A fluorescent light-up probe based on AIE and ESIPT processes for β-galactosidase activity detection and visualization in living cells

A novel fluorescent probe SA-βGal is reported here with light-up response to β-galactosidase. SA-βGal possesses the β-galactopyranoside group to react with β-galactosidase and releases the fluorescent salicylaldehyde azine with both aggregation induced emission (AIE) and excited-state intramolecular proton transfer (ESIPT) characteristics. The linear fluorescent response enables the in vitro quantification of β-galactosidase activity in a range of 0-0.1 U mL^-1 with a detection limit of 0.014 U mL^-1. The probe exhibits significant advantages, such as no self-quenching at high concentrations, a large Stokes shift (190 nm) and high specificity to β-galactosidase with an excellent light-up ratio of 820 fold. Moreover, thanks to its good retention in living cells, the application of SA-βGal for the imaging of cellular β-galactosidase was also achieved with high contrast.