Enhancing the Study of Quantal Exocytotic Events: Combining Diamond Multi-Electrode Arrays with Amperometric PEak Analysis (APE) an Automated Analysis Code

Giulia Tomagra; Alice Re; Veronica Varzi; Pietro Aprà; Adam Britel; Claudio Franchino; Sofia Sturari; Nour-Hanne Amine; Remco H S Westerink; Valentina Carabelli; Federico Picollo

doi:10.3390/bios13121033

Enhancing the Study of Quantal Exocytotic Events: Combining Diamond Multi-Electrode Arrays with Amperometric PEak Analysis (APE) an Automated Analysis Code

Biosensors (Basel). 2023 Dec 16;13(12):1033. doi: 10.3390/bios13121033.

Affiliations

¹ Department of Drug and Science Technology, NIS Interdepartmental Centre, University of Torino, Corso Raffaello 30, 10125 Torino, Italy.
² Department of Physics, NIS Interdepartmental Centre, University of Torino and Italian Institute of Nuclear Physics, Via Giuria 1, 10125 Torino, Italy.
³ Neurotoxicology Research Group, Division of Toxicology, Institute for Risk Assessment Sciences (IRAS), Faculty of Veterinary Medicine, Utrecht University, P.O. Box 80.177, NL-3508 TD Utrecht, The Netherlands.

Abstract

MicroGraphited-Diamond-Multi Electrode Arrays (μG-D-MEAs) can be successfully used to reveal, in real time, quantal exocytotic events occurring from many individual neurosecretory cells and/or from many neurons within a network. As μG-D-MEAs arrays are patterned with up to 16 sensing microelectrodes, each of them recording large amounts of data revealing the exocytotic activity, the aim of this work was to support an adequate analysis code to speed up the signal detection. The cutting-edge technology of microGraphited-Diamond-Multi Electrode Arrays (μG-D-MEAs) has been implemented with an automated analysis code (APE, Amperometric Peak Analysis) developed using Matlab R2022a software to provide easy and accurate detection of amperometric spike parameters, including the analysis of the pre-spike foot that sometimes precedes the complete fusion pore dilatation. Data have been acquired from cultured PC12 cells, either collecting events during spontaneous exocytosis or after L-DOPA incubation. Validation of the APE code was performed by comparing the acquired spike parameters with those obtained using Quanta Analysis (Igor macro) by Mosharov et al.

Keywords: PC12 cell line; amperometry; diamond; dopamine; exocytosis; multi-electrode array.

MeSH terms

Animals
Chromaffin Cells* / physiology
Diamond
Exocytosis / physiology
Hominidae*
Microelectrodes
Rats

Substances

Diamond

Grants and funding

The “Trapezio” project was funded by the Compagnia di San Paolo (2022–2024, n. 2021.2236), the “QuantDia” project funded by the Italian Ministry for Education, University and Research (MIUR) within the “FISR 2019” program, and the “LasIonDef” project was funded by the European Research Council under the “Marie Skłodowska-Curie Innovative Training Networks” program (GA n. 956387).