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Immunohistochemistry of Brain Tissues
Akira Hara1, Toshiaki Taniguchi2, Tomohiro Kanayama2
1Department of Tumor Pathology, Gifu University Graduate School of Medicine, Gifu, Japan. hara.akira.y7@f.gifu-u.ac.jp.
Insights
Immunohistochemistry (IHC) is a vital technique for analyzing proteins in brain tissues using formalin-fixed paraffin-embedded samples. Standardized methods and automated immunostainers enhance the accuracy of this crucial pathological analysis.
Area of Science:
- Neuroscience
- Pathology
- Biochemistry
Background:
- Immunohistochemistry (IHC) is fundamental for histological and pathological analysis.
- It enables protein detection and characterization in diverse tissues, notably brain tissues.
- Formalin-fixed paraffin-embedded (FFPE) tissues are commonly used due to their stability and versatility.
Purpose of the Study:
- To detail standard immunohistochemistry methods for brain slice sections.
- To highlight IHC's role in analyzing neuronal and glial cell localization and function.
- To introduce automated immunostainers for standardized IHC.
Main Methods:
- Standard immunohistochemistry protocols applied to brain tissue sections.
- Chromogenic IHC utilizing enzyme-conjugated antibodies and substrates.
- Fluorescent IHC employing fluorescently tagged antibodies.
Main Results:
- Demonstration of standard IHC techniques for brain tissue analysis.
- Comparison of chromogenic and fluorescent visualization methods.
- Presentation of an automated immunostainer for reproducible results.
Conclusions:
- IHC is essential for detailed analysis of brain tissue components.
- Standardized methods, including automated options, improve the reliability of IHC.
- Accurate protein characterization via IHC aids in understanding neural cell function and pathology.
Abstract:
Immunohistochemistry (IHC) is the basis of histological or pathological analysis and is widely used to enable the detection and characterization of proteins in various organ tissues, including brain tissues. IHC is commonly performed on formalin-fixed paraffin-embedded (FFPE) tissues because of their easy storage and versatility. IHC is a key method for providing more accurate analysis of localization and function of neurons, neuroendocrine cells, and neural stem cells in the brain and other nervous systems. The related cells such as glial cells and neurovascular units have also been analyzed by IHC. Visualization of antibody-antigen interactions can be performed primarily in one of the following ways: chromogenically stained IHC and fluorescently stained IHC. In chromogenically stained IHC, an antibody is chemically conjugated to an enzyme, such as peroxidase, that can be reacted with a suitable substrate to give a colored product. In fluorescently stained IHC, the antibodies are finally tagged with fluorescent chemicals such as fluorescein isothiocyanate (FITC) or rhodamine. Here, we describe the standard methods of IHC applied to brain slice sections. Furthermore, an automated immunostainer is presented as another option for standardized immunohistochemistry.
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