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Updated: May 23, 2025
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Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
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Protocol for phenotyping mouse myeloid and lymphoid cells by mass cytometry
Laura Morin1, Brice Autier2, Patrice Hemon3
1University Rennes, INSERM, EHESP, IRSET (Institut de recherche en santé, environnement et travail) - UMR_S 1085, 35000 Rennes, France.
STAR Protocols
|March 11, 2025
Summary
This study details a protocol for immunophenotyping mouse immune cells using Cytometry by Time-Of-Flight (CyTOF). The method is applicable to various tissues and conditions, comparing fresh and cryopreserved samples.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Cytometry by Time-Of-Flight (CyTOF) is an advanced flow cytometry method.
- Metal-tagged antibodies enable high-dimensional immunophenotyping.
- Accurate immune cell profiling is crucial in health and disease.
Purpose of the Study:
- To present a detailed protocol for mouse immune cell phenotyping using CyTOF.
- To cover myeloid and lymphoid cell populations from multiple tissues.
- To compare sample processing of fresh versus cryopreserved tissues.
Main Methods:
- Protocol development for antibody labeling and titration.
- Standardized tissue dissociation techniques for lung, spleen, and lavage samples.
- Staining procedures for CyTOF analysis.
Main Results:
- Established a robust CyTOF protocol for diverse mouse immune cell subsets.
- Demonstrated comparability of staining from fresh and cryopreserved tissues.
- Provided a comprehensive guide for immunophenotyping.
Conclusions:
- The presented CyTOF protocol offers a reliable method for comprehensive mouse immune cell analysis.
- This protocol facilitates research in both healthy and pathologic conditions.
- The method is adaptable for various tissue types and sample preservation states.
