CD3 and CD8 positivity in single cells was determined by Gaussian mixture modeling45, with the median positive fraction for each marker (CD3 or CD8) in each region (CT or IM) across all 40 CRC cases used as the cutoff for assigning a subscore of 0 or 1; the sum of the four subscores was used as the final score for image feature model 1 (IFM1; Fig. 4b). Here, CD8A is linked to colorectal carcinoma.