This protocol is a detailed version of the SHORT clearing and labeling method optimized by the European Laboratory for Non-Linear Spectroscopy - University of Florence -Italy, which describes the process to optically clear and fluorescently label human brain volumetric slabs, permitting the 3D reconstruction of the tissue at micrometer-resolution using light-sheet fluorescence microscopy (LSFM).

Note: Research reported in this publication was supported by the European Union's Horizon 2020 research and innovation Framework Programme under grant agreement No. 654148 (Laserlab-Europe); European Union's Horizon 2020 Framework Programme for Research and Innovation under the Specific Grant Agreement No. 785907 (Human Brain Project SGA2) and No. 945539 (Human Brain Project SGA3); Italian Ministry for Education in the framework of Euro-Bioimaging Italian Node (ESFRI research infrastructure); "Fondazione CR Firenze" (private foundation) and the BRAIN Initiative Cell Census Network of the National Institutes of Health under award number U01 MH117023. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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