Fluorescence decays of flavin adenine dinucleotide (FAD) that is a typical autofluorescent species in cells and tissues have been measured in a mixture of alcohol and water in the femtosecond and nanosecond time range. The fluorescence lifetimes of both the stacked conformation between the isoalloxazine and adenine moieties in close proximity and the extended open conformation in water are affected by the addition of alcohol. The nanosecond fluorescence lifetime of the open conformation increases with decreasing dielectric constant of the medium, contributing to the enhancement of the fluorescence intensity of FAD in less dielectric media. The fluorescence lifetime of the open conformation is also affected by medium viscosity, suggesting that the photoexcited open conformation is quenched by the dynamic interaction between the two aromatic rings. The fluorescence component decaying in tens of picoseconds is attributed to the stacked conformation that shows the efficient fluorescence quenching due to the intramolecular electron transfer. The picosecond fluorescence lifetime of the stacked conformation increases with decreasing dielectric constant, suggesting the shift of the distribution of the stacked conformation to a longer intramolecular distance between the two aromatic rings in less dielectric media. The pre-exponential factor of the picosecond decaying component relative to that of the nanosecond one decreases with the increase of the alcohol concentration in the femtosecond time-resolved fluorescence, which demonstrates the increase in the population of the open conformation with the reduction of the dielectric constant. The possibility to evaluate the polar environment in a cell by the fluorescence lifetime of FAD is discussed based on the results obtained.
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