A FRET-based ratiometric fluorescent probe for hydrogen polysulfide detection in living cells and zebrafish

Hydrogen polysulfide (H₂S_n, n > 1) is an important active sulfur molecule (RSS) in organisms, which have been considered to be involved in redox signaling and cytoprotective processes. In this work, in order to quickly and accurately detect H₂S_n in biosystems, 2-fluoro-5-nitrobenzoic ester was used as the response moiety for H₂S_n, and the FRET strategy was adopted to effectively connect the donor (6-hydroxy-2-naphthoic acid) and acceptor (4-substituted-1,8-naphthalimide) to construct a new ratiometric H₂S_n fluorescent probe NPNA-H₂S_n. NPNA-H₂S_n exhibited a more than ∼ 8.0-fold ratio enhancement towards H₂S_n at I₄₅₀/I₅₂₆ and a very high sensitivity with a very low detection limit of 40.3 nM. Impressive, NPNA-H₂S_n was further used for fluorescence imaging of H₂S_n in living cells and zebrafish, which showed high-clear ratiometric images. Therefore, we have demonstrated that NPNA-H₂S_n could be applied for ratiometric images of endogenous H₂S_n in living biosystems and provide a powerful molecular tool for evaluating the physiological and pathological functions of H₂S_n.