Ultrasensitive and specific two-photon fluorescence detection of hypochlorous acid by a lysosome-targeting fluorescent probe for cell imaging

Hypochlorous acid (HOCl) is involved in numerous cellular processes, such as pathogen response, immune regulation, and anti-inflammation. Consequently, the development of HOCl detection at the cellular level has been an important issue in investigating the dynamic distributions of HOCl. Herein, a fluorescent probe, Lyso-NA, containing a HOCl-reactive aminophenol group and a lysosomal-targeting morpholine group, has been effectively designed for detecting lysosomal HOCl. The reaction of Lyso-NA with HOCl induces the oxidation of aminophenol and accompanied by a 136-fold fluorescence enhancement. The detection limit is found at 13 nM. The fluorescence enhancement is accomplished through the suppression of twisted intramolecular charge transfer (TICT). With morpholine, the probe Lyso-NA shows the great lysosomal targetable ability for imaging endogenous lysosomal HOCl in living cells and tissues by two-photon microscopy, providing an opportunity to monitor HOCl in the lysosomes for understanding its biological functions.