Shibata Shinsuke, Murota Yukiko, Nishimoto Yoshinori, Yoshimura Mana, Nagai Toshihiro, Okano Hideyuki, Siomi Mikiko C
Department of Physiology, School of Medicine, Keio University, Shinanomachi 35, Shinjuku-ku, Tokyo, 160-8582, Japan,
Methods Mol Biol. 2015;1328:163-78. doi: 10.1007/978-1-4939-2851-4_12.
Immuno-electron microscopy and electron microscopic in situ hybridization are powerful tools to identify the precise subcellular localization of specific proteins and RNAs at the ultramicroscopic level. Here we describe detailed procedures for how to detect the precise location of a specific target labeled with both fluorescence and gold particles. Although they have been developed for the analysis of Drosophila ovarian somatic cells, these techniques are suitable for a wide range of biological applications including human, primate, and rodent analysis.
免疫电子显微镜和电子显微镜原位杂交是在超微水平上鉴定特定蛋白质和RNA精确亚细胞定位的强大工具。在这里,我们描述了如何检测同时用荧光和金颗粒标记的特定靶标的精确位置的详细程序。尽管这些技术是为分析果蝇卵巢体细胞而开发的,但它们适用于广泛的生物学应用,包括人类、灵长类和啮齿类动物分析。