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Hydrogen sulfide (H 2 S) is a vital gas signaling molecule that performs crucial physiological functions in biological systems, and it can also cause food safety problems. In this study, we developed a cyanine derivative-based mitochondrial-targeted near-infrared fluorescent probe PC-H 2 S for detecting H 2 S. When PC-H 2 S reacted with H 2 S, it reached maximum fluorescence intensity at 656 nm and detection limit as low as 34 nM, and exhibited strong chromogenic response from green to red for H 2 S. Moreover, PC-H 2 S could be utilized to identify H 2 S generated during food spoilage and thus monitored the process of food spoilage. More significantly, PC-H 2 S showed excellent mitochondrial targeting capabilities and endogenous/exogenous H 2 S imaging of living cells and zebrafish could be performed. As a result, this research provides an attractive method for detecting H 2 S in food and biological systems and has potential value in physiopathological research. [Display omitted] • A cyanine derivative-based mitochondrial-targeted NIR fluorescent probe PC-H 2 S for detecting H 2 S was developed. • PC-H 2 S had a low detection limit and exhibited strong chromogenic response from green to red for H 2 S in naked eyes and UV light. • PC-H 2 S test strips was capable of monitoring H 2 S produced by food spoilage. • PC-H 2 S could be used for living cells and zebrafish imaging. [ABSTRACT FROM AUTHOR]