Abstract
Since its invention by Magde, Elson, and Webb (1) in the seventies of the last century, fluorescence correlation spectroscopy (FCS) has become a widely used and powerful technique in biophysical research (2). It is most often used for studying the mobility of molecules (diffusion, directed transport) (3) but also for looking at the kinetics of photophysical transitions (4,5), the temporal dynamics of conformation fluctuations (6), or the speed of rotational diffusion (7,8). The core idea is to record long time traces of the fluorescence intensity that is excited in and detected from the picoliter-sized detection volume of a confocal microscope and then to calculate the temporal correlation function of the observed intensity fluctuations.
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