应用举例 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45+ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatterhigh (SSChigh) cells, which correspond to low density PMN-MDSCs (region R5), CD14+cells (region R6) and SSClowCD14– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14+HLA-DR–. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing –CD33int.