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用于电子显微镜下分子亚细胞检测的预包埋标记

Pre-embedding labeling for subcellular detection of molecules with electron microscopy.

作者信息

Polishchuk Elena V, Polishchuk Roman S

机构信息

Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, Pozzuoli, NA, 80078, Italy; ITMO University, Kronverkskiy pr., 49, St. Petersburg, 197101, Russia.

Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, Pozzuoli, NA, 80078, Italy.

出版信息

Tissue Cell. 2019 Apr;57:103-110. doi: 10.1016/j.tice.2018.11.002. Epub 2018 Nov 20.

Abstract

Labeling of specific molecules for electron microscopy provides an extremely powerful tool to investigate subcellular distribution of proteins, lipids and sugars with resolution of a few nanometers. Moreover, this technology offers a unique opportunity to see how the pattern of these molecules changes in different physiological or pathological conditions. Pre-embedding labeling evolved as one of the easiest method for molecule detection in electron microscopy. It was named "pre-embedding" because the procedure involves a labeling step before the embedding of the specimen in the resin, its subsequent sectioning and analysis of sections under the electron microscope. Here we review different strategies and technical tips of the pre-embedding procedure, the potential of this method for detection and quantification of molecular components at the ultrastructural level, and the integration of the pre-embedding approach with rapidly developing light and electron microscopy technologies.

摘要

用于电子显微镜的特定分子标记提供了一种极其强大的工具,可用于研究蛋白质、脂质和糖类的亚细胞分布,分辨率可达几纳米。此外,这项技术提供了一个独特的机会,来观察这些分子的模式在不同生理或病理条件下是如何变化的。预包埋标记作为电子显微镜中检测分子最简单的方法之一而发展起来。它被称为“预包埋”,是因为该过程包括在将标本包埋在树脂中之前的标记步骤、随后的切片以及在电子显微镜下对切片进行分析。在这里,我们回顾预包埋过程的不同策略和技术要点、该方法在超微结构水平检测和定量分子成分的潜力,以及预包埋方法与快速发展的光学和电子显微镜技术的整合。

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