通过铽柠檬酸蒸气在电子显微镜下进行特定 RNA 可视化。

Specific RNA Visualization at Electron Microscopy via Terbium Citrate Vapors.

机构信息

Department of Biology and Biotechnology "Lazzaro Spallanzani", Laboratory of Cell Biology and Neurobiology, University of Pavia, Pavia, Italy.

出版信息

Methods Mol Biol. 2023;2566:233-240. doi: 10.1007/978-1-0716-2675-7_19.

DOI:10.1007/978-1-0716-2675-7_19

PMID:36152256

Abstract

Terbium citrate staining represents the method of choice for RNA visualization at transmission electron microscopy. Because of its affinity for guanosines in RNA rather than in DNA, terbium citrate gives precise results being a selective staining. However, difficulties often arise when performing this technique, especially in crucial and delicate steps such as rinses, when it is not uncommon to excessively reduce the already feeble contrast. For these reasons, we developed a straightforward and secure approach to overcome such complications. Here we report a new method for RNA single molecule localization by means of terbium citrate vapors, viable for every type of fixation and embedding.

摘要

柠檬酸铽染色是在透射电子显微镜下观察 RNA 的首选方法。由于其与 RNA 中的鸟嘌呤而非 DNA 中的鸟嘌呤具有亲和力，因此柠檬酸铽是一种选择性染色，结果非常准确。然而，在进行这项技术时，尤其是在冲洗等关键和精细的步骤中，往往会出现困难，因为已经很微弱的对比度很容易被过度降低。出于这些原因，我们开发了一种简单而安全的方法来克服这些并发症。在这里，我们报告了一种通过柠檬酸铽蒸气对 RNA 单分子进行定位的新方法，适用于各种固定和包埋方式。