The viscosity of the cell microenvironment is a parameter that affects cell physiological processes. A fluorescent probe X-V was designed to detect the viscosity changes of a hepatic ischemia-reperfusion injury (HIRI) cell model with high selectivity and sensitivity. The fluorescence emission wavelength is 615 nm and the Stokes shift can be up to 125 nm, which can be used not only for intracellular viscosity changes stimulated by different drugs but also for the detection of cell viscosity changes in the HIRI cell model. Probe X-V provides a useful tool to study the relationship between mitochondrial viscosity and related diseases.