Coupling Amperometry and Total Internal Reflection Fluorescence Microscopy at ITO Surfaces for Monitoring Exocytosis of Single Vesicles

Meunier, Anne; Jouannot, Ouardane; Fulcrand, Rémy; Fanget, Isabelle; Bretou, Marine; Karatekin, Erdem; Arbault, Stéphane; Guille, Manon; Darchen, François; Lemaître, Frédéric; Amatore, Christian

doi:10.1002/ange.201101148

Cited by 25 publications

(7 citation statements)

References 20 publications

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“…The recorded cellular signals demonstrate that an all‐carbon miniaturized device based on graphitic electrodes embedded in a diamond matrix can efficiently detect the quantal release of catecholamines from secretory cells. In particular, it is worth noting that the performance (amperometric sensitivity, signal‐to‐noise ratio, time resolution) of the prototypical μG‐SCD device presented in this work compare well not only with standard CFEs (see Table 1), but also with other well developed technologies at the state of the art, such as devices based on indium tin oxide (ITO) conductive glass,18–20 noble metals (Au, Pt, …︁)21, 22 and boron‐doped nanocrystalline diamond (B:NCD) 16. In conclusion, these results open promising perspectives for the realization of all‐carbon multielectrode miniaturized devices in artificial diamond (a material which is becoming available with increasing crystal quality at ever‐decreasing costs)23, 24 in which full advantage of the robustness, chemical stability, biocompatibility and transparency can be exploited to obtain multiparametric signals detection from cell networks.…”

Section: Mean Values and Standard Deviations Of The Main Parameters Dmentioning

confidence: 53%

A New Diamond Biosensor with Integrated Graphitic Microchannels for Detecting Quantal Exocytic Events from Chromaffin Cells

Picollo¹,

Gosso

Vittone³

et al. 2013

Advanced Materials

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The quantal release of catecholamines from neuroendocrine cells is a key mechanism which has been investigated with a broad range of materials and devices, among which carbon-based materials such as carbon fibers, diamond-like carbon, carbon nanotubes and nanocrystalline diamond. In the present work we demonstrate that a MeV-ion-microbeam lithographic technique can be successfully employed for the fabrication of an all-carbon miniaturized cellular bio-sensor based on graphitic micro-channels embedded in a single-crystal diamond matrix. The device was functionally characterized for the in vitro recording of quantal exocytic events from single chromaffin cells, with high sensitivity and signal-to-noise ratio, opening promising perspectives for the realization of monolithic all-carbon cellular biosensors.Submitted to 2

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Section: Mean Values and Standard Deviations Of The Main Parameters Dmentioning

confidence: 53%

A New Diamond Biosensor with Integrated Graphitic Microchannels for Detecting Quantal Exocytic Events from Chromaffin Cells

Picollo¹,

Gosso

Vittone³

et al. 2013

Advanced Materials

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“…For example, cell-attached recordings perturb membrane tension and the cytoskeleton strongly [134,135] and TIRFM detects release only from the substrateattached side of a cell. Although these perturbations may affect exocytosis to some degree, secretion [124] and pore properties [136][137][138] do not vary dramatically between the substrate-attached and bath-exposed sides of a cell (but see [139]).…”

Section: Discussion: a Unified Picture Of The Fusion Porementioning

confidence: 99%

Toward a unified picture of the exocytotic fusion pore

Karatekin

2018

FEBS Letters

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Neurotransmitter and hormone release involve calcium-triggered fusion of a cargo-loaded vesicle with the plasma membrane. The initial connection between the fusing membranes, called the fusion pore, can evolve in various ways, including rapid dilation to allow full cargo release, slow expansion, repeated opening-closing, and resealing. Pore dynamics determine the kinetics of cargo release and the mode of vesicle recycling, but how these processes are controlled are poorly understood. Previous reconstitutions could not monitor single pores, limiting mechanistic insight they could provide. Recently developed nanodisc-based fusion assays allow reconstitution and monitoring of single pores with unprecedented detail and hold great promise for future discoveries. They recapitulate various aspects of exocytotic fusion pores, but comparison is difficult because different approaches suggested very different exocytotic fusion pore properties, even for the same cell type. In this Review, I discuss how most of the data can be reconciled, by recognizing how different methods probe different aspects of the same fusion process. The resulting picture is that fusion pores have broadly distributed properties arising from stochastic processes which can be modulated by physical constraints imposed by proteins, lipids and membranes.

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“…This allowed the simultaneous monitoring of the vesicular release by amperometry and fluorescence. As a proof of concept, the dual detection was demonstrated with a stable clone of enterochromaffin BON cells (BC21) that expresses GFP‐tagged neuropeptide‐Y (NPY‐GFP) and releases the electroactive serotonin . However, the use of two independent probes (fluorescent and electroactive) prevented a quantitative analysis of the release due to a lack of control on the relative proportion of both probes within vesicles (simultaneous optical and electrochemical signals reached only ∼20 %).…”

Section: Introductionmentioning

confidence: 99%

A Fluorescent False Neurotransmitter as a Dual Electrofluorescent Probe for Secretory Cell Models

et al. 2019

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A dual electrofluorescent probe (FFN42) belonging to the fluorescent false neurotransmitter family was rationally designed for investigating cell secretion. This probe, which comprises a coumarin core with one amino and two hydroxy groups, is very promising due to its electroactive and fluorescent properties. The optimal excitation and emission wavelengths (380 nm and 470 nm respectively) make this probe adapted for use in fluorescence microscopy. FFN42 has a quantum yield of 0.18, a molar absorption coefficient of 12000 M−1cm−1 and pKa values of 5.4 and 6.7 for the hydroxy groups. The electroactivity of FFN42 was evidenced on carbon fiber and ITO electrodes at relatively low oxidation potentials (0.24 V and 0.45 V vs Ag/AgCl respectively). Epifluorescence observations showed that FFN42 accumulated into secretory vesicles of PC12 and N13 cells. Toxicity tests further revealed that FFN42 had no lethal effect on these cells. Amperometric data obtained on carbon fiber electrodes proved that the probe is released by N13 cells.

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Coupling Amperometry and Total Internal Reflection Fluorescence Microscopy at ITO Surfaces for Monitoring Exocytosis of Single Vesicles

Cited by 25 publications

References 20 publications

A New Diamond Biosensor with Integrated Graphitic Microchannels for Detecting Quantal Exocytic Events from Chromaffin Cells

A New Diamond Biosensor with Integrated Graphitic Microchannels for Detecting Quantal Exocytic Events from Chromaffin Cells

Toward a unified picture of the exocytotic fusion pore

A Fluorescent False Neurotransmitter as a Dual Electrofluorescent Probe for Secretory Cell Models

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