Elaborating the fluorescence regulation and quenching mechanism of sulfur‐for‐oxygen replacement for fluorophores

Abstract Thio‐caged fluorophores can be effectively desulfurized into their oxygenated derivatives through visible light, thereby restoring the strong emission of fluorophores, and are applied in the field of live cell super‐resolution imaging. Herein, we theoretically investigated the reasons for the low fluorescence quantum yields of a series of thio‐caged fluorophores and the underlying reasons for the differences in fluorescence quantum yields of their oxygenated derivatives. The calculation results show that the S atom on the thiocarbonyl group is more likely to excite n electrons to form the nπ* state, which reduces the energy of the nπ* state and leads to fluorescence quenching. In contrast, the O atom on the carbonyl group is more likely to excite π electrons to form ππ* state, which is the main reason for restoring the strong emission of fluorophore. Meanwhile, the calculation results show that the difference of fluorescence intensity caused by oxygenated derivatives is determined by the number of the carbonyl group, which affects the vibronic coupling between ππ* and nπ* states and thereby leads to fluorescence quenching. These results can effectively reveal the fluorescence quenching mechanism of thio‐caged fluorophores and the luminescence mechanism of their oxygenated derivatives, and provide correct and guiding design strategies for the development of new thio‐caged fluorophores.