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

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.

Electrochemical monitoring of the fluorescence emission of tetrazine and bodipy dyes using total internal reflection fluorescence microscopy coupled to electrochemistry.

A very sensitive technique where an electrochemical cell is coupled to a total internal reflection fluorescence microscopy setup is described and applied for the first time to the electrochemical monitoring of the fluorescence of organic dyes in solution. It is shown that this setup basically allows both spatial and time resolution for the recorded fluorescence signal as a function of the electrode potential: indeed the variations of the emission intensity are recorded within the diffusion layer for a classical cyclic voltammetry or chronoamperometry experiment inducing the redox conversion of an emissive form into a non emissive one (and conversely). Simultaneously, the variations of the emissive state lifetime are measured to discriminate between a mechanism involving only the conversion into a non emissive form from one involving a quenching between the emitter and the electrogenerated species. The results concerning the investigation of the electrochemical monitoring of the fluorescence properties for two types of original dyes are presented, demonstrating the possibility to switch on and off the emission in a fully reversible way and to investigate in depth the mechanisms associated to this switch.