Fig. 1

Study design overview and quality control. A Representative multiplex immunostained images of samples subjected to the 3 designed panels for the same patient. An autofluorescence-simulated HE histopathology image of the M panel is also shown. Scale bar, 200 μm. B Image processing workflow implemented by inForm software. The input image a was subjected to spectral unmixing b, tissue segmentation into epithelial and TME compartments (red: epithelium; green: TME) c, cell segments d and phenotyping e. C Correlation dot plot and density plot of the inferred purity. The correlation coefficient and p value are shown in the text. D H-score of the assessed molecules comparing the epithelial and TME compartments from the same patient. Bars show the median with interquartile range. The paired Wilcoxon test was applied, with ns > 0.05 *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001