Boronate-based fluorescent probes for imaging cellular hydrogen peroxide.
The syntheses, properties, and biological applications of the Peroxysensor family, a new class of fluorescent probes for hydrogen peroxide, are presented. These reagents utilize a boronate deprotection mechanism to provide high selectivity and optical dynamic range for detecting H2O2 in aqueous solution over similar reactive oxygen species (ROS) including superoxide, nitric oxide, tert-butyl hydroperoxide, hypochlorite, singlet oxygen, ozone, and hydroxyl radical. Peroxyresorufin-1 (PR1), Peroxyfluor-1 (PF1), and Peroxyxanthone-1 (PX1) are first-generation probes that respond to H2O2 by an increase in red, green, and blue fluorescence, respectively. The boronate dyes are cell-permeable and can detect micromolar changes in H2O2 concentrations in living cells, including hippocampal neurons, using confocal microscopy and two-photon microscopy. The unique combination of ROS selectivity, membrane permeability, and a range of available excitation/emission colors establishes the potential value of PR1, PF1, PX1, and related probes for interrogating the physiology and pathology of cellular H2O2.
Miller
Evan W
EW
Department of Chemistry, University of California, Berkeley, California 94720, USA.
Albers
Aaron E
AE
Pralle
Arnd
A
Isacoff
Ehud Y
EY
Chang
Christopher J
CJ
D001896:Boron Compounds / Q000737:chemistry*; D002460:Cell Line; D002477:Cells / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D006861:Hydrogen Peroxide / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D017785:Photons; D012996:Solutions / Q000737:chemistry; D014867:Water / Q000737:chemistry
D001896:Boron Compounds; D005456:Fluorescent Dyes; D012996:Solutions; D014867:Water; D006861:Hydrogen Peroxide