Generation of single-cell and single-nuclei suspensions from embryonic and adult mouse brains

• Published in: STAR protocols 2023-03, Vol.4 (1), p.101944-101944, Article 101944
• Main Authors: Lee, Dongjin R., Zhang, Yajun, Rhodes, Christopher T., Petros, Timothy J.
• Format: Article
• Language: English
• Subjects: Cell Differentiation; Developmental biology; Flow Cytometry/Mass Cytometry; Neuroscience; Protocol; RNAseq; Sequencing; Single Cell
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101944

Efficient protocols to generate single-cell and single-nuclei suspensions are critical for the burgeoning field of single-cell/single-nuclei sequencing. Here we describe procedures to generate single-cell and single-nuclei suspensions from embryonic and adult mouse brains. This protocol can be modified for any brain region and/or neural cell type. For complete details on the use and execution of this protocol, please refer to Lee et al. (2022),1 Rhodes et al. (2022),2 Mahadevan et al. (2021),3 Ekins et al. (2020),4 and Wester et al. (2019).5 [Display omitted] •Obtain single-cell or single-nuclei suspensions from embryonic or adult mouse brains•Utilizes sorting to purify samples and select fluorescent cells/nuclei if needed•Describes different cell preparation strategies depending on downstream applications•Protocol is applicable to different ages and different brain regions Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Efficient protocols to generate single-cell and single-nuclei suspensions are critical for the burgeoning field of single-cell/single-nuclei sequencing. Here we describe procedures to generate single-cell and single-nuclei suspensions from embryonic and adult mouse brains. This protocol can be modified for any brain region and/or neural cell type.