Journal: STAR Protocols
Article Title: Protocol to phenotype and quantify mycobacteria-specific myeloid cells from human airways by mass cytometry
doi: 10.1016/j.xpro.2024.103463
Figure Lengend Snippet: Optimizing antibody staining and resolution in BAL cells (A) Antibody staining was tested on fresh PBMCs followed by MaxPar Fix PERM incubation with Ir 191/193 (no fix) after fixation with 2% and 4% paraformaldehyde (PFA) or BD FACS lyse buffer followed by MaxPar Fix PERM incubation with Ir 191/193 for antibodies CD4 clone RPA-T4 (row 1), CD56 clone NCAM16.2 (row 2), CD16 clone 3G8 and CD11b clone ICRF44 (row 3). Staining after fixative treatment reduced the resolution of these antibody clones compared to fresh cells. (B) Titrations of surface marker CD206 and intra-cellular marker IL1ß on BAL cells highlights the importance of titrating markers on both stimulated and unstimulated cells. (C) BAL or PBMCs were stained with the full antibody cocktail (stained) and compared to unstained controls (Ir 191/193 staining only) to determine true positive signal for markers CD19 and CD32 (row 1), CD19 and CD40 (row 2), CD11c and CD89 (row 3).
Article Snippet: Maxpar MCP9 antibody labeling kit, 112Cd , Standard BioTools , Cat# 201112A.
Techniques: Staining, Incubation, Clone Assay, Marker