ABSTRACT
The evolution of time domain spectroscopy as an analytical tool has provided a viable method for monitoring various photophysical phenomena in chemistry and biochemistry, including the structure and dynamics of proteins [1], rotational diffusion in restricted environments [2], and excited state proton transfer reactions [3]. In addition, fluorescence lifetime determinations have been used as a detection scheme in many analytical applications such as capillary electrophoresis [4], liquid chromatography [5], fluorescence microscopy [6-10], determination of adsorption modes on chromatographic stationary phases [11], and measurements of fluorescence lifetimes for single molecular events [12,13]. There are many advantages of time domain measurements compared to steadystate techniques, with the main advantage being that fluorescence measurements in the time domain yield information about the reaction rates of intra- and intermolecular processes. In addition, fluorescence lifetime measurements provide a method to probe the local environment of the fluorophore [14]. Moreover, under appropriate conditions, lifetimes can be determined with higher precision than intensity-based measurements.
