Wouters F S, Bastiaens P I
Imperial Cancer Research Fund, London, United Kingdom.
Curr Protoc Cell Biol. 2001 May;Chapter 17:Unit 17.1. doi: 10.1002/0471143030.cb1701s07.
FRET microscopy enables the detection of different biochemical states of proteins in cells. The use of fluorescence in the detection of proteins, by chemical modification, by immunofluorescence, or by genetic encoding of a green fluorescent protein fusion protein, provides more information than just the location of the protein in the cell. The properties of the fluorophore can be exploited to extract information on protein-protein interactions. A straightforward, quantitative imaging approach is presented to measure FRET that is based on internal calibration by acceptor photobleaching.
荧光共振能量转移显微镜可检测细胞中蛋白质的不同生化状态。通过化学修饰、免疫荧光或绿色荧光蛋白融合蛋白的基因编码，利用荧光检测蛋白质，所提供的信息不仅仅是蛋白质在细胞中的位置。可以利用荧光团的特性来提取有关蛋白质-蛋白质相互作用的信息。本文提出了一种基于受体光漂白进行内部校准来测量荧光共振能量转移的直接定量成像方法。
