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. 2018 Jan 8;8:5. doi: 10.1038/s41598-017-18501-9

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© The Author(s) 2017

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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Live-imaging quantification of P. aeruginosa cell infection by fluorescence intensity measurement of Hoechst-labelled nuclei. HUVEC, CHO, 3T3, HeLa and A549 cells were labelled with vital-Hoechst prior to infection with P. aeruginosa and monitored by microscopy at a 5x magnification. (a) Cell nuclei observed by fluorescent labelling and automatically segmented (insert). Nuclei with intensities below a fixed threshold were delineated in magenta while those above the threshold were delineated in green. The scale bar shown on the first image of the HUVECs cells corresponds to 200 µm. (b) Kinetic plots representing the percentage of nuclei with intensities above the thresholds in the images taken every 15 min.