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Related Experiment Video

Updated: Jun 13, 2025

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

Pierre Lemaitre1, Chantal Mathieu1, Conny Gysemans1

  • 1Clinical and Experimental Endocrinology (CEE), CHROMETA, KU Leuven, Campus Gasthuisberg O&N1bis, Leuven, Belgium.

STAR Protocols
|December 13, 2024
PubMed
Summary

This study presents a 40-color flow cytometry method to analyze both innate and adaptive immune cells in mice. The protocol enables comprehensive characterization of leukocyte phenotypes and inflammatory status across different organs.

Keywords:
antibodycell isolationflow cytometryimmunology

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Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Innate and adaptive immunity are intricately linked, responding together to stimuli.
  • Leukocytes exhibit diverse phenotypes, ranging from pro-inflammatory to anti-inflammatory.
  • Understanding these interactions is crucial for immune system research.

Purpose of the Study:

  • To detail a reproducible and cost-effective protocol for characterizing murine immune phenotypes.
  • To enable simultaneous analysis of innate and adaptive immune system components.
  • To facilitate the study of tissue-resident leukocytes and their inflammatory status.

Main Methods:

  • Development of a 40-color full-spectral flow cytometry panel.
  • Standardized procedures for organ collection and sample preparation.
  • Immunofluorescent staining and data acquisition techniques.

Main Results:

  • A comprehensive protocol for characterizing diverse leukocyte subpopulations.
  • Methodology for assessing pro- to anti-inflammatory phenotypes.
  • Application to study immune status in various murine organs.

Conclusions:

  • The described flow cytometry panel provides a robust method for immune phenotyping.
  • This protocol allows for cost-effective and reproducible analysis of complex immune responses.
  • Facilitates deeper understanding of tissue-specific immune cell behavior and inflammation.