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. 2018 Feb 21;8:3418. doi: 10.1038/s41598-018-21757-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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A custom algorithm was created to identify and quantitate PCK^+ve cells in immunohistochemically stained samples. (A) PCK^+ve and PCK^−ve cells were distinguished based on DAB levels. (B) Representative images of PCK^+ve cells (insert 1) and PCK^−ve cells (insert 2) are shown. (C) InForm output had a high correlation with manual counting (r² = 0.9202, p < 0.0001, n = 14 FOVs). (D) Comparative images of cells counted manually (red indicates PCK^+ve cells, green indicates nuclei of cells that are PCK^−ve), and with InForm (green indicates all nuclei counted by InForm, red indicates PCK^+ve nuclei). Scale bar 200 μm. ***p < 0.0001.