Mitochondria-Targeted Near-Infrared Fluorescent Off-On Probe for Selective Detection of Cysteine in Living Cells and in Vivo.

Cysteine (Cys) plays crucial roles in biological systems and in mitochondrial processes. Highly selective probes for specific detection of mitochondrial Cys over other biological thiols are rare. Herein, we designed and synthesized a mitochondria-targetable near-infrared (NIR) fluorescent off-on probe, NFL1, based on a fluorescein derivative for Cys detection. Probe NFL1 has a lipophilic cation unit as the mitochondria biomarker and an acrylate group as the Cys-recognition unit as well as a fluorescence quencher. The probe itself is nonfluorescent due to the photoinduced electron transfer process. Upon addition of Cys, marked enhancement in the NIR emission (735 nm) can be monitored due to cleavage of the acrylate moiety. This probe had great sensitivity and selectivity for the rapid detection of Cys over homocysteine (Hcy) and glutathione (GSH) with an ultralow detection limit of 14.5 nM. More importantly, the probe successfully targeted mitochondria, detected endogenous Cys, and assessed mitochondrial oxidative stress in living cells. Probe NFL1 was also capable of detecting and imaging Cys in living nude mice, indicating its significant potential in biological applications.