FRET Microscopy for Real-Time Visualization of Second Messengers in Living Cells
FRET Microscopy for Real-Time Visualization of Second Messengers in Living Cells
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DOI：
10.1007/978-1-4939-6810-7_6
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发表时间：
2017-01-01
期刊：
影响因子：
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通讯作者：
Nikolaev, Viacheslav O.
中科院分区：
文献类型：
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作者：
Kraft, Axel E.;Nikolaev, Viacheslav O.
福斯特共振能量转移（FRET）显微镜是分子生物学和医学研究中的有用工具，用于监测和量化蛋白质-蛋白质相互作用和生物化学过程的实时动态。使用这种成熟的技术，可以在完整的细胞或组织，甚至在各种亚细胞区室中研究许多新的信号传导机制。在这里，我们描述了如何在表达FRET生物传感器的活细胞中进行FRET测量，以及如何评估这些数据。该通用协议可应用于各种荧光生物传感器的FRET测量。
Forster Resonance Energy Transfer (FRET) microscopy is a useful tool in molecular biology and medical research to monitor and quantify real-time dynamics of protein-protein interactions and biochemical processes. Using this well-established technique, many novel signaling mechanisms can be investigated in intact cells or tissues and even in various subcellular compartments. Here, we describe how to perform FRET measurements in living cells expressing FRET-based biosensors and how to evaluate these data. This general protocol can be applied for FRET measurements with various fluorescent biosensors.