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In Situ Fluorescence Tomography Enables a 3D Mapping of Enzymatic O 2 Reduction at the Electrochemical Interface

Abstract : Getting information about the fate of immobilized biomolecules and the evolution of their environment during turnover is a mandatory step towards bioelectrode optimization for effective use in biodevices. We demonstrate here the proof-ofprinciple characterization of the reactivity at an enzymatic electrode thanks to fluorescence confocal laser scanning microscopy (FCLSM) implemented in situ during the electrochemical experiment. The enzymatic O 2-reduction involves proton and electron transfers. Therefore, fluorescence variation of a pH-dependent fluorescent dye in the electrode vicinity enables the reaction visualization. Simultaneous collection of electrochemical and fluorescence signals gives valuable space-and time-resolved information. Once the technical challenges of such a coupling are overcome, in situ FCLSM affords a unique way to explore reactivity at the electrode surface and in the electrolyte volume. Unexpected features are observed, especially the pH evolution of the enzyme environment, which is also indicated by a characteristic concentration profile within the diffusion layer. This coupled approach gives also access to a cartography of the electrode surface response (i.e. heterogeneity), which cannot be obtained solely by an electrochemical mean.
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Submitted on : Thursday, December 3, 2020 - 1:57:24 PM
Last modification on : Wednesday, November 3, 2021 - 4:04:28 AM
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Bastien Tassy, Alice Dauphin, Hiu Mun Man, Hugo Le Guenno, Elisabeth Lojou, et al.. In Situ Fluorescence Tomography Enables a 3D Mapping of Enzymatic O 2 Reduction at the Electrochemical Interface. Analytical Chemistry, American Chemical Society, 2020, 92 (10), pp.7249-7256. ⟨10.1021/acs.analchem.0c00844⟩. ⟨hal-02936237⟩



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