一种敏感的辣根过氧化物酶反应法:葡萄糖氧化酶-二氨基联苯胺-硫酸镍铵法

THE GLUCOSE OXIDASE-DAB-NICKEL METHOD IN PEROXIDASE HISTOCHEMISTRY OF THE NERVOUS SYSTEM

神经系统束路追踪法或免疫组化法的最后步骤,常常需要用辣根过氧化物酶的呈色来显示。显示辣根过氧化物酶最常用的色素为二氨基联苯胺,其呈色产物为棕色,稳定,而且电子密度高,适于作光镜和电镜研究。但该法不够敏感,不能显示低浓度的过氧化物酶是其缺点。我们用葡萄糖氧化酶和葡萄糖代替双氧水供氧,并用镍盐增强二氨基联苯胺的呈色强度,得出一种黑色反应产物,比棕色产物的二氨基联苯胺原法及其他二氨基联苯胺法均敏感。能显示免疫反应阳性的细微结构,对显示阳性反应的纤维和终末效果极佳。并可与二氨基联苯胺原法或其他法合用作免疫组化双标记或免疫组化与束路追踪双标记。

A glucose oxidase-3,3′ diaminobenzidine-nickel method was developed.This technique can successfully demonstrate the details of the immunoreactive structures and PHA-L labeled cell bodies and their processes.It is especially beneficial for visualizing fibers and terminals.It is more sensitive than the regular 3,3′ diamino- benzidine method and the glucose oxidase-3,3′ diaminobenzidine technique,and very stabilized.