An improved immunocytochemical staining method for large semi-thin plastic epon sections: application to GABA in rat cerebral cortex. | Zendy

H.J. Romijn | Zendy; A.W.J.W. Janszen | Zendy; C.W. Pool | Zendy; Ruud M. Buijs | Zendy

Open Access

An improved immunocytochemical staining method for large semi-thin plastic epon sections: application to GABA in rat cerebral cortex.

Author(s) -

H.J. Romijn,

A.W.J.W. Janszen,

C.W. Pool,

Ruud M. Buijs

Publication year - 1993

Publication title -

journal of histochemistry and cytochemistry

Language(s) - English

Resource type - Journals

eISSN - 1551-5044

pISSN - 0022-1554

DOI - 10.1177/41.8.7687264

Subject(s) - staining , immunocytochemistry , glutaraldehyde , neurotransmitter , reproducibility , immunohistochemistry , cerebral cortex , biology , chemistry , biomedical engineering , microbiology and biotechnology , pathology , neuroscience , chromatography , central nervous system , medicine , immunology , endocrinology

We describe here a procedure that significantly enhances the intensity and method-specificity of immunocytochemical staining in large mounted semi-thin plastic Epon sections. The procedure was developed for the detection of the neurotransmitter gamma-aminobutyric acid (GABA) in rat brain tissue fixed with glutaraldehyde, but it may also be helpful in unmasking other antigens under different conditions. In addition to some practical suggestions for improving the reproducibility of the staining procedure, we demonstrate that the crucial step in the procedure is pre-treatment of the deplasticized sections with proteinase-K before exposure to the first antibody. This leads to a high morphological resolution and an excellent immunocytochemical signal.

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