Protocol for murine multi-tissue deep immunophenotyping using a 40-color full-spectrum flow cytometry panel

STAR Protoc. 2024 Dec 20;5(4):103492. doi: 10.1016/j.xpro.2024.103492. Epub 2024 Dec 12.

Abstract

The innate and adaptive immune systems, though often studied separately, interact deeply and respond to stimuli simultaneously, with leukocytes displaying a range of pro- to anti-inflammatory phenotypes. This protocol details a procedure for characterizing murine innate and adaptive immune phenotypes using a 40-color full-spectral flow cytometry panel. We describe steps for organ collection, sample preparation, immunofluorescent staining, and acquisition to reproducibly and cost-effectively study tissue-resident leukocytes, their subpopulations, and inflammatory status in various organs.

Keywords: antibody; cell isolation; flow cytometry; immunology.

MeSH terms

  • Adaptive Immunity / immunology
  • Animals
  • Flow Cytometry* / methods
  • Immunity, Innate / immunology
  • Immunophenotyping* / methods
  • Leukocytes / cytology
  • Leukocytes / immunology
  • Mice