title: Rationalisation and Validation of an Acrylamide-Free Procedure in Three-Dimensional Histological Imaging.
abstract: Three-dimensional visualization of intact tissues is now being achieved by turning tissues transparent. CLARITY is a unique tissue clearing technique, which features the use of detergents to remove lipids from fixed tissues to achieve optical transparency. To preserve tissue integrity, an acrylamide-based hydrogel has been proposed to embed the tissue. In this study, we examined the rationale behind the use of acrylamide in CLARITY, and presented evidence to suggest that the omission of acrylamide-hydrogel embedding in CLARITY does not alter the preservation of tissue morphology and molecular information in fixed tissues. We therefore propose a novel and simplified workflow for formaldehyde-fixed tissue clearing, which will facilitate the laboratory implementation of this technique. Furthermore, we have investigated the basic tissue clearing process in detail and have highlighted some areas for targeted improvement of technologies essential for the emerging subject of three-dimensional histology.
authors.affiliation: School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.|Neuropathology Unit, Division of Brain Sciences, Department of Medicine, Imperial College London, United Kingdom.
authors.forename: Hei Ming
authors.initials: HM
authors.lastname: Lai
authors.affiliation: School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.|Neuropathology Unit, Division of Brain Sciences, Department of Medicine, Imperial College London, United Kingdom.
authors.forename: Alan King Lun
authors.initials: AK
authors.lastname: Liu
authors.affiliation: Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom.
authors.forename: Wai-Lung
authors.initials: WL
authors.lastname: Ng
authors.affiliation: Neuropathology Unit, Division of Brain Sciences, Department of Medicine, Imperial College London, United Kingdom.
authors.forename: John
authors.initials: J
authors.lastname: DeFelice
authors.affiliation: Electron Microscope Unit, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
authors.forename: Wing Sang
authors.initials: WS
authors.lastname: Lee
authors.affiliation: School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
authors.forename: Heng
authors.initials: H
authors.lastname: Li
authors.affiliation: School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Li
authors.affiliation: School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
authors.forename: Ho Man
authors.initials: HM
authors.lastname: Ng
authors.affiliation: School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.|State Key Laboratory of Brain and Cognitive Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.|Research Centre of Heart, Brain, Hormone and Healthy Aging, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
authors.forename: Raymond Chuen-Chung
authors.initials: RC
authors.lastname: Chang
authors.affiliation: School of Optometry, The Hong Kong Polytechnic University, Hung Hom, Hong Kong SAR, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Lin
authors.affiliation: School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.|State Key Laboratory of Brain and Cognitive Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.|Research Centre of Heart, Brain, Hormone and Healthy Aging, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.|Joint Laboratory of Jinan University and The University of Hong Kong, GHM Institute of CNS Regeneration, Jinan University, Guangzhou, China.
authors.forename: Wutian
authors.initials: W
authors.lastname: Wu
authors.affiliation: Neuropathology Unit, Division of Brain Sciences, Department of Medicine, Imperial College London, United Kingdom.
authors.forename: Steve M
authors.initials: SM
authors.lastname: Gentleman
mesh_terms: D020106:Acrylamide; D000818:Animals; D001921:Brain / Q000033:anatomy & histology*; D002853:Chromatography, Liquid; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D016707:Tissue Fixation / Q000379:methods*
chemical_list: D020106:Acrylamide