Figure 1.

Gating strategy used in multiparametric flow cytometry analysis to detect different subsets of monocytes. Doublets exclusion on FSC-A/FSC-H dot plot (A). Live cells on FSC-A/SSC-A (B). Monocytes identification as CD45⁺/SSC intermediate cells (C). Invert gate and exclusion of cells expressing CD24⁺, granulocytes expressing CD16^+high, and lymphocytes expressing CD56⁺ and CD2⁺ (D–G). Exclusion of residual cells CD14⁻ CD16⁻ (H). Characterization of human monocyte subsets into CD14⁺/CD16⁻ (MO1, conventional), CD14⁺/CD16⁺ (MO2, intermediate), and CD14^low/CD16⁺ (MO3, unconventional) subsets (I). Cell hierarchy (J).