Resin Embedment of Organs and Postembedment Localization of Antigens by Immunoperoxidase Methods

Overview

Journal Histochemistry

Specialty Biochemistry

Date 1981 Jan 1

PMID 6170617

Citations 6

Authors

W D Kuhlmann

R Krischan

Affiliations

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Abstract

Various procedures for nonpolar and polar resin embedment were applied to mouse and rat livers for the study of postembedment immunolocalization of alpha 1-fetoprotein, albumin and the microsomal enzyme epoxide hydrolase. Fixations with formaldehyde and with formaldehyde-glutaraldehyde mixtures were used for tissue stabilization. Both fixation schedules did not abolish immunoreactivity. Treatment of liver with inert compounds such as polyvinylpyrrolidones or chemical modification of antigens with ethyl acetimidate prior to embedment improved immuno-staining. Either the low-polarity solvent ethanol or the highly polar ethylene glycol could be employed as dehydrating agents. Antigens were readily localized in sections from Epon 812 embedded livers. For this purpose, polymerized resin had to be partially removed. On the other hand, immunoreactivity of antigens was only faint after embedment in an epoxy-resin based on diepoxide octane. Also, antigens reacted faintly in sections from livers which were embedded at 0 degrees C in the polar acrylate-methacrylate based Lowicryl K4M resin. The indirect peroxidase labelled antibody method was as specific and sensitive as the PAP technique. Optimal antigen detection was attained with antibodies isolated by affinity chromatography and purified peroxidase conjugates. Apart from purified immunological reagents, the addition of high molarity sodium chloride and bovine serum albumin to the wash solutions enhanced immunohistological specificity.

Citing Articles

Hepatic progenitor cells, stem cells, and AFP expression in models of liver injury.

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Advances in ultrastructural postembedment localization of antigens in Epon sections with peroxidase labelled antibodies.

Kuhlmann W, Peschke P Histochemistry. 1982; 75(2):151-61.

PMID: 6752110 DOI: 10.1007/BF00496006.

Standardization of various applications of methacrylate embedding and silver methenamine for light and electron microscopy immunocytochemistry.

Rodriguez E, Yulis R, Peruzzo B, Alvial G, Andrade R Histochemistry. 1984; 81(3):253-63.

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The distribution of cell surface glycoconjugates during mouse secondary neurulation.

Griffith C, Wiley M Anat Embryol (Berl). 1989; 180(6):567-75.

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Light microscopical detection of antigens and lectin binding sites with gold-labelled reagents on semi-thin Lowicryl K4M sections: usefulness of the photochemical silver reaction for signal amplification.

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