用于电子显微镜中DNA染色的锇氨络合物

Osmium Ammine for Staining DNA in Electron Microscopy.

作者信息

Masiello Irene, Biggiogera Marco

机构信息

Department of Biology and Biotechnology, Laboratory of Cell Biology and Neurobiology, University of Pavia, Via Ferrata 9, 27100, Pavia, Italy.

出版信息

Methods Mol Biol. 2017;1560:261-267. doi: 10.1007/978-1-4939-6788-9_19.

DOI:10.1007/978-1-4939-6788-9_19

PMID:28155160

Abstract

The osmium ammine staining allows the specific detection of DNA in the cell nucleus and represents one of the most used techniques for EM cytochemistry.The procedure is a Feulgen-type reaction, consisting of an acid hydrolysis to obtain free aldehyde groups on DNA followed by their binding to osmium ammine, a Schiff-type reagent. Osmium ammine is polyamminic electron-dense compound commercially available.Here, we describe the staining procedure for ultrathin sections and the different procedures for the preparation of the reagent for acrylic and epoxy sections.

摘要

锇氨染色法可特异性检测细胞核中的DNA，是用于电子显微镜细胞化学的最常用技术之一。该程序是一种福尔根型反应，包括酸水解以在DNA上获得游离醛基，然后将其与作为席夫试剂的锇氨结合。锇氨是一种市售的多胺电子致密化合物。在此，我们描述了超薄切片的染色程序以及用于丙烯酸和环氧树脂切片的试剂制备的不同程序。