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Abstract

Real-time imaging and quantification of adenosine triphosphate (ATP) dynamics in living cells are important for understanding relationship between energy metabolism and cell functions. However, few synthetic fluorescent probes have been reported to tackle this challenge due to lack of accurate fluorescence readout and suitable response concentration. In this work, we developed a ratiometric fluorescent probe (Rh6G-ACFPN) for quantitatively monitoring the fluctuation of mitochondrial ATP in living cells. Rh6G-ACFPN selectively and reversibly responds to ATP with an ideal dissociation constant (Kd) of 4.72 mM (3–10 mM: the range of mitochondrial ATP concentrations). Live-cell imaging allows us to directly monitor the dynamic changes of mitochondrial ATP in high temporal resolution. Moreover, for the first time, mitochondrial ATP in normal and cancer cells lines was successfully quantified and discriminated. These results demonstrate the versatility of Rh6G-ACFPN as a useful imaging tool to elucidate the function of mitochondrial ATP in living cells.