3D molecular phenotyping of cleared human brain tissues with light-sheet fluorescence microscopy
Year: 2022
Authors: Pesce Luca; Scardigli Marina; Gavryusev Vladislav; Laurino Annunziatina; Mazzamuto Giacomo; Brady Niamh; Sancataldo Giuseppe; Silvestri Ludovico; Destrieux Christophe; Hof Patrick R.; Costantini Irene; Pavone Francesco S.
Autors Affiliation: Univ Florence, European Lab Nonlinear Spect LENS, Sesto Fiorentino, Italy; Univ Florence, Dept Phys & Astron, Sesto Fiorentino, Italy; Natl Res Council CNR, Natl Inst Opt INO, Sesto Fiorentino, Italy; Univ Tours, iBrain, UMR 1253, Tours, France; Icahn Sch Med Mt Sinai, Nash Family Dept Neurosci, New York, NY 10029 USA; Icahn Sch Med Mt Sinai, Friedman Brain Inst, New York, NY 10029 USA; Univ Florence, Dept Biol, Florence, Italy; Univ Florence, Dept Neurosci Psychol Drug Res & Child Hlth NEURO, Florence, Italy.
Abstract: The combination of optical tissue transparency with immunofluorescence allows the molecular characterization of biological tissues in 3D. However, adult human organs are particularly challenging to become transparent because of the autofluorescence contributions of aged tissues. To meet this challenge, we optimized SHORT (SWITCH–HO–antigen Retrieval–TDE), a procedure based on standard histological treatments in combination with a refined clearing procedure to clear and label portions of the human brain. 3D histological characterization with multiple molecules is performed on cleared samples with a combination of multi-colors and multi-rounds labeling. By performing fast 3D imaging of the samples with a custom-made inverted light-sheet fluorescence microscope (LSFM), we reveal fine details of intact human brain slabs at subcellular resolution. Overall, we proposed a scalable and versatile technology that in combination with LSFM allows mapping the cellular and molecular architecture of the human brain, paving the way to reconstruct the entire organ.
Journal/Review: COMMUNICATIONS BIOLOGY
Volume: 5 (1) Pages from: 447-1 to: 447-12
More Information: We thank Leah Morgan and Bruce Fischl, Massachusetts General Hospital, A.A. Martinos Center for Biomedical Imaging, Department of Radiology, USA for providing the human brain specimen 2 analyzed in this study and Jiarui Yang and David Boas, Boston University, Department of Biomedical Engineering, USA, for performing the slicing of the sample. We express our gratitude to the donor involved in the body donation program of the Association des dons du corps du Centre Ouest, Tours, and of the Massachusetts General Hospital who made this study possible by generously donating his body to science. This project has received funding from: European Unionīs Horizon 2020 research and innovation Framework Programme under grant agreement No. 654148 (Laserlab-Europe); European Unionīs Horizon 2020 Framework Programme for Research and Innovation under the Specific Grant Agreement No. 785907 (Human Brain Project SGA2), No. 945539 (Human Brain Project SGA3) and under the Marie Skodowska-Curie grant agreement No. 793849 (MesoBrainMicr); General Hospital Corporation Center of the National Institutes of Health under award number U01 MH117023; Italian Ministry for Education in the framework of Euro-Bioimaging Italian Node (ESFRI research infrastructure); “Fondazione CR Firenze” (private foundation). The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.KeyWords: light-sheet microscopy, human brain tissueDOI: 10.1038/s42003-022-03390-0ImpactFactor: 5.900Citations: 21data from “WEB OF SCIENCE” (of Thomson Reuters) are update at: 2024-11-03References taken from IsiWeb of Knowledge: (subscribers only)Connecting to view paper tab on IsiWeb: Click hereConnecting to view citations from IsiWeb: Click here
