Weidemann Thomas
BIOTEC, Biophysics, Technische Universität Dresden, Dresden, Germany.
Methods Mol Biol. 2014;1076:539-55. doi: 10.1007/978-1-62703-649-8_24.
Fluorescence correlation spectroscopy (FCS) can add dynamic molecular information to images of live cells. For example, a confocal laser scanning microscope (CLSM) equipped with an accessory FCS unit provides the possibility to first image the spatial distribution of a fluorescent protein before probing its mobility within defined regions of interest. Whereas specific protein-protein interactions are preferably assayed with a dual-color approach, single-color FCS can still provide valuable information about the size of the diffusing entities and potential interactions with other, nonfluorescent, proteins or subcellular structures. Because number fluctuations are measured, the concentrations of freely diffusing complexes and their state of oligomerization are accessible.
荧光相关光谱法（FCS）能够为活细胞图像增添动态分子信息。例如，配备了辅助FCS单元的共聚焦激光扫描显微镜（CLSM）使得在探测荧光蛋白在特定感兴趣区域内的流动性之前，先对其空间分布进行成像成为可能。虽然特定的蛋白质-蛋白质相互作用最好采用双色方法进行检测，但单色FCS仍可提供有关扩散实体大小以及与其他非荧光蛋白质或亚细胞结构潜在相互作用的有价值信息。由于测量的是数量波动，因此可以获取自由扩散复合物的浓度及其寡聚化状态。
