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. 2018 Jun 21;8:9431. doi: 10.1038/s41598-018-27504-z

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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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Ex vivo human excisional cutaneous wound model of biofilm formation. Representative images of control, SA113, SA113ΔtagO, MSSA, PA and SP biofilms after 1, 3 and 5 days incubation. (A) Stereo-fluorescence aerial microscopy with LIVE/DEAD® (SYTO® 9 and PI) staining. Viable cells are stained green and non-viable cells are stained red. Scale bar: 250 µm. Wide-field fluorescence microscopy of cross-sections stained with DAPI + Con-A (B) and SYTO® 9 + Con-A (C). Viable bacterial cells are stained blue (DAPI) or green (SYTO® 9) and ECM is stained red (Con-A). White dashed line (B,C) indicates biofilm/wound tissue boundary. Scale bar: 10 µm. Scanning electron microscopy (SEM) images of MSSA (D), PA (E) and SP (F) biofilms. SA113, Staphylococcus aureus wild type strain; SA113ΔtagO, biofilm-forming deficient mutant derived from SA113; MSSA, methicillin sensitive Staphylococcus aureus ATCC 29213; PA, Pseudomonas aeruginosa; SP, Streptococcus pyogenes; PI, propidium iodide; DAPI, 4′,6-diamidino-2-phenylindole; Con-A, Concanavalin A.