The mitochondrial targeting and viscosity-sensitive near-infrared fluorescence probe TQS-OH was an aggregation induced emission luminogen constructed by regulating the receptor structure and hydroxylating its terminal. TQS-OH presented more than 27.35-fold fluorescence enhancement at the maximum emission peak when the viscosity level increased from 0.49 to 1048 cP. It also appeared excellent reactive oxygen species production ability under white light irradiation and appeared strong photodynamic therapied effect on HepG2 cells. In addition, cellular co-location analysis implied that TQS-OH had accurate mitochondrial targeting ability, with co-location coefficient up to 0.95 in human hepatocellular carcinoma HepG2 cells and it also successfully achieved fatty liver imaging. Meanwhile, we found that TQS-OH induced cancer cell apoptosis by activating mitochondrial apoptosis pathway. Furthermore, TQS-OH ultimately has implemented the tumor photodynamic therapy with high efficiency in vivo, with a tumor inhibition rate as high as 50.77%. The above results made it a promising aggregation induced emission material for the diagnosis of fatty liver and tumor phototherapy.