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High-speed imaging of glutamate release with genetically encoded sensors

Dürst, Céline D., Wiegert, J. Simon, Helassa, Nordine, Kerruth, Silke, Coates, Catherine, Schulze, Christian, Geeves, Michael A., Török, Katalin, Oertner, Thomas G. (2019) High-speed imaging of glutamate release with genetically encoded sensors. Nature Protocols, 14 . pp. 1401-1424. ISSN 1754-2189. (doi:10.1038/s41596-019-0143-9)

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Abstract

The strength of an excitatory synapse depends on its ability to release glutamate and on the density of

synaptic transmission at the level of cleft glutamate to investigate properties of the release machinery in

investigating synaptic release during 100 Hz trains. Here we describe the detailed procedures for design

hippocampal cultures, and imaging of evoked glutamate transients with two-photon laser scanning

localize the vesicle fusion site with a precision exceeding the optical resolution of the microscope. By

iGluSnFR transients. The typical time frame for these experiments is 30 min per synapse.

Item Type: Article
DOI/Identification number: 10.1038/s41596-019-0143-9
Uncontrolled keywords: genetically-encoded glutamate indicator, GEGI, glutamate, two-photon imaging, two-photon microscopy, synaptic transmission, stopped-flow, iGluSnFR, hippocampal culture, rat, pyramidal cell, CA1, excitatory synapse, multivesicular release, organotypic culture, single-cell electroporation
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Michael Geeves
Date Deposited: 25 Mar 2019 15:53 UTC
Last Modified: 14 Oct 2019 23:00 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/73197 (The current URI for this page, for reference purposes)
Geeves, Michael A.: https://orcid.org/0000-0002-9364-8898
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