A Comparative Performance Analysis of Multispectral and RGB Imaging on HER2 Status Evaluation for the Prediction of Breast Cancer Prognosis Wenlou Liu,

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A Comparative Performance Analysis of Multispectral and RGB Imaging on HER2 Status Evaluation for the Prediction of Breast Cancer Prognosis Wenlou Liu, Linwei Wang, Jiuyang Liu, Jingping Yuan, Jiamei Chen, Han Wu, Qingming Xiang, Guifang Yang, Yan Li Translational Oncology Volume 9, Issue 6, Pages 521-530 (December 2016) DOI: 10.1016/j.tranon.2016.09.007 Copyright © 2016 The Authors Terms and Conditions

Figure 1 Flowchart of study design and major technical procedures. (A1) TMAs with 480 cores of 240 BC specimens were constructed. (A2) Three key molecule biomarkers of BC were stained by routine IHC method, staining HER2 in cell membrane. (B1–B3) The HER2 MS and RGB images were captured by MSI system, and their unmixing and quantification were obtained by CRi Nuance software. (C1–C3) HER2 MS and RGB image analysis outputs were analyzed by X-tile software (C1). All patients were divided into two groups (C2) and analyzed by Kaplan-Meier method (C3). Translational Oncology 2016 9, 521-530DOI: (10.1016/j.tranon.2016.09.007) Copyright © 2016 The Authors Terms and Conditions

Figure 2 Immunohistological findings in BC TMAs. (A–C) Representative examples of HER2 scored as 1+, 2+, and 3+. (D–F) The RGB images were acquired by traditional RGB imaging system. (G–I) The corresponding MS images were acquired by MSI system. (D and G) Positive HER2 membrane staining was scored as 1+. (E and H) Positive HER2 membrane staining was scored as 2+. (F and I) Positive HER2 membrane staining was scored as 3+. Magnification 40× (A, B, and C) or 100× (D, E, F, G, H, and I); scale bar: 100 μm for all images. Translational Oncology 2016 9, 521-530DOI: (10.1016/j.tranon.2016.09.007) Copyright © 2016 The Authors Terms and Conditions

Figure 3 The relationship between total signal OD index and 5-DFS in BC patients, which was obtained from the analysis of HER2 MS or HER2 RGB images. HER2 MS (A) and HER2 RGB (B) are negatively related with 5-DFS in all patients. HER2 MS (C) and HER2 RGB (D) are inversely related with 5-DFS in LN-positive patients. HER2 MS (E) is negatively related with 5-DFS in LN-negative patients, but HER2 RGB (F) has no statistical significance in LN-negative patients. P value tested by the log-rank test. Translational Oncology 2016 9, 521-530DOI: (10.1016/j.tranon.2016.09.007) Copyright © 2016 The Authors Terms and Conditions

Figure 4 The relationship between average signal OD index and 5-DFS in BC patients, which was obtained from the analysis of HER2 MS or HER2 RGB images. (A–F) Similar results were further observed. P value tested by the log-rank test. Translational Oncology 2016 9, 521-530DOI: (10.1016/j.tranon.2016.09.007) Copyright © 2016 The Authors Terms and Conditions

Figure 5 ROC curve analysis on the prognostic value of HER2 MS and HER2 RGB. (A) By the analysis of total signal OD index, AUC of HER2 MS was greater than HER2 RGB and clinical stage but smaller than LN status and histological grade on recurrence. (B) Similar results were further realized by the determination of average signal OD index. Translational Oncology 2016 9, 521-530DOI: (10.1016/j.tranon.2016.09.007) Copyright © 2016 The Authors Terms and Conditions