A fluorescent probe with dual acrylate sites for discrimination of different concentration ranges of cysteine in living cells

Highlights: • A novel fluorescent probe with dual acrylate sites was designed and synthesized. • The probe enables discrimination of different concentration ranges of cysteine. • Same reacting groups in one probe with different steric hindrances could exhibit different reactivities to cysteine. • This probe was successfully applied for monitoring of cysteine concentration in living cells. Monitoring of cysteine (Cys) is of significant importance for studying Cys-involved biological functions and clinically diagnosing Cys-related diseases. Recently, few fluorescent probes with two different reacting sites were reported to be capable of sensing different concentration ranges of Cys with distinct fluorescence signals, particularly suiting for bioimaging. However, due to relative sophisticated synthesis and moderate selectivity, the applications of these probes were still severely restricted. In this work, we proposed a novel probe design strategy by utilizing two same reacting groups, instead of two different reacting groups, to simplify the synthesis route and minimize the interference from competing species. Same reacting groups in a probe with different steric hindrances could exhibit different reactivities to Cys. This probe showed distinguishable fluorescence peak wavelengths towards low and high concentration ranges of Cys, giving green and blue emissions, respectively. Moreover, this probe was successfully applied for monitoring of Cys concentration in living cells. We believe this work provided a simpler strategy for dual-site fluorescent probes to sense difference concentration ranges of Cys, which may inspire more probe design in future.