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. 2017 Aug 29;7:9895. doi: 10.1038/s41598-017-10204-5

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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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Comparison of the three-dimensional structures of individual microglia. (A–G) Selected microglia from the topmost scanned layers (Z = 0–50 μm) of each XYZ acquisition from the different clearing protocols. The finest and most visibly continuous branches are achieved with the FACT protocol. (H) Comparisons of microglial branch length among the seven clearing protocols using the semiautomatic Imaris Filament algorithm. The highest signals were detected with the FACT and CLARITY - 37 °C protocols. All confocal images were captured at 25× magnification. Scale bars are 20 μm.