Tissular H2S plays important role in neuroprotection, cardioprotection and angiogenesis. Herein, we develop a new fluorescent probe (Z)-4-(2-(6-(2,4-dinitrophenoxy)naphthalen-2-yl)vinyl)-1-methylpyridinium (EMII-H2S) for tissular imaging of H2S. EMII-H2S allows for sensitive response toward H2S under the excitation of both one-photon (410 nm) and two-photon (750 nm). Asymmetric electron structure contributes to the large stokes shift of 160 nm. With the better absorbance (39000 M−1cm−1), quantum yield (0.201) as well as the two-photon absorption cross-section (119 GM), EMII-H2S provides the higher brightness in response to H2S under either one-photon (εΦ = 7638) or two-photon modes (δΦ = 23.8), which compare favorably with many other highly bright fluorescent probes. EMII-H2S is applied in imaging of cellular endogenous H2S and tissue imaging at depths of 160 μm–200 μm in mouse liver. With the satisfactory properties, EMII-H2S is used to estimate the dynamic H2S release during metabolism of two type of H2S donors, demonstrating that the thiovaline provided the long-lasting H2S production contrary to the rapid burst caused by NaHS. EMII-H2S proves to be an effective tool for imaging tissular H2S, which is of great significance for the visual analysis of gasotransmitters in tissue.