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. 2015 Nov 5;37(4):5013–5024. doi: 10.1007/s13277-015-4327-9

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© The Author(s) 2015

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

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Fig. 1 — The design and major technical procedures of this study. a Tumor tissue section slides were prepared from 86 selected patients with invasive BC. b Section slides were stained with three biomarkers in routine IHC, staining HER2 in cell membrane, CK5/6 in cytoplasm, and ER in cell nucleus. c1–c5 The MS (c1) and RGB images (c4) were captured and its spectral curves (c2, c5) were obtained by CRi Nuance software. d1–d4 The positive regions in the MS images (d1, d2) and RGB images (d3, d4) were unmixed and quantified by CRi Nuance software, respectively, and the quantitative results of MS and RGB images were compared. BC: breast cancer; HER2: human epidermal growth factor receptor-2; CK5/6: cytokeratin5/6; ER: estrogen receptor