Figure 6.

Leukocyte populations present in wounded back skin quantified by flow cytometry. Back skin wounds collected 5 days post wounding were digested with Liberase TL/Collagenase D cocktail to a single-cell suspension and the cells were labeled with antibodies to CD45, Lineage cocktail of antibodies (Lin) CD3, CD11b, CD11c, MHC-II, Ly6C, Ly6G, F4/80, NK1.1, CD127, ST2, and RORγt prior to analysis. Labeled cells were gated first on CD45 and then subsequently as shown. This multiplex gating strategy allows for efficient identification of seven innate-immune cell subtypes (Mϕ, macrophage; DCs, dendritic cells; NK, natural killer cells; ILCs, group 1, 2, and 3 innate lymphoid cells; neutrophil) from the same sample. This technique is highly suitable when needing to analyze immune populations in a localized, small sample such as a wound.