title: Fluorescence-enabled electrochemical microscopy with dihydroresorufin as a fluorogenic indicator.
abstract: Recently, we introduced a new electrochemical imaging technique called fluorescence-enabled electrochemical microscopy (FFEM). The central idea of FEEM is that a closed bipolar electrode is utilized to electrically couple a redox reaction of interest to a complementary fluorogenic reaction converting an electrochemical signal into a fluorescent signal. This simple strategy enables one to use fluorescence microscopy to observe conventional electrochemical processes on very large electrochemical arrays. The initial demonstration of FEEM focused on the use of a specific fluorogenic indicator, resazurin, which is reduced to generate highly fluorescent resorufin. The use of resazurin has enabled the study of analyte oxidation reactions, such as the oxidation of dopamine and H2O2. In this report, we extend the capability of FEEM to the study of cathodic reactions using a new fluorogenic indicator, dihydroresorufin. Dihydroresorufin is a nonfluorescent molecule, which can be electrochemically oxidized to generate resorufin. The use of dihydroresorufin has enabled us to study a series of reducible analyte species including Fe(CN)6(3-) and Ru(NH3)6(3+). Here we demonstrate the correlation between the simultaneously recorded fluorescence intensity of resorufin and electrochemical oxidation current during potential sweep experiments. FEEM is used to quantitatively detect the reduction of ferricyanide down to a concentration of approximately 100 μM on a 25 μm ultramicroelectrode. We also demonstrate that dihydroresorufin, as a fluorogenic indicator, gives an improved temporal response and significantly decreases diffusional broadening of the signal in FEEM as compared to resazurin.
authors.affiliation: Department of Chemistry, University of Washington , Seattle, Washington 98195-1700, United States.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Oja
authors.forename: Joshua P
authors.initials: JP
authors.lastname: Guerrette
authors.forename: Michelle R
authors.initials: MR
authors.lastname: David
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
mesh_terms: D055664:Electrochemical Techniques / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010078:Oxazines / Q000737:chemistry*; D010084:Oxidation-Reduction
chemical_list: D005456:Fluorescent Dyes; D010078:Oxazines; C568681:dihydroresorufin