Simultaneous Detection of Hypochlorite and Singlet Oxygen by a Thiocoumarin-Based Ratiometric Fluorescent Probe

The development of fluorescent probes derived from thiocarbonyl compounds for reactive oxygen species has been actively pursued in recent years. However, a better understanding of the optical response behaviors of thiocarbonyl compounds toward reactive oxygen species remains a challenge. Along with this, further studies to overcome the limitation of a single emission channel and aggregation-caused quenching features of thiocarbonyl-based fluorescent probes are highly desirable. Due to the important role of hypochlorite and singlet oxygen in biological processes and their common coexistence in living systems with frequent intertransformations, the design of a fluorescent probe that can recognize both hypochlorite and singlet oxygen is of great interest. Herein, a thiocarbonyl-based ratiometric fluorescent probe (Fcoum-S) for simultaneous detection of hypochlorite and singlet oxygen in aqueous solution and living cells was designed and synthesized. Upon the addition of hypochlorite in Fcoum-S solution (phosphate-buffered saline, 10 mM, pH 7.4, 10% acetonitrile), a ratiometric fluorescence response was observed via a specific hypochlorite-promoted desulfurization reaction with a good linear relationship between the ratio of fluorescence intensities at 526 and 602 nm (I_526nm/I_602nm) and the hypochlorite concentrations (a low detection limit of 0.15 μM). Furthermore, upon green light irradiation, Fcoum-S was efficiently desulfurized to its oxo analogue (Fcoum-O) by in situ generated singlet oxygen, leading to a significant change in fluorescence. Fcoum-S could work well in an aqueous medium owing to the high reactivity of the thiocarbonyl group and the aggregation-induced emission characteristics. More importantly, Fcoum-S could target mitochondria and was successfully utilized for fluorescence imaging of mitochondrial hypochlorite/singlet oxygen in live cells. This work provides a molecular design guideline for further exploring thioketone derivatives as fluorescent probes.