An endoplasmic reticulum-specific ratiometric fluorescent probe for imaging esterase in living cells

Esterase is primarily distributed in the endoplasmic reticulum (ER) and often overexpressed in cancer cells. Therefore, the detection of esterase in ER is significant for monitoring the metabolic process of various esters and evaluating the efficacy of chemotherapeutic prodrugs. However, only few fluorescent probes can detect esterase in the ER due to the lack of ER-specificity. More seriously, these probes are often limited by low pearson's colocalization coefficient and one single wavelength emission. To solve those problems, an ER-specific ratiometric fluorescent probe (ER-EST) is designed for detecting esterase in living cells. The ER-EST shows a ratiometric and red-shifted emission (125 nm) from 435 to 560 nm after hydrolysis by esterase. The fluorescence intensity ratio of ER-EST displays quantitative response to the esterase activity (0–0.5 U/mL) with low detection limit of 1.8 × 10^-4 U/mL. Importantly, the ER-EST with good biocompatibility and excellent ER-targeted ability was successfully employed to ratiometric image the endogenous endoplasmic reticulum esterase in living cells.