Pavón Arocas, Oriol;
Branco, Tiago;
(2022)
Preparation of acute midbrain slices containing the superior colliculus and periaqueductal Gray for patch-clamp recordings.
PLoS One
, 17
(8)
, Article e0271832. 10.1371/journal.pone.0271832.
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Abstract
This protocol is a practical guide for preparing acute coronal slices from the midbrain of young adult mice for electrophysiology experiments. It describes two different sets of solutions with their respective incubation strategies and two alternative procedures for brain extraction: decapitation under terminal isoflurane anaesthesia and intracardial perfusion with artificial cerebrospinal fluid under terminal isoflurane anaesthesia. Slices can be prepared from wild-type mice as well as from mice that have been genetically modified or transfected with viral constructs to label subsets of cells. The preparation can be used to investigate the electrophysiological properties of midbrain neurons in combination with pharmacology, opto- and chemogenetic manipulations, and calcium imaging; which can be followed by morphological reconstruction, immunohistochemistry, or single-cell transcriptomics. The protocol also provides a detailed list of materials and reagents including the design for a low-cost and easy to assemble 3D printed slice recovery chamber, general advice for troubleshooting common issues leading to suboptimal slice quality, and some suggestions to ensure good maintenance of a patch-clamp rig.
| Type: | Article |
|---|---|
| Title: | Preparation of acute midbrain slices containing the superior colliculus and periaqueductal Gray for patch-clamp recordings |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1371/journal.pone.0271832 |
| Publisher version: | https://doi.org/10.1371/journal.pone.0271832 |
| Language: | English |
| Additional information: | © 2022 Pavón Arocas, Branco. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
| UCL classification: | UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > The Sainsbury Wellcome Centre UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10153745 |
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