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Published on: December 13, 2017
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.
Insights
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.
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.
Abstract:
Fixation is one of the most critical steps in immunostaining. The object of fixation is to achieve good morphological preservation, while at the same time preserving antigenicity. Tissue blocks, sections, cell cultures or smears are usually immersed in a fixative solution, while in other situations, whole body perfusion of experimental animals is preferable. Fixation can be accomplished by either chemical or physical methods. The chemical methods include cross-linking agents such as formaldehyde, glutaraldehyde and succinimide esters as well as solvents such as acetone and methanol, which precipitate proteins. Of the physical methods, freezing tissue and air drying are most widely used. This chapter deals with the chemical fixation methods most commonly used for light microscopy.
