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使用铈离子和水性孵育介质通过光学显微镜检测产生过氧化氢的氧化酶。

Light microscopical detection of H2O2-generating oxidases using cerium ions and aqueous incubation media.

作者信息

Gossrau R, Frederiks W M, van Noorden C J, Klebe S, Ruhnke M

机构信息

Department of Anatomy, Free University of Berlin, Germany.

出版信息

Acta Histochem. 1991;90(1):27-37. doi: 10.1016/S0065-1281(11)80153-4.

Abstract

Light microscopical procedures were optimized and tested for specificity for the histochemical demonstration of D-amino acid oxidase, alpha-hydroxy acid oxidase, monoamine oxidase, and xanthine oxidase using cerium ions and a visualization step originally described by Angermüller and Fahimi (1988a, b), and modified for D-amino acid oxidase by Gossrau et al. (1989). The visualization medium contained diaminobenzidine, cobalt ions, and small amounts of hydrogen peroxide. Procedures of pretreatment of cryostat sections, types of substrate, concentrations of substrates, and cerium ions were varied. Optimum procedures are reported for the detection of these oxidases in different rat tissues. The results are compared with those obtained with other methods described for the demonstration of hydrogen peroxide-generating oxidases such as the tetrazolium, Hatchett Brown, and coupled peroxidatic methods.

摘要

对光学显微镜方法进行了优化,并测试了其特异性,用于使用铈离子以及最初由安格穆勒和法希米(1988a,b)描述并由戈斯劳等人(1989)针对D-氨基酸氧化酶进行修改的可视化步骤,对D-氨基酸氧化酶、α-羟基酸氧化酶、单胺氧化酶和黄嘌呤氧化酶进行组织化学示踪。可视化介质包含二氨基联苯胺、钴离子和少量过氧化氢。对冷冻切片的预处理程序、底物类型、底物浓度和铈离子进行了改变。报告了在不同大鼠组织中检测这些氧化酶的最佳程序。将结果与使用其他用于证明产生过氧化氢的氧化酶的方法(如四唑盐法、哈奇特·布朗法和偶联过氧化物酶法)所获得的结果进行了比较。

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