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

Ian H. Guldner, Samantha M. Golomb, Qingfei Wang, Emilia Wang, Siyuan Zhang

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

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).

Original languageEnglish (US)
Article number100537
JournalSTAR Protocols
Volume2
Issue number2
DOIs
StatePublished - Jun 18 2021
Externally publishedYes

Keywords

  • Cell isolation
  • Flow Cytometry/Mass Cytometry
  • Immunology
  • Neuroscience
  • RNA-seq
  • Sequencing
  • Single Cell

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Medicine(all)

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