparation for minimal epitope alteration.
- Detailing paraffin embedding for enhanced structural integrity.
Main Results:
- Frozen tissue may offer superior staining due to less epitope alteration.
- Paraffin embedding provides better morphology but can reduce antigenicity.
- Antigen retrieval techniques can mitigate antigen loss in paraffin-embedded tissues.
Conclusions:
- Adequate tissue preparation is fundamental for successful histochemical analysis.
- Choosing between frozen and paraffin-embedded methods depends on balancing morphological detail and antigen preservation.
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