植物细胞中微管成像的策略。

Strategies for imaging microtubules in plant cells.

作者信息

Holzinger Andreas, Kawamura Eiko, Wasteneys Geoffrey O

机构信息

Institute of Botany, Department of Physiology and Cell Physiology, University of Innsbruck, Innsbruck, Austria.

出版信息

Methods Mol Biol. 2009;586:243-62. doi: 10.1007/978-1-60761-376-3_13.

DOI:10.1007/978-1-60761-376-3_13

PMID:19768434

Abstract

Microtubules are required throughout plant development for a wide variety of processes, and different strategies have been evolved to visualize them. This chapter summarizes the most effective of these methods and points out potential problems and pitfalls. We outline the freeze-shattering method for immunolabeling microtubules in aerial organs such as leaves that require mechanical permeabilization, discuss current options for live cell imaging of MTs with fluorescently tagged proteins (FPs), and provide different fixation protocols for preserving MTs for transmission electron microscopy including chemical fixation, high pressure freezing/freeze substitution, and post-fixation staining procedures for transmission electron microscopy.

摘要

微管在植物整个发育过程中的多种进程中都是必需的，并且已经演化出了不同的策略来对其进行可视化观察。本章总结了这些方法中最有效的方法，并指出了潜在的问题和陷阱。我们概述了用于对诸如叶子等地上器官中的微管进行免疫标记的冷冻破碎法，这种方法需要机械通透处理；讨论了使用荧光标记蛋白（FPs）对微管进行活细胞成像的当前方法；并提供了不同的固定方案，用于保存微管以便进行透射电子显微镜观察，包括化学固定、高压冷冻/冷冻置换以及透射电子显微镜的后固定染色程序。