Premium
A Dual Functional Electroactive and Fluorescent Probe for Coupled Measurements of Vesicular Exocytosis with High Spatial and Temporal Resolution
Author(s) -
Liu Xiaoqing,
Savy Alexandra,
Maurin Sylvie,
Grimaud Laurence,
Darchen François,
Quinton Damien,
Labbé Eric,
Buriez Olivier,
Delacotte Jérôme,
Lemaître Frédéric,
GuilleCollig Ma
Publication year - 2017
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201611145
Subject(s) - amperometry , total internal reflection fluorescence microscope , exocytosis , fluorescence , chemistry , fluorescence microscope , indium tin oxide , biophysics , temporal resolution , vesicle , electrode , nanotechnology , electrochemistry , analytical chemistry (journal) , membrane , biochemistry , materials science , optics , chromatography , biology , physics
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.