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

Hei Ming Lai, Alan King Lun Liu, Harry Ho Man Ng, Marc H. Goldfinger, Tsz Wing Chau, John DeFelice, Bension S. Tilley, Wai Man Wong, Wutian Wu () and Steve M. Gentleman ()
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Hei Ming Lai: The University of Hong Kong
Alan King Lun Liu: The University of Hong Kong
Harry Ho Man Ng: The University of Hong Kong
Marc H. Goldfinger: Imperial College London, Burlington Danes Building, Hammersmith Campus
Tsz Wing Chau: Imperial College London, Burlington Danes Building, Hammersmith Campus
John DeFelice: Imperial College London, Burlington Danes Building, Hammersmith Campus
Bension S. Tilley: Imperial College London, Burlington Danes Building, Hammersmith Campus
Wai Man Wong: The University of Hong Kong
Wutian Wu: The University of Hong Kong
Steve M. Gentleman: Imperial College London, Burlington Danes Building, Hammersmith Campus

Nature Communications, 2018, vol. 9, issue 1, 1-12

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-03359-w

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