Journal: STAR Protocols
Article Title: Protocol for phenotyping mouse myeloid and lymphoid cells by mass cytometry
doi: 10.1016/j.xpro.2025.103684
Figure Lengend Snippet: Proposed gating strategy for identifying key murine tissue cell populations in both peritoneal lavage and lung tissue Mouse lung cells were isolated and labeled with the panel of 37 markers proposed in this protocol. After cell acquisition using the Helios mass cytometer and data cleaning, cells were identified using a gating strategy on the OMIQ software. The CD45- cells and CD45+ immune cells were first identified. We then identify B cells (CD19 + ) and among them B1 and B2 subpopulations, T cells (CD3 + ) and among them LTCD8+ and LTCD4+ and notably LTreg (Foxp3 + ), Natural killers (NK1.1 + ), neutrophils (Ly6G + ) and eosinophils (CD24 + CD170 + ). The remaining population, called "other myeloid cells", consisted mainly of monocytes, macrophages and dendritic cells (DC). In peritoneal lavage, the two populations of conventional DC, cDC1 and cDC2, were identified among mDC (CD11c + ) based on CD172a and CD209a staining. After exclusion of plasmacytoid DC (pDC) (SiglecH + ), two subpopulations of peritoneal macrophages (PM) (CD68 + F4/80 + ) were identified, the small PM (SPM)(I-A/I-E hi and CCR hi ) and the large PM (LPM). Finally, we identified classical monocytes based on Ly6c hi and CD43 lo staining. In lung tissue, we first identified alveolar macrophages (AM) (CD64 + CD68 + ) and after DC exclusion, we identified interstitial macrophages (IM) subpopulations (CD11b + I-A/I-E + ) and classical monocytes (Ly6c hi and CD43 lo ). Among CD11c + dendritic cells, we next identified plasmacytoid DC (pDC) (SiglecH + ) and the two subpopulations of conventional DC named cDC1 (CD103 + ) and cDC2 (CD172a + ).
Article Snippet: Anti-mouse CD19 (6D5)-149Sm-100 tests , Standard BioTools , Cat#3149002B; RRID: AB_2814679.
Techniques: Isolation, Labeling, Cytometry, Software, Staining
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