Cmarko Dusan, Koberna Karel
Institute of Cellular Biology and Pathology, Ist Faculty of Medicine, Charles University, Prague, Czech Republic.
Methods Mol Biol. 2007;369:213-28. doi: 10.1007/978-1-59745-294-6_11.
Electron microscopy in situ hybridization (EM-ISH) represents a powerful method that enables the localization of specific sequences of nucleic acids at high resolution. We provide here an overview of three different nonisotopic EM-ISH approaches that allow the visualization of nucleic acid sequences in cells. A comparison of various methods with respect to their sensitivity and the structural preservation of the sample is presented, with the aim of helping the reader to choose a convenient hybridization procedure. The post-embedding EM-ISH protocol that currently represents the most widely used technique is described in detail, with a special emphasis on the organization of the cell nucleus.
电子显微镜原位杂交（EM-ISH）是一种强大的方法，能够在高分辨率下定位特定的核酸序列。我们在此概述三种不同的非同位素EM-ISH方法，这些方法可使细胞中的核酸序列可视化。本文还比较了各种方法在灵敏度和样品结构保存方面的差异，旨在帮助读者选择合适的杂交程序。文中详细描述了目前应用最广泛的包埋后EM-ISH方案，并特别强调了细胞核的结构。
