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Combined scanning electrochemical and fluorescence microscopies using a tetrazine as a single redox and luminescent (electrofluorochromic) probe.

ABSTRACT: The possibility of using a single electroactive and luminescent molecule both as a redox mediator and as a fluorophore in an experiment combining in situ Scanning Electrochemical Microscopy (SECM) and epifluorescence microscopy was validated. The usual working modes of SECM, namely positive and negative feedback as well as generation-collection, were used and the fluorescence images, intensity and spectra were recorded for each configuration. The tip potential, tip-substrate distance and, in the case of a conducting substrate, the substrate potential are the parameters that are likely to control the fluorescence. It is shown that the tip can be used to switch on and off the luminescence and that the modulation amplitude maximum is sensitive to the nature of the substrate. Approach curves based on this fluorescence modulation amplitude can be obtained showing a higher sensitivity than the classical electrochemical ones.

SUBMITTER: Guerret-Legras L

PROVIDER: S-EPMC6050531 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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Combined scanning electrochemical and fluorescence microscopies using a tetrazine as a single redox and luminescent (electrofluorochromic) probe.

Guerret-Legras L L Audibert J F JF Dubacheva G V GV Miomandre F F

Chemical science 20180530 27

The possibility of using a single electroactive and luminescent molecule both as a redox mediator and as a fluorophore in an experiment combining <i>in situ</i> Scanning Electrochemical Microscopy (SECM) and epifluorescence microscopy was validated. The usual working modes of SECM, namely positive and negative feedback as well as generation-collection, were used and the fluorescence images, intensity and spectra were recorded for each configuration. The tip potential, tip-substrate distance and, ...[more]

PMID: 30079203

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Project description:The exact knowledge of hydrogel microstructure, mainly its pore topology, is a key issue in hydrogel engineering. For visualization of the swollen hydrogels, the cryogenic or high vacuum scanning electron microscopies (cryo-SEM or HVSEM) are frequently used while the possibility of artifact-biased images is frequently underestimated. The major cause of artifacts is the formation of ice crystals upon freezing of the hydrated gel. Some porous hydrogels can be visualized with SEM without the danger of artifacts because the growing crystals are accommodated within already existing primary pores of the gel. In some non-porous hydrogels the secondary pores will also not be formed due to rigid network structure of gels that counteracts the crystal nucleation and growth. We have tested the limits of true reproduction of the hydrogel morphology imposed by the swelling degree and mechanical strength of gels by investigating a series of methacrylate hydrogels made by crosslinking polymerization of glycerol monomethacrylate and 2-hydroxyethyl methacrylate including their interpenetrating networks. The hydrogel morphology was studied using cryo-SEM, HVSEM, environmental scanning electron microscopy (ESEM), laser scanning confocal microscopy (LSCM) and classical wide-field light microscopy (LM). The cryo-SEM and HVSEM yielded artifact-free micrographs for limited range of non-porous hydrogels and for macroporous gels. A true non-porous structure was observed free of artifacts only for hydrogels exhibiting relatively low swelling and high elastic modulus above 0.5 MPa, whereas for highly swollen and/or mechanically weak hydrogels the cryo-SEM/HVSEM experiments resulted in secondary porosity. In this contribution we present several cases of severe artifact formation in PHEMA and PGMA hydrogels during their visualization by cryo-SEM and HVSEM. We also put forward empirical correlation between hydrogel morphological and mechanical parameters and the occurrence and intensity of artifacts.

Dataset Information

Combined scanning electrochemical and fluorescence microscopies using a tetrazine as a single redox and luminescent (electrofluorochromic) probe.

Publications

Combined scanning electrochemical and fluorescence microscopies using a tetrazine as a single redox and luminescent (electrofluorochromic) probe.

Similar Datasets

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