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Nat Commun. 2018 Mar 14;9(1):1066. doi: 10.1038/s41467-018-03359-w.

Next generation histology methods for three-dimensional imaging of fresh and archival human brain tissues.

Author information

1
School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, 852, Hong Kong.
2
Neuropathology Unit, Division of Brain Sciences, Imperial College London, Burlington Danes Building, Hammersmith Campus, Du Cane Road, London, W12 0NN, UK.
3
School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, 852, Hong Kong. wtwu@hku.hk.
4
GMH Institute of CNS Regeneration, Jinan University, 601 Huangpu Avenue West, Guangzhou, 510632, China. wtwu@hku.hk.
5
Re-Stem Biotechnology Co., Ltd, 2588 Wuzhong Road, Suzhou, 215310, China. wtwu@hku.hk.
6
Neuropathology Unit, Division of Brain Sciences, Imperial College London, Burlington Danes Building, Hammersmith Campus, Du Cane Road, London, W12 0NN, UK. s.gentleman@imperial.ac.uk.

Abstract

Modern clearing techniques for the three-dimensional (3D) visualisation of neural tissue microstructure have been very effective when used on rodent brain but very few studies have utilised them on human brain material, mainly due to the inherent difficulties in processing post-mortem tissue. Here we develop a tissue clearing solution, OPTIClear, optimised for fresh and archival human brain tissue, including formalin-fixed paraffin-embedded material. In light of practical challenges with immunostaining in tissue clearing, we adapt the use of cresyl violet for visualisation of neurons in cleared tissue, with the potential for 3D quantification in regions of interest. Furthermore, we use lipophilic tracers for tracing of neuronal processes in post-mortem tissue, enabling the study of the morphology of human dendritic spines in 3D. The development of these different strategies for human tissue clearing has wide applicability and, we hope, will provide a baseline for further technique development.

PMID:
29540691
PMCID:
PMC5852003
DOI:
10.1038/s41467-018-03359-w
[Indexed for MEDLINE]
Free PMC Article

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