Fluorescence live cell imaging.
Fluorescence live cell imaging.
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DOI：
10.1016/b978-0-12-420138-5.00005-7
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发表时间：
2014
影响因子：
--
通讯作者：
Wittmann, Torsten
中科院分区：
文献类型：
--
作者：
Ettinger, Andreas;Wittmann, Torsten
活细胞的荧光显微镜已经成为现代细胞生物学的一个组成部分。荧光蛋白标签、活细胞染料和其他荧光标记感兴趣蛋白质的方法提供了一系列工具，可以在显微镜下研究几乎任何细胞过程。收集有意义的活细胞显微镜数据的两个主要实验挑战是在保持有用的信噪比的同时最大限度地减少光损伤，并为细胞或组织提供合适的环境来复制生理细胞动力学。本章的目的是给一个总体概述的荧光活细胞成像，适用于大多数荧光显微镜模式，并在环境控制的重点是哺乳动物组织培养细胞的显微镜设计选择的关键。此外，我们提供了指导如何设计和评估荧光蛋白结构的旋转盘共聚焦显微镜。
Fluorescence microscopy of live cells has become an integral part of modern cell biology. Fluorescent protein tags, live cell dyes, and other methods to fluorescently label proteins of interest provide a range of tools to investigate virtually any cellular process under the microscope. The two main experimental challenges in collecting meaningful live cell microscopy data are to minimize photodamage while retaining a useful signal-to-noise ratio, and to provide a suitable environment for cells or tissues to replicate physiological cell dynamics. This chapter aims to give a general overview on microscope design choices critical for fluorescence live cell imaging that apply to most fluorescence microscopy modalities, and on environmental control with a focus on mammalian tissue culture cells. In addition, we provide guidance on how to design and evaluate fluorescent protein constructs by spinning disk confocal microscopy.