Azide-based fluorescent probes: imaging hydrogen sulfide in living systems.
Hydrogen sulfide is a redox active sulfur species that is endogenously generated in mammalian systems as an antioxidant and signaling molecule to support cellular function. The fundamental and ubiquitous actions of hydrogen sulfide demand sensitive and specific methods to track this biomolecule as it is produced within living organisms with temporal and spatial regulation. In this context, the hydrogen sulfide-mediated reduction of an azide to an amine is a useful method for organic synthesis, and this reaction has successfully been exploited to yield biocompatible fluorescent probes for hydrogen sulfide detection in vitro and in cells. This chapter provides protocols and guidelines for applying azide-based fluorescence probes to detecting hydrogen sulfide in living systems, including a protocol that was used to detect endogenous hydrogen sulfide in living single cells using a confocal microscope.
Department of Chemistry, University of California, Berkeley, California, USA.
Vivian S
VS
Lin
Department of Chemistry, Southern Methodist University, Dallas, Texas, USA; Center for Drug Discovery, Design, and Delivery (CD4), Southern Methodist University, Dallas, Texas, USA. Electronic address: alippert@smu.edu.
Alexander R
AR
Lippert
Department of Chemistry, University of California, Berkeley, California, USA; Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley, California, USA; Howard Hughes Medical Institute, Chevy Chase, Maryland, USA. Electronic address: chrischang@berkeley.edu.
Christopher J
CJ
Chang
D001386:Azides / Q000737:chemistry*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005456:Fluorescent Dyes / Q000737:chemistry*; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D006862:Hydrogen Sulfide / Q000737:chemistry* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D013194:Staining and Labeling; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D001386:Azides; D005456:Fluorescent Dyes; D042461:Vascular Endothelial Growth Factor A; D006862:Hydrogen Sulfide
Azide reduction; Fluorescence; Hydrogen sulfide; Microscopy; Reaction-based sensing; Redox biology