title: A Dual Functional Electroactive and Fluorescent Probe for Coupled Measurements of Vesicular Exocytosis with High Spatial and Temporal Resolution.
abstract: In this work, Fluorescent False Neurotransmitter 102 (FFN102), a synthesized analogue of biogenic neurotransmitters, was demonstrated to show both pH-dependent fluorescence and electroactivity. To study secretory behaviors at the single-vesicle level, FFN102 was employed as a new fluorescent/electroactive dual probe in a coupled technique (amperometry and total internal reflection fluorescence microscopy (TIRFM)). We used N13 cells, a stable clone of BON cells, to specifically accumulate FFN102 into their secretory vesicles, and then optical and electrochemical measurements of vesicular exocytosis were experimentally achieved by using indium tin oxide (ITO) transparent electrodes. Upon stimulation, FFN102 started to diffuse out from the acidic intravesicular microenvironment to the neutral extracellular space, leading to fluorescent emissions and to the electrochemical oxidation signals that were simultaneously collected from the ITO electrode surface. The correlation of fluorescence and amperometric signals resulting from the FFN102 probe allows real-time monitoring of single exocytotic events with both high spatial and temporal resolution. This work opens new possibilities in the investigation of exocytotic mechanisms.
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Liu
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Savy
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Maurin
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Grimaud
authors.affiliation: Laboratoire de Neurophotonique, CNRS UMR 8250, Université Paris Descartes, 45, rue des Saints-Pères, 75006, Paris, France.
authors.forename: François
authors.initials: F
authors.lastname: Darchen
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Damien
authors.initials: D
authors.lastname: Quinton
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Labbé
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Buriez
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Delacotte
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Lemaître
authors.affiliation: Ecole normale supérieure, PSL Research University, UPMC Univ Paris 06, CNRS, Département de Chimie, PASTEUR, 24, rue Lhomond, 75005, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, 75005, Paris, France.
authors.forename: Manon
authors.initials: M
authors.lastname: Guille-Collignon
mesh_terms: D002460:Cell Line; D055664:Electrochemical Techniques / Q000379:methods; D004566:Electrodes; D005089:Exocytosis; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008856:Microscopy, Fluorescence / Q000379:methods; D018377:Neurotransmitter Agents / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D018377:Neurotransmitter Agents
keywords: electrochemistry; exocytosis; indium tin oxides; neurotransmitters