利用3,3'-二氨基联苯胺在光学和电子显微镜下进行过氧化物酶体的细胞化学检测

Cytochemical Detection of Peroxisomes in Light and Electron Microscopy with 3,3'-diaminobenzidine.

作者信息

Fahimi H Dariush

机构信息

Division of Medical Cell Biology, Department of Anatomy and Cell Biology, University of Heidelberg, Im Neuenheimer Feld 307, D-69120, Heidelberg, Germany.

出版信息

Methods Mol Biol. 2017;1595:93-100. doi: 10.1007/978-1-4939-6937-1_10.

DOI:10.1007/978-1-4939-6937-1_10

PMID:28409455

Abstract

Peroxisomes are ubiquitous dynamic and multifunctional organelles that contribute to numerous anabolic and catabolic pathways, being essential for human health and development. Their best known functions include the oxidation of fatty acids and metabolism of hydrogen peroxide with catalase as a marker enzyme. Indeed, historically, it was the cytochemical staining of catalase in many different cells and tissues that revealed the ubiquitous presence of peroxisomes in almost all animal and plant cells. In this chapter, the method for cytochemical staining of catalase with the alkaline 3, 3'-diaminobenzidine (DAB) is described. Since aldehyde fixation is a prerequisite for staining of catalase with DAB, a method for perfusion fixation of rat liver with glutaraldehyde is presented prior to the cytochemical staining method and the subsequent tissue processing for light and electron microscopy.

摘要

过氧化物酶体是普遍存在的、动态的多功能细胞器，参与众多合成代谢和分解代谢途径，对人类健康和发育至关重要。它们最广为人知的功能包括脂肪酸氧化以及以过氧化氢酶作为标记酶的过氧化氢代谢。实际上，从历史角度来看，正是在许多不同细胞和组织中对过氧化氢酶进行细胞化学染色，才揭示了过氧化物酶体几乎在所有动植物细胞中普遍存在。在本章中，将描述用碱性3,3'-二氨基联苯胺（DAB）对过氧化氢酶进行细胞化学染色的方法。由于醛固定是用DAB对过氧化氢酶进行染色的前提条件，因此在细胞化学染色方法以及随后用于光镜和电镜观察的组织处理之前，先介绍一种用戊二醛对大鼠肝脏进行灌注固定的方法。