费斯特共振能量转移
生物
共焦
生物物理学
单分子微动
共焦显微镜
分辨率(逻辑)
蛋白质-蛋白质相互作用
计算生物学
生物系统
细胞生物学
荧光
物理
计算机科学
光学
人工智能
作者
Ammasi Periasamy,Horst Wallrabe,Ye Chen,Margarida Barroso
标识
DOI:10.1016/s0091-679x(08)00622-5
摘要
Förster resonance energy transfer (FRET) is an effective and high resolution method to monitor protein-protein interactions in live or fixed specimens. FRET can be used to estimate the distance between interacting protein molecules in vivo or in vitro using laser-scanning confocal FRET microscopy. The spectral overlap of donor and acceptor-essential for FRET-also generates a contamination of the FRET signal, which should be removed in order to carry out quantitative data analysis with confidence. Quantitative FRET data analysis addresses the wealth of information contained in the data set, once optimized FRET imaging has been completed. In this chapter, we describe step-by-step what the issues are in quantitative FRET data analysis, using membrane receptor trafficking and organization as an example. The assays described are applicable to many other biological applications.
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