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. 2021 Apr 13;11:8038. doi: 10.1038/s41598-021-86092-7

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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The MAGIC protocol. (a) Representative TPFM images of a mouse brain section during the three subsequent steps of the MAGIC protocol: fixation (PFA), glycerolization (Gly), and washing (MAGIC). The images show different areas of the caudate putamen. For each condition, a fiber bundle is indicated with a red arrow. Scale bar = 50 μm. (b) 3D reconstruction of myelinated fibers, imaging performed with TPFM at the resolution of (0.44 × 0.44 × 1) μm³. Box scale = 10 μm. (c) Measurement (mean ± std.err) of photons emitted by the myelinated fibers (fiber) and the surrounding tissue (tissue) during the different steps of the protocol (PFA, Gly, MAGIC) detected at different excitation wavelengths. (d) Intensity contrast (mean ± std.err) observed at different excitation wavelengths during the three steps of the MAGIC protocol. (e) Images of a mouse brain section treated with MAGIC and labeled with FluoroMyelin red. In green and red respectively, the autofluorescence and the exogenous signals are shown. Images were obtained with TPFM, scale bar = 50 μm. Images and the 3D rendering were prepared using Fiji (www.fiji.sc/Fiji) ²⁰, graphs were prepared using OriginPro 9.0 (www.originlab.com).