Sainsbury Wellcome Centre for Neural Circuits and Behaviour, University College London, London, United Kingdom.
PLoS One. 2022 Aug 11;17(8):e0271832. doi: 10.1371/journal.pone.0271832. eCollection 2022.
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.
本方案是为准备幼年成年小鼠中脑用于电生理实验的急性冠状切片而制定的实用指南。它描述了两套不同的溶液及其各自的孵育策略，以及两种替代的脑提取方法：在终末异氟醚麻醉下断头和在终末异氟醚麻醉下用人工脑脊液进行心脏内灌注。切片可以从野生型小鼠以及经过基因修饰或转染病毒构建体以标记细胞亚群的小鼠中制备。该制备可用于结合药理学、光遗传学和化学遗传学操作以及钙成像来研究中脑神经元的电生理特性；随后可以进行形态重建、免疫组织化学或单细胞转录组学分析。该方案还提供了详细的材料和试剂清单，包括低成本且易于组装的 3D 打印切片回收室的设计、解决导致切片质量不佳的常见问题的一般建议，以及一些确保良好维护贴附式细胞电极记录设备的建议。
