A novel dicyanoisophorone (DCI)-based NIR fluorophore employing 2, 4-thiazolidinediones as the modification site was designed for fluorescence imaging. The fluorophore was assessed as a switchable reporter for H2O2 and the probe exhibited lysosomes-targeted, a large turn-on fluorescence signal at 720 nm with a large stokes shift (150 nm) and can be used in biological systems. The ability of the novel fluorophore to emit NIR fluorescence through a “turn-on” activation mechanism makes it a promising fluorophore for in vivo imaging applications. The strategy of introducing the thiazolidinediones with the easy modification site into the fluorophore has a good application prospect to expand the application of the NIR fluorophore.