A broad-spectrum fluorescent probe, which can be applied to monitoring H₂S in various biological systems, has been rationally designed and synthesized. This specific probe was applied to localize the endogenous H₂S in living Raw264.7 macrophage cells, HepG2 cells, and H9C2 cells. At the same time, the probe has successfully visualized CBS- and CSE- induced endogenous H₂S production and monitored CBS and CSE activity in H9C2 cells. This probe could serve as a powerful molecular imaging tool to further explore the physiological function and the molecular mechanisms of endogenous H₂S in living animal systems.