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Electron microscopic immunoperoxidase staining of insulin using 4- chloro-1-naphthol after osmium fixation

DG Baskin, H Mar, KC Gorray and WY Fujimoto

Ultrastructural localization of insulin in B cells of guinea pig pancreas was accomplished after osmium fixation with an immunoperoxidase procedure that utilized 4-chloro-1-naphthol (CN) in the substrate solution. The principal features of this protocol were: a) osmium tetroxide postfixation; b) omission of hydrogen peroxide "etching"; c) use of CN instead of diaminobenzidine in the substrate solution; d) elimination of osmium tetroxide after the substrate reaction; e) uranyl acetate and lead citrate counterstaining. This procedure produces intense specific staining with low background using highly dilute antiserum, and appears to be useful for postembedding immunoperoxidase staining of a variety of peptide antigens in osmium- fixed tissue.

Volume 30, Issue 7, pp. 710-712, 07/01/1982
Copyright © 1982 by The Histochemical Society


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The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 1982