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Combining whole-cell patch clamp and dye loading in acute brain slices with bulk RNA sequencing in embryonic to aged mice

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. Her...

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Detalles Bibliográficos
Autores principales: Kamen, Yasmine, Káradóttir, Ragnhildur Thóra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8055713/
https://www.ncbi.nlm.nih.gov/pubmed/33899020
http://dx.doi.org/10.1016/j.xpro.2021.100439
Descripción
Sumario: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).