Work

Protocol for phenotyping mouse myeloid and lymphoid cells by mass cytometry

Year: 2025

Type: article

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 • Protocol to phenotype mouse myeloid and lymphoid cells by mass cytometry • Antibody labeling to heavy metals and titration of a 37-antibody panel • Steps to prepare single-cell suspension from lung and spleen tissues • Cell staining, sample acquisition, data cleaning, and cell gating strategy Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.103684