The ratiometric fluorescent probes for monitoring the reactive inorganic sulfur species (RISS) signal in the living cell

RSSs (reactive sulfur species) and their metabolites, such as H2S, Sn2−, SO32−/HSO3−, S2O42− and S2O52− (Reactive Inorganic Sulfur Species, RISSs), play a crucial role in the cushion against oxidative stress and the other physiological events. The molecular mechanisms how they affect cellular signaling and other physiological events remain largely unknown. To address their physiological functions, the techniques that can track their levels should be invaluable. Herein, six coumarin hemicyanine scaffolds (CH-RISSs) were synthesized and their fast and strong responses upon H2S, Sn2−, SO32−, HSO3−, S2O42− and S2O52− (Reactive Inorganic Sulfur Species, RISSs) were clarified in the absorption (colorimetric) and fluorescence (ratiometric) spectra, which showed good stability in the physiological pH (7.4). Upon the analytes, the maxima absorption of CH-RISSs switched from ~580 nm to ~400 nm in the absorption spectra. The fluorescence of CH-RISSs depleted at 650–660 nm and increased at 480–505 nm upon the RISSs. Both of coumarin hemicyanine structures with C12 alkyl chain (CH-RISS-3 and CH-RISS-6) showed quick and robust ratiometric fluorescence switch in the living cell imaging. Access to the fluorescent probes for RISSs sets the stage for applying the developing technologies to probe reactive sulfur biology in living systems.