Figure 3.

Scoring the stained proteins. A. Pipeline for tissues and cells segment by InForm software. The image of raw multiple fluorescent channels merged was separated into two fluorescent channels (Opal670 and DAPI) (upper). Segment tissue was divided into cancer and stroma based on the fluorescent signal intensity of DAPI (middle panel). Representative image of 0–3+(4-bin) scoring system (bottom). This score system can be used to calculate H-score with cell stains. Difference colors displays four levels (0∼1, 1∼2, 2∼3, 3∼) and percentage positivity of cells with each bin of stained protein. H-score was automatic calculated by InForm software. B. Unsupervised hierarchical clustering of eight TGF-β signaling proteins classified the 90 patients with CRC.