Exploring the human cerebral cortex using confocal microscopy

Anno: 2021

Autori: Pesce Luca; Laurino Annunziatina; Scardigli Marina; Yang Jiarui; Boas David A.; Hof Patrick R.; Destrieux Christophe; Costantini Irene; Pavone Francesco Saverio

Affiliazione autori: European Laboratory for Non-linear Spectroscopy (LENS), University of Florence, Sesto Fiorentino, FI, Italy; Department of Physics and Astronomy, University of Florence, Sesto Fiorentino, FI, Italy; Boston University, Department of Biomedical Engineering, Boston, USA; Nash Family Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA; UMR 1253, iBrain, Universita de Tours, Inserm, Tours, France; Department of Biology, University of Florence, Italy; National Institute of Optics (INO), National Research Council (CNR), Italy

Abstract: Cover-all mapping of the distribution of neurons in the human brain would have a significant impact on the deep understanding of brain function. Therefore, complete knowledge of the structural organization of different human brain regions at the cellular level would allow understanding their role in the functions of specific neural networks. Recent advances in tissue clearing techniques have allowed important advances towards this goal. These methods use specific chemicals capable of dissolving lipids, making the tissue completely transparent by homogenizing the refractive index. However, labeling and clearing human brain samples is still challenging. Here, we present an approach to perform the cellular mapping of the human cerebral cortex coupling immunostaining with SWITCH/TDE clearing and confocal microscopy. A specific evaluation of the contributions of the autofluorescence signals generated from the tissue fixation is provided as well as an assessment of lipofuscin pigments interference. Our evaluation demonstrates the possibility of obtaining an efficient clearing and labeling process of parts of adult human brain slices, making it an excellent method for morphological classification and antibody validation of neuronal and non-neuronal markers. (C) 2021 The Authors. Published by Elsevier Ltd.

Giornale/Rivista: PROGRESS IN BIOPHYSICS & MOLECULAR BIOLOGY

Volume: 168      Da Pagina: 3  A: 9

Maggiori informazioni: 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. We express gratitude to the two donors involved in the body donation programs of the Association des dons du corps du Centre Ouest, Tours, and of the Massachusetts General Hospital. They made this study possible through their generous donation to science. This project has received funding from the European Union’s Horizon 2020 research and innovation Framework Programme under grant agreement No. 654148 (LaserlabEurope), from the 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), from the General Hospital Corporation Center of the National Institutes of Health under award number U01 MH117023, and from the Italian Ministry for Education in the framework of Euro-Bioimaging Italian Node (ESFRI research infrastructure). Finally, this research was carried out with the contribution from 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.
Parole chiavi: Human brain; Clearing; SWITCH2
DOI: 10.1016/j.pbiomolbio.2021.09.001

Citazioni: 17
dati da “WEB OF SCIENCE” (of Thomson Reuters) aggiornati al: 2026-02-01
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