During incubation, make the antibody cocktails for the “all stain” master mix (to stain samples and all-stained control) as well as any FMOs or SCCs you may be running. FMOs & SCCs can be prepared in 8-tube PCR strips. All antibody mixes should be prepared in a concentrated stain buffer: Fc Block (BioLegend) + Monocyte Block (BioLegend) + Brilliant Stain Buffer (BD Biosciences). Then just before staining cells, use flow wash buffer to bring all stain mixes to correct volume to stain all samples and controls. Flow wash buffer: 1X PBS w/o Ca2+ or Mg2+ (Gibco 14190-144), 1% w/v bovine serum albumin (Sigma A9647), 1 mM EDTA pH 8.0 (Invitrogen 15575-038). A suggested panel for staining stromal, immune, and vascular cells is included: