Ultrasensitive fluorescent probe for visual biosensing of esterase activity in living cells and its imaging application.

Esterase activity is often used as an index to evaluate the health status of cells and plays an important role in cell metabolism and apoptosis. Herein, we develop two fluorescent probes for visual biosensing of esterase activity and imaging in living cells. In vitro, after the introduction of esterase, enzymolysis destroys the ester bond of the probe, causing the fluorescent color of probe changes from yellow to red, thus realizing the visual strategy for determination of esterase activity, with high sensitivity and selectivity. Especially, probe VA, 2-(4-acetoxystyryl)-3-ethyl-1,1-dimethyl- 1H-benzo[e]indol-3-ium, exhibits higher sensitivity with a lower detection limit (up to 7.15 × 10 U/mL). In the cell experiment, the fluorescent probe VA also shows good biocompatibility and high spatial resolution, and is successfully applied to the intracellular fluorescent imaging and biosensing of esterase in living cells. More importantly, the probe VA can judge the unhealthy state of HO-induced HeLa cells using dual-fluorescence signals. The results confirm that the fluorescence method is a reliable tool for detecting endogenous esterase in living biological system.