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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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Updated: Jun 17, 2026

Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids

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Cell fixatives for immunostaining.

Maria Célia Jamur¹, Constance Oliver

¹Department of Cell and Molecular Biology, Faculdade de Medicina de Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.

Methods in Molecular Biology (Clifton, N.J.)

|December 17, 2009

Summary

This summary is machine-generated.

Fixation is crucial for immunostaining, balancing tissue preservation with antigenicity. This review covers chemical fixation methods for light microscopy, detailing agents like formaldehyde and methanol.

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Area of Science:

Histology
Immunohistochemistry
Microscopy

Background:

Fixation is a critical step in immunostaining procedures.
The primary goals of fixation are to preserve tissue morphology and maintain antigenicity.
Various methods exist, including chemical and physical approaches, each with specific applications.

Purpose of the Study:

To provide a comprehensive overview of chemical fixation techniques.
To detail the mechanisms and applications of common chemical fixatives.
To focus on methods suitable for light microscopy.

Main Methods:

Discussion of chemical fixation agents, including cross-linking agents (e.g., formaldehyde, glutaraldehyde) and protein-precipitating solvents (e.g., acetone, methanol).
Comparison of chemical fixation with physical methods like freezing and air-drying.
Emphasis on techniques relevant to light microscopy applications.

Main Results:

Chemical fixation offers diverse options for balancing morphological and antigenic preservation.
Cross-linking agents effectively preserve structure by forming molecular bridges.
Solvents rapidly precipitate proteins, aiding in cellular detail preservation.

Conclusions:

Chemical fixation is essential for successful immunostaining, requiring careful selection of methods.
Understanding the properties of different fixatives is key to optimizing experimental outcomes.
This chapter highlights chemical fixation strategies vital for light microscopy.