Fig. 3: Combined image analysis and digital biomarker profiling exemplified for diffuse and focal immune infiltration regions in one HGSC patient.

Immune infiltration of (a) diffuse and (b) focal patterns was identified in different cores of the same tumor (patient P387). Upper panels: Boundary of ROI (left) and segmentation into immune and tumor AOIs (right) in the DSP analysist (red=PanCk, green=CD45, blue=Syto13). Bottom panels: corresponding FIJI pre-processed ROI image (left), and DL-based segmentation and cell classification in QuPath (right). Right panels (larger image): Graph networks overlayed grey scale Syto13 ROI image. Nodes are colored in pink for tumor and green for immune. Connections within a 30-pixel (=12 µm) distance from centre of each node are displayed and were used to calculate spatial statistics. Scale bars represent 20 µm. c Normalized counts of selected biomarkers quantified in immune AOIs of diffuse and focal immune infiltration regions in tumor of P387. CD45 was similar in both AOIs. Diffuse immune infiltration was higher in CD3, CD8, CD44, and GZMB, and focal immune infiltration was higher in CD68, CD14 and CD20. d Spatial statistics derived from image analysis of the P387 tumor. Immune cell ratio was similar in diffuse and focal immune infiltration. Gdc was higher and tumor ccr lower in diffuse compared to focal infiltration. *the same y-axis scale are used for different spatial parameters; ccr scores, gdc scores and immune/(tumor+immune) ratio, respectively. ccr values have been scaled with a factor of 0.1 to enable visualization in the same plot.