Fig 5.

HER2(+) and HER2(-) testing cohort for DDSI staining and IHC validation. (A) Representative color, fluorescence, DDSI, H&E, and HER2 targeted IHC images of MCF7-HER2 (HER2+) and MCF7 (HER2-) tumor and adipose tissue pairs following staining using the optimal staining condition (Probe pair B, 200 nM concentration, 1 min incubation time). All images are representative of data collected for n=6 tumor and adipose tissue pairs per tumor cell line. All untargeted and targeted channel images are background corrected, normalized by their exposure time and calibration drop intensity, and displayed on equivalent color scales. DDSI images are displayed with equivalent color scales. H&E and IHC images were acquired from serial sections of the same tissue face imaged in the whole DDSI specimen images. Scale bars = 5 mm. (B) Tumor and normal tissue pixel intensity histograms, (C) ROC curves, and AUC values for untargeted, targeted, and DDSI images corresponding to each cell line. Optimal points determined from ROC analysis are displayed on each ROC curve (blue marker) and as a vertical line on each pixel value histogram. The optimal point determined from the training cohort data is displayed on the MCF7-HER2 tumor specimen ROC curve to demonstrate the diagnostic reproducibility under optimal staining conditions (orange marker). H&E: Hematoxylin & Eosin; IHC: immunohistochemistry; ROC: receiver operator characteristic; AUC: area under curve; opt pt: optimal point.