Fluorescence imaging holds great potential as a powerful diagnostic tool for tumor cell visualization. However, a significant challenge in fluorescence imaging is the high background signal, which obscures the tumor-specific signal and reduced the signal-to-noise ratio (SNR) of imaging, thereby reducing the accuracy of tumor detection and delineation. In this research, we designed and synthesized an amphiphilic disulfide dicyanine ss-diCy7, which can self-assemble into nanoparticles with uniform dispersion in aqueous environments. The fluorescence intensity of these nanoparticles is significantly reduced by 96% due to aggregation-induced quenching arising from π-π stacking. The nanoparticles exhibit a highly specific response to glutathione (GSH) in vitro, resulting in a substantial enhancement of fluorescence intensity by a 24-fold. The enhancement was also achieved in cell and mouse imaging experiments. In addition, in the mouse tumor model, ss-diCy7 nanoparticles demonstrated superior performance compared to traditional mono-cyanine dyes, offering a reduced background signal and prolonged fluorescence duration. This work is anticipated to contribute to the high-resolution tumor imaging.