在胚胎到老年小鼠的急性脑片中进行全细胞膜片钳和染料加载，并结合批量 RNA 测序。
Combining whole-cell patch clamp and dye loading in acute brain slices with bulk RNA sequencing in embryonic to aged mice.
机构信息
Wellcome - Medical Research Council Cambridge Stem Cell Institute and Department of Veterinary Medicine, University of Cambridge, Cambridge CB2 0AW, United Kingdom.
Department of Physiology, Biomedical Centre, Faculty of Medicine, University of Iceland, Reykjavik, Iceland.
出版信息
STAR Protoc. 2021 Apr 9;2(2):100439. doi: 10.1016/j.xpro.2021.100439. eCollection 2021 Jun 18.
Single-cell electrophysiological recordings combined with dye loading and immunohistochemistry provide unparalleled single-cell resolution of cell physiology, morphology, location, and protein expression. When correlated with bulk RNA sequencing, these data can define cell identity and function. Here, we describe a protocol to prepare acute brain slices from embryonic and postnatal mice for whole-cell patch clamp, dye loading and post-hoc immunohistochemistry, and cell isolation for bulk RNA sequencing. While we focus on oligodendrocyte precursor cells, this protocol is applicable to other brain cells. For complete details on the use and execution of this protocol, please refer to Spitzer et al. (2019).
单细胞电生理记录与染料加载和免疫组织化学相结合，可提供无与伦比的单细胞分辨率，用于研究细胞生理学、形态学、位置和蛋白质表达。当与批量 RNA 测序相关联时，这些数据可以定义细胞的身份和功能。本文描述了一种从胚胎期和新生期小鼠大脑中制备急性脑切片的方案，用于全细胞膜片钳、染料加载和事后免疫组织化学以及批量 RNA 测序的细胞分离。虽然我们专注于少突胶质前体细胞，但该方案适用于其他脑细胞。有关此方案的使用和执行的完整详细信息，请参阅 Spitzer 等人（2019 年）。
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