Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes

STAR Protoc. 2021 May 13;2(2):100537. doi: 10.1016/j.xpro.2021.100537. eCollection 2021 Jun 18.

Abstract

High dimensional compositional and transcriptional profiling of heterogeneous brain-infiltrating leukocytes can lead to novel biological and therapeutic discoveries. High-quality single-cell leukocyte preparations are a prerequisite for optimal single cell profiling. Here, we describe a protocol for epitope and RNA-preserving dissociation of adult mouse brains and subsequent leukocyte purification and staining, which is adaptable to homeostatic and pathogenic brains. Leukocyte preparation following this protocol permits exquisite single-cell surface protein and RNA profiling in applications including CyTOF and CITE-seq. For complete details on the use and execution of this protocol, please refer to Guldner et al. (2020) and Golomb et al. (2020).

Keywords: Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Neuroscience; RNA-seq; Sequencing; Single Cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / pathology*
  • Cell Separation / methods*
  • Cells, Cultured
  • Epitopes / genetics*
  • Female
  • Leukocytes* / cytology
  • Leukocytes* / metabolism
  • Male
  • Mice
  • Single-Cell Analysis / methods*
  • Transcriptome / genetics*

Substances

  • Epitopes