Mitochondria-Anchored Colorimetric and Ratiometric Fluorescent Chemosensor for Visualizing Cysteine/Homocysteine in Living Cells and Daphnia magna Model.

Cysteine (Cys) and homocysteine (Hcy) are essential for maintaining the cellular redox homeostasis and play critical roles in pathological and physiological processes. The development of Cys/Hcy-specific responsive fluorescent probes that are independent of the surrounding environment, equipment, and abundant endogenous GSH is critical to accurately investigate the roles of Cys/Hcy in living biological systems. In this work, a novel ratiometric and mitochondria-anchored fluorescence chemosensor, PYR, was constructed on the basis of 4-methylphenol-substituted pyronin fluorophore. The probe exhibited ratiometric fluorescence emission (F540 nm/F620 nm) for the detection of Cys/Hcy with high selectivity, sensitivity (Cys, 22 nM; Hcy, 23 nM), rapid response (Cys, 5 min), and a merit enhancement of ratio fluorescent signal (Cys, 163-fold; Hcy, 125-fold). The probe showed excellent membrane permeability and was applied to visualize mitochondrial biothiols in living cells under H2O2-induced redox imbalance, kidney tissues with a penetration depth of 100 μm, and Daphnia magna model for the first time. The results demonstrate that PYR will provide a promising platform for the diagnosis of thiol-related diseases.