锇固定后用4-氯-1-萘酚对胰岛素进行电子显微镜免疫过氧化物酶染色。

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

作者信息

Baskin D G, Mar H, Gorray K C, Fujimoto W Y

出版信息

J Histochem Cytochem. 1982 Jul;30(7):710-2. doi: 10.1177/30.7.6179987.

DOI:10.1177/30.7.6179987

Abstract

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.

摘要

利用底物溶液中含4-氯-1-萘酚（CN）的免疫过氧化物酶法，在锇固定后完成了豚鼠胰腺B细胞中胰岛素的超微结构定位。该方案的主要特点是：a）四氧化锇后固定；b）省略过氧化氢“蚀刻”；c）底物溶液中使用CN代替二氨基联苯胺；d）底物反应后去除四氧化锇；e）醋酸铀和柠檬酸铅复染。该方法使用高度稀释的抗血清产生低背景的强烈特异性染色，似乎可用于锇固定组织中多种肽抗原的包埋后免疫过氧化物酶染色。