Figure 1.

Quantitative multiplexed evaluation of macrophage polarization, location, and density in colorectal cancer tissue microarrays. (A) Multiplex immunofluorescence image of a representative tumor core. (B-D) Machine learning based image processing, including tissue category classification (B), cell segmentation (C), and cell type classification (D) to identify macrophages in tumor intraepithelial and stromal regions. (E) Examples of macrophages with M1-like, M2-like, and mixed expression phenotypes. (F) Assignment of polarization state for each macrophage across the M1:M2 spectrum using a scaled, four marker index. Plot shows the distribution of M1:M2-index values for all macrophages (N=367,264) across 931 tumors. Macrophages present within the two-sided 30% tails of the graph were classified as M1-like or M2-like for the main analysis. (G) Distribution of overall macrophage density and M1:M2 density ratio across 931 tumors. Scale bars indicate 100 μm (A) or 10 μm (E).