Fig. 2.

a Ki67-expresseing nuclei were stained by MIB-1 and appear as brown spots. Abnormal extreme dark spots caused errors in automated analyses and were excluded manually (see blue loops). b Dark blue regions designate cells in which the nuclei did not express Ki67, and yellow and orange regions designate cells with Ki67-expressing nuclei (note, orange signifies more intense expression). c CD34-stained vessels appear brown. d Detected vessels were highlighted in neon green using the Aperio algorithm. e The raw image was segmented into cancer (pink) versus non-cancerous (green) areas as shown in f. g Individual cells were detected as neon green loops. Based on nuclear shape, the compactness of the nuclei was calculated using inForm 1.0.2 (Perkin Elmer)