Laser instrumentation and data analysis in time correlated photon counting fluorescence decay measurements in biochemical investigations

摘要

In the past several years the utility of fluorescence decay measurements in biological and biochemical studies has been clearly demonstrated. The time correlated single photon counting (TCSPC) technique has been used extensively in these experiments. Improvements in the instrumentation and data analysis have allowed higher sensitivities with a higher temporal and spectral resolution of the data. The results have led to new insights into the understanding of conformational heterogeneity and dynamic processes in biochemical systems. The combination of picosecond high repetition rate pulsed laser excitation with microchannel plate photomultiplier detectors has been a major advance in the instrumentation. Concomitant with these instrumental advances data analysis methods have also made great progress. These developments are reviewed and recent examples of applications to studies of biomolecules are presented.