Nov 16, 2022

Public workspaceSHORT: a human brain tissue clearing and labeling protocol for 3D recostruction with LSFM

  • Luca Pesce1,
  • Marina Scardigli1,
  • Vladislav Gavryusev1,
  • Annunziatina Laurino1,
  • Giacomo Mazzamuto1,
  • Giuseppe Sancataldo1,
  • Ludovico Silvestri1,
  • Christophe Destrieux2,
  • Patrick R. Hof3,
  • Irene Costantini4,
  • Francesco S. Pavone1
  • 1European Laboratory for Non-Linear Spectroscopy - University of Florence -Italy;
  • 2UMR 1253, iBrain, Université de Tours, France;
  • 3Nash Family Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA;
  • 4European Laboratory for Non Linear Spectroscopy (LENS)
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Protocol CitationLuca Pesce, Marina Scardigli, Vladislav Gavryusev, Annunziatina Laurino, Giacomo Mazzamuto, Giuseppe Sancataldo, Ludovico Silvestri, Christophe Destrieux, Patrick R. Hof, Irene Costantini, Francesco S. Pavone 2022. SHORT: a human brain tissue clearing and labeling protocol for 3D recostruction with LSFM. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygxp6nkl8j/v1
Manuscript citation:
Pesce, L., Scardigli, M., Gavryusev, V. et al. 3D molecular phenotyping of cleared human brain tissues with light-sheet fluorescence microscopy. Commun Biol 5, 447 (2022). https://doi.org/10.1038/s42003-022-03390-0
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 05, 2021
Last Modified: November 16, 2022
Protocol Integer ID: 53808
Keywords: clearing, microscopy, SHORT, human brain, light sheet microscopy, LSFM, staining, labeling, immunofluorescence
Funders Acknowledgement:
European Union's Horizon 2020 research and innovation Framework Programme
Grant ID: 654148
European Union's Horizon 2020 Framework Programme for Research and Innovation
Grant ID: 785907
European Union's Horizon 2020 Framework Programme for Research and Innovation
Grant ID: 945539
Italian Ministry for Education in the framework of Euro-Bioimaging Italian Node
Grant ID: ESFRI research infrastructure
BRAIN Initiative Cell Census Network of the National Institutes of Health
Grant ID: U01 MH117023
Abstract
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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