A curcuminoid difluoroboron-based tumor-targeting GGT-activatable fluorescent probe (Glu-DFB-biotin) is designed, which has remarkable characteristics of dual-channel and light-up near-infrared (NIR) fluorescence. Furthermore, Glu-DFB-biotin can differentiate hepatic cancer cells from normal cells and is successfully employed for real-time tracking of endogenous GGT activity in living cells.