Related Experiment Video
Updated: May 23, 2025
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
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.
Insights
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.
Abstract:
Cytometry by time of flight (CyTOF) is a flow cytometry-based technique using metal-tagged antibodies, allowing immunophenotyping. Here, we present a protocol for phenotyping mouse myeloid and lymphoid cells isolated from lung, spleen, and intraperitoneal lavages in healthy or pathologic conditions. We describe steps for antibody labeling and titration, tissue dissociations, and staining. We then detail procedures to compare staining obtained from fresh or cryopreserved tissues. For complete details on the use and execution of this protocol, please refer to Morin et al.1.
