A lysosome-targetable fluorescent probe for real-time imaging cysteine under oxidative stress in living cells

As an effective lysosomal biomarker for oxidative stress status, cysteine (Cys) plays an important role in lysosomal proteolysis. Herein, we report the first lysosome-targetable fluorescence probe (MCAB) for Cys-selective detection based on nucleophilic addition reaction of sulfhydryl toward a α, β-unsaturated ketone and demonstrate its application to lysosomal-targetable imaging. MCAB is designed based on a α, β-unsaturated ethanoylcarbazole as the fluorophore and the thiols reaction site, and a methylcarbitol unit as a lysosome-targetable group. Upon reacting with Cys, this probe turns on highly specific fluorescence signals linearly proportional to Cys concentrations over the range of 0-300 μM. MCAB detects Cys with a rapid response time (within 12 min) and low limit of detection (0.38 μM). MCAB is highly selective to Cys over other similar biothiols including homocysteine (Hcy) and glutathione (GSH). Moreover, it also exhibits significant lysosomal-targetable ability, which is ideal for lysosomal Cys-selective imaging. Using MCAB, we have successfully visualized the fluctuation endogenous Cys in lysosomes under oxidative stress status in real-time.