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. 2016 Jan 14;16:5. doi: 10.1186/s12880-016-0105-4

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© Fliedner et al. 2016

Open AccessThis 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. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 6 — Immunohistochemistry (IHC) sections of micro vessel density (CD31 staining), proliferating cells (Ki-67 staining), and non-viable cell count (HE staining). Selection of IHC results presented to illustrate collected tissue samples and analyzed tissue. a CD31 staining visualizing endothelial cells in vessels of tumor slide. Staining made with antibody concentration of 1:50. Picture originates from mouse in the −MG group. Arrows indicate positive staining of vessels in slide. b Edited version of picture A after using the CAIMAN segmentation algorithm to detect antibody stained areas. Arrows indicate vessels identified by the CAIMAN algorithm (marked by thin green contouring line). c Ki-67 staining for proliferating cells in tumor section from a mouse in the +MG group. Staining made using antibody concentration of 1:400. d Review of image C after using the ImmunoRatio application for selecting proliferating cell percentage. Cells marked in brown are considered as positive Ki-67 and blue cells negative. e Haematoxylin eosin staining for non-viable cell count in tumor slide from mouse in the +MG group. f Image E after separation of viable cells (red), non-viable cells (green), and background (purple) with the Advanced Weka Segmentation tool in FiJi (ImageJ)