Exploring the human cerebral cortex using confocal microscopy

Year: 2021

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

Autors Affiliation: 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.


Volume: 168      Pages from: 3  to: 9

KeyWords: Human brain; Clearing; SWITCH2
DOI: 10.1016/j.pbiomolbio.2021.09.001

Citations: 9
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