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. 2017 Jun 13;7:3419. doi: 10.1038/s41598-017-03548-5

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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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THG reveals interfaces in cortical bone. (a) THG image of a transverse section of a bovine femur at a depth of 10 µm within the sample, revealing the LCN (lacunae indicated with arrowheads) and the cement line of the osteons (stars). Circle, Haversian channel. The white arrow shows a canalicule crossing the cement line. Scale bar, 50 µm. (b) Zoom on the white box shown in (a), revealing the two interfaces of the canaliculi. Scale bar, 10 µm. (c) Profile along the red box in (a), illustrating the reduced signal at the cement line (arrow). (d) Profile across a canalicule along the green line in b providing an estimate for the canalicule diameter D and for the lateral resolution. (e) Distribution of canalicule diameters D estimated from THG images in the volume shown in Supplementary Movie 1 including image (b).