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Fixation and Sectioning

Fixation and Sectioning

Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...

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Preparing Paraffin Tissue Sections for Staining.

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|March 2, 2021

Summary

This summary is machine-generated.

Formalin-fixed, paraffin-embedded (FFPE) tissues are common in histology. Heat-induced epitope retrieval can improve antibody-antigen binding by unmasking epitopes masked during FFPE processing.

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Published on: December 25, 2017

Area of Science:

Histology
Immunohistochemistry
Biotechnology

Background:

Formalin-fixed, paraffin-embedded (FFPE) tissues are widely used in histological studies.
An extensive atlas of tissues and organs exists from FFPE samples, making them valuable for comparative analysis.
Harsh FFPE processing can degrade antigens and mask epitopes, hindering antibody-antigen binding.

Purpose of the Study:

To describe the procedures for preparing paraffin tissue sections.
To address the challenge of poor antigen preservation in FFPE samples.
To present a method for improving antibody-antigen binding in FFPE tissue sections.

Main Methods:

Standard histological fixation and embedding procedures for paraffin tissue sections.
Sensitive detection methods and multi-layer amplification techniques for cell staining.
Heat-induced epitope retrieval (HIER) to reverse protein cross-linking and unmask epitopes.

Main Results:

FFPE processing conditions can lead to antigen degradation and epitope masking.
Sensitive detection and amplification methods are often required for FFPE tissue staining.
Heat-induced epitope retrieval effectively unmasks epitopes, enhancing antibody-antigen binding.

Conclusions:

FFPE tissue processing presents challenges for antigen preservation and antibody binding.
Heat-induced epitope retrieval is a valuable technique for improving immunohistochemical staining of FFPE tissues.
This method facilitates more informative antigen localization studies by overcoming fixation-induced limitations.