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. 2019 Feb 28;9:3148. doi: 10.1038/s41598-019-39760-8

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© The Author(s) 2019

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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Reliability of values compared to manual ImageJ-based evaluation. (a,b) The manual analysis of a narrow area over a single nucleus using the ImageJ Plot Profile function varies considerably from the plot generated from a whole nucleus frame. The scale bar represents 5 µm. (c) The Stripenator analyses a variety of values of each damage area (1–4) including area size, mean and maximal intensity. These values are measured independently of the size, angle and position of the damage areas. The images show the area of a single nucleus with a γH2A.X DNA damage reference staining. The scale bar represents 5 µm. (d) Analysis of evaluation time was performed by two users with three different methods. User-variability was measured as difference of values between two users using the three different analysing methods (e) and between the method wand (tracing) tool and the Stripenator for an untrained user and an expert (f).