A ratiometric two-photon fluorescent probe for imaging hydrogen sulfide in lysosomes

Hydrogen sulfide (H2S) is a kind of gaseous signalling molecule that plays pivotal role in various biological processes. So far, it is still a challenge to develop convenient and reliable methods for H2S detection in lysosomes. Herein, we developed a novel ratiometric two-photon fluorescent probe LR-H2S for imaging H2S in lysosomes. Upon the addition of H2S (using Na2S as a donor) to LR-H2S in buffer solution, the azide group is reduced to amino group and subsequently the carbamate ester is cleaved by 1,6-elimination, resulting in a fluorescence emission increase at 541 nm and a concomitant emission decrease at 475 nm. Under two-photon excitation of 840 nm, an 80-fold fluorescence ratio (F-541/F-475) enhancement was observed with a wide linear range of 25-2500 mu M. The detection limit was calculated to be 0.70 mu M based on 3 sigma/k method, indicating that the probe can detect H2S with a high sensitivity. The probe also shows excellent selectivity toward H2S among other biological interference species and features with low cytotoxicity and favorable two-photon properties. Furthermore, LR-H2S can easily localize in lysosomes and vividly illuminate endogenous/exogenous H2S level and distribution in lysosomes of living SGC-7901 cells.