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. 2018 Aug 24;8:12815. doi: 10.1038/s41598-018-31153-7

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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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ACT process and contributions of lipid-extraction, RI-matching, and size expansion to tissue transparency (a) Schematic diagram of refractive index (RI) matching and the active CLARITY technique (ACT) process. (b) Images of 1-mm-thick tissue slices during the tissue-clearing process. Comparison of two different steps provides contributions of each factor (bottom) (c) Transparency of each organ tissue during the tissue-clearing process. (d) Size changes of each organ at each tissue-clearing step. (e) Contributions of RI-matching and lipid extraction to tissue clearing, if the effect of size change on transparency is compensated. (f) 3D plot of contributions of lipid-extraction and RI-matching to tissue transparency. The color indicates the final transparency (red-green-blue). N = 3 for each organ.