Versatile Ratiometric Fluorescent Probe Based on the Two-Isophorone Fluorophore for Sensing Nitroxyl

Nitroxyl (HNO) is closely linked with numerous biological processes. Fluorescent probes provide a visual tool for determining HNO. Due to fluorescence quenching by HNO-responsive recognition groups, most of the current fluorescent probes exhibit an "off-on" fluorescence response. As such, the single fluorescence signal of these probes is easily affected by external factors such as the microenvironment, sensor concentration, and photobleaching. Herein, we have developed a ratiometric fluorescent probe (CHT-P) based on our previously developed two-isophorone fluorophore. CHT-P could be used to determine HNO through ratiometric signal readouts with high selectivity and sensitivity, ensuring the accurate quantitative detection of HNO. Additionally, the probe exhibited low cytotoxicity, was cell permeable, and could be used for ratiometric imaging of HNO in cells. Finally, CHT-P-coated portable test strips were used to determine HNO using the solid-state fluorescence signal readout.