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. 2018 Nov 23;8:17282. doi: 10.1038/s41598-018-35660-5

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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 bi-modal OEP histogram separating background signals and foci. Representative histograms showing the logarithmic OEP (a,c) or the inverse logarithmic OEP (b,d) for objects detected in unirradiated HOMSF1 cells (a,b) or HOMSF1 cells fixed at 24 h post 1 Gy (c,d). Coloured histograms in panels a and b show the results of the manual rating from Fig. 2a (average of the 3 experimenters). Since objects with OEP ratings near the manually defined foci threshold of 5 are located around the minimum in both representations, background signals and foci can be separated. The black lines in panels a and b indicate the mean of different algorithms determining the starting point for manual threshold adjustment.