FRET microscopy in the living cell: Different approaches, strengths and weaknesses
FRET microscopy in the living cell: Different approaches, strengths and weaknesses
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DOI：
10.1002/bies.201100086
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发表时间：
2012-05-01
期刊：
影响因子：
4
通讯作者：
Tramier, Marc
中科院分区：
文献类型：
--
作者：
Padilla-Parra, Sergi;Tramier, Marc
定量荧光显微镜中的新成像方法，例如福斯特共振能量转移（FRET），在过去几年中得到了发展，并开始广泛应用于生物学问题。 FRET 用于检测和定量蛋白质相互作用和生化活性。在此，我们回顾了显微镜中测量 FRET 的不同方法，更重要的是，它们的优点和缺点。我们认为，荧光寿命成像显微镜（FLIM）有利于定量检测分子间相互作用，强度比方法代表了检测分子内 FRET 的有效且直接的选择。用于定量和快速时空蛋白质相互作用研究的单分子技术和数据分析的有前途的方法为生物学研究中的 FRET 开辟了新的途径。
New imaging methodologies in quantitative fluorescence microscopy, such as Forster resonance energy transfer (FRET), have been developed in the last few years and are beginning to be extensively applied to biological problems. FRET is employed for the detection and quantification of protein interactions, and of biochemical activities. Herein, we review the different methods to measure FRET in microscopy, and more importantly, their strengths and weaknesses. In our opinion, fluorescence lifetime imaging microscopy (FLIM) is advantageous for detecting inter-molecular interactions quantitatively, the intensity ratio approach representing a valid and straightforward option for detecting intra-molecular FRET. Promising approaches in single molecule techniques and data analysis for quantitative and fast spatio-temporal protein-protein interaction studies open new avenues for FRET in biological research.