Figure 2C shows a confocal image of a neutrophil-rich area of a mouse lung infected with Candida albicans and stained for NE (green, Millipore 48101) and H3 (red, ABIN 1735464) as well as for DNA (Hoechst 33342). Antigen retrieval was performed with R-Universal Buffer at 50°C. Using automatic Otsu thresholding, positive areas for both channels were depicted white, while areas below threshold were depicted black (Figures 2E,F). The overlap of both indicating the NET-positive area is shown in Figure 2G and superimposed on the tissue staining (Figure 2D).