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. 2020 Apr 3;10:5918. doi: 10.1038/s41598-020-62799-x

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© The Author(s) 2020

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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Collagen fiber organization within the wall of a pressurized, unfixed rat tail lymphatic vessel in tubular and valvular regions including reconstruction of SHG imaging of collagen fibers along with dorsal view ventral views (Panel A), the z-stack images obtained from SHG imaging of collagen fibers and respective fiber extractions using CT-FIRE algorithm (Panel B), distribution of collagen fiber angles (Panel C), distribution of collagen fiber straightness within the vessel wall for low (~2 mmHg) and high (~8 mmHg) pressure scenarios (Panel D), and the average distribution of collagen fibers (n = 5) plotted as the mean value along with the standard deviation (Panel E, the width of each bin is 20° and values of x-axis indicate the center of each bin). The vessels were excised from the base of the tail and mounted on glass pipettes in a custom-made vessel chamber compatible with the microscope. The vessels were unfixed, the pressure was ~2 mmHg, and the axial stretch was ~10%. The axial direction corresponds to 0° and the circumferential direction corresponds to ±90°.