Fig. 1

Image analysis workflow for immunohistochemical staining quantification. a-d Prostate cancer tissue microarrays were stained by immunohistochemistry (IHC). Various staining qualities are highlighted. e-h Genie Histology Pattern Recognition software (Aperio) subclassified tumor areas into malignant epithelium (dark blue), stroma (yellow), and glass (cyan). i-l Within malignant epithelial areas, cell-based digital image analysis separately quantified cytoplasmic and nuclear staining within malignant epithelium using the Cytoplasmic algorithm (Aperio). Cytoplasmic staining intensities are pseudocolored for negative cytoplasmic (yellow), weak cytoplasmic (orange), medium cytoplasmic (dark orange), and strong cytoplasmic (red) staining. Nuclear staining intensities are pseudocolored for negative nuclear (cyan), weak nuclear (light blue), medium nuclear (blue), and strong nuclear (dark blue) staining. m-p Within malignant epithelial areas, area-based digital image analysis quantified total malignant epithelial area staining using the Color Deconvolution algorithm (Aperio). Area-based staining intensities are pseudocolored for negative (blue), weak (yellow), medium (orange), and strong (red) staining. Scale bars represent 50 μm