Combined two-photon imaging, electrophysiological, and anatomical investigation of the human neocortex in vitro

Bálint Péter Kerekes; Kinga Toth; Attila Kaszás; Balázs Chiovini; Zoltán Szadai; Gergely Szalay; Denes Palfi; Attila Bagó; Klaudia Spitzer; Balázs Rózsa; István Ulbert; Lucia Wittner

doi:10.1117/1.NPh.1.1.011013

11 September 2014 Combined two-photon imaging, electrophysiological, and anatomical investigation of the human neocortex in vitro

Bálint Péter Kerekes, Kinga Toth, Attila Kaszás, Balázs Chiovini, Zoltán Szadai, Gergely Szalay, Denes Palfi, Attila Bagó, Klaudia Spitzer, Balázs Rózsa, István Ulbert, Lucia Wittner

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Neurophotonics, Vol. 1, Issue 1, 011013 (September 2014). https://doi.org/10.1117/1.NPh.1.1.011013

Abstract

Spontaneous synchronous population activity (SPA) can be detected by electrophysiological methods in cortical slices of epileptic patients, maintained in a physiological medium in vitro. In order to gain additional spatial information about the network mechanisms involved in the SPA generation, we combined electrophysiological studies with two-photon imaging. Neocortical slices prepared from postoperative tissue of epileptic and tumor patients were maintained in a dual perfusion chamber in a physiological incubation medium. SPA was recorded with a 24-channel extracellular linear microelectrode covering all neocortical layers. After identifying the electrophysiologically active regions of the slice, bolus loading of neuronal and glial markers was applied on the tissue. SPA-related Ca ²+ transients were detected in a large population of neighboring neurons with two-photon microscopy, simultaneous with extracellular SPA and intracellular whole-cell patch-clamp recordings. The intracellularly recorded cells were filled for subsequent anatomy. The cells were reconstructed in three dimensions and examined with light- and transmission electron microscopy. Combining high spatial resolution two-photon Ca ²+ imaging techniques and high temporal resolution extra- and intracellular electrophysiology with cellular anatomy may permit a deeper understanding of the structural and functional properties of the human neocortex.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Bálint Péter Kerekes, Kinga Toth, Attila Kaszás, Balázs Chiovini, Zoltán Szadai, Gergely Szalay, Denes Palfi, Attila Bagó, Klaudia Spitzer, Balázs Rózsa, István Ulbert, and Lucia Wittner "Combined two-photon imaging, electrophysiological, and anatomical investigation of the human neocortex in vitro," Neurophotonics 1(1), 011013 (11 September 2014). https://doi.org/10.1117/1.NPh.1.1.011013

Published: 11 September 2014

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