Imaging Protein-Protein Interactions by Förster Resonance Energy Transfer (FRET) Microscopy in Live Cells

Q1 Biochemistry, Genetics and Molecular Biology

Current Protocols in Protein Science Pub Date : 2018-07-09 DOI:10.1002/cpps.58

Javier Manzella-Lapeira, Joseph A. Brzostowski

引用次数: 7

Abstract

This updated unit compares three methods to acquire Förster Resonance Energy Transfer (FRET) data in living cells using a confocal microscope: Acceptor photobleaching, Acceptor-sensitized emission FRET, and Donor fluorescence lifetime imaging. Detailed protocols for live cell husbandry, image acquisition, and data analysis are provided. In addition to providing instructions for manufacturer's analysis tool sets, we provide an easy-to-use, MATLAB-based code to calculate FRET efficiency from data obtained using the Acceptor photobleaching or Acceptor-sensitized emission method, which can be freely downloaded. © 2018 by John Wiley & Sons, Inc.

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通过Förster共振能量转移(FRET)显微镜在活细胞中成像蛋白质-蛋白质相互作用

这个更新的单元比较了三种方法获得Förster共振能量转移(FRET)数据在活细胞使用共聚焦显微镜:受体光漂白，受体敏化发射FRET，和供体荧光寿命成像。提供了活细胞饲养、图像采集和数据分析的详细协议。除了为制造商的分析工具集提供说明外，我们还提供了一个易于使用的基于matlab的代码，可以根据使用Acceptor光漂白或Acceptor敏化发射方法获得的数据计算FRET效率，该代码可以免费下载。©2018 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Current Protocols in Protein Science Biochemistry, Genetics and Molecular Biology-Biochemistry

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期刊介绍： With the mapping of the human genome, more and more researchers are exploring protein structures and functions in living organisms. Current Protocols in Protein Science provides protein scientists, biochemists, molecular biologists, geneticists, and others with the first comprehensive suite of protocols for this growing field.