Experimental Study for the Effect of Photobleaching on Reaction Rate Constant

Abstract

Fluorescence is a useful phenomenon to detect the biomolecule and the chemical reaction. Fluorescence based detection techniques have been developed with microscope of good performance and super resolution microscopy which is able to even detect each single molecule. But there is still a remaining problem to obtain the kinetics using fluorescence in single-molecule level. The problem is about photobleaching. Photobleaching is photochemical destruction that makes dye not to emit fluorescence. And photobleaching interrupts to detect the signal from fluorescence of chemical reaction from the beginning to the end of the reaction. Furthermore, the effect on growing down the detectable fluorophore, photobleaching is one of the main obstacles to gain the reaction rate. There were attempts to avoid photobleaching, but the limitation of specimen or techniques it is hard to study in single-molecule level. In this study, the way to overcome photobleaching of fluorescent dye and its examples will be introduced. The problem of studying single-molecule with fluorescence in single-molecule level will be presented and its solution too. In short, the effects of photobleaching on single-molecule experiments and theory to overcome the effects of photobleaching with experimental examples will be introduced.