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Modem biophysical chemistry has been significantly advanced recently with rapid developments of various physical techniques and molecular biology methods.Complex biological machinery can be investigated at the most fundamental level and thus molecular mechanislms are elucidated.Here,we present our recent studies of several important biological systems by integrating femtosecond spectroscopy and site-directed mutations.We are able to map out the entire evolution of biological dynamics in real time and reveal their molecular mechanisms with unprecedented details.