Hydrogen sulfide (H2S) is a critical gaseous signaling molecule involved in the regulation of many bodily functions in the cardiovascular, immune, and nervous systems. The physiological outcomes of H2S in signaling pathways are complex, and advanced chemical and technological tools are required to deconvolute observed bioeffects and to strengthen our understanding of interconnectivities among these various pathways. Toward this goal, we have developed an azide-functionalized O-methylrhodol fluorophore, MeRho-Az, which exhibits a rapid &gt;1,000-fold fluorescence response when treated with H2S, is selective for H2S over other biological analytes, and has a detection limit of 86 nM. Importantly, the MeRho-Az scaffold is less susceptible to photoactivation than other commonly-used azide-based systems, which increases its potential application in imaging experiments where photobleaching may be an obstacle to effective analyte visualization. To demonstrate the efficacy of this probe for H2S detection in vivo, we used MeRho-Az in combination with light sheet fluorescence microscopy (LSFM) to visualize H2S in the intestinal tract of live zebrafish. This application represents the first demonstration of analyte-responsive 3D imaging with LSFM, highlighting the utility of combining fluorescence detection strategies with live imaging technologies for investigating chemical signaling in complex multicellular systems.

Bright fluorescent probe enables analyte responsive, 3D imaging of H2S in live zebrafish using light-sheet fluorescence microscopy

251st National Meeting (2016)

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