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An endoplasmic reticulum (ER) targetable two-photon fluorescent probe termed ER-N3 is presented for the determination of hydrogen sulfide (H2S) in living cells and mice. The results demonstrate that ER-N3 was highly selective and sensitive to H2S. Confocal fluorescence imaging in living cells indicated that ER-N3 was accumulated in the endoplasmic reticulum and visualized exogenous and endogenous H2S production. Moreover, a burst of H2S upregulated by the nitric oxide donor sodium nitroprusside was monitored. Meanwhile, the increase of H2S ended following the addition of DL-propargylglycine, an inhibitor for cystathionine β-synthase and cystathionine γ-lyase that are the main sources of H2S. Importantly, the exogenous H2S was visualized in abdominal tissue of mice by utilizing two-photon fluorescence microscopy. The ER-N3 fluorescent probe may be an excellent tool to image H2S fluctuations in the endoplasmic reticulum and characterize the relationship between H2S and disease in vivo. [ABSTRACT FROM AUTHOR]

Hydrogen sulfide (H2S) is a critical gasotransmitter playing an important role in various physiological and pathological processes. The development of fluorescent probes for rapid, sensitive, and selective lysosomal H2S sensing has attracted much attention. Herein we report a rhodamine based probe MF‐N3 that can selectively accumulate in lysosomes and give turn‐on fluorescence in the presence of both H2S and protons. Results show that MF‐N3 has an exclusive response to H2S in the acidic lysosomal environment but does not respond in other organelles or in neutral environments. This strategy can circumvent the fluorescence interference from H2S of other sources, thus providing a high sensitivity and selectivity on lysosomal H2S imaging by fluorescence microscopy. This pH‐dependent lysosome‐targeting fluorescent probe has been successfully applied to lysosomal H2S imaging in living cells and is of great potential for applications to the environmental and biological systems. [ABSTRACT FROM AUTHOR]