Rationalisation and Validation of an Acrylamide-Free Procedure in Three-Dimensional Histological Imaging

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...

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Bibliographic Details
Published in:	PloS one 2016-06, Vol.11 (6), p.e0158628-e0158628
Main Authors:	Lai, Hei Ming, Liu, Alan King Lun, Ng, Wai-Lung, DeFelice, John, Lee, Wing Sang, Li, Heng, Li, Wen, Ng, Ho Man, Chang, Raymond Chuen-Chung, Lin, Bin, Wu, Wutian, Gentleman, Steve M
Format:	Article
Language:	eng
Subjects:	Acrylamide Amides Animal cognition Animals Biology and Life Sciences Brain - anatomy & histology Brain research Chromatography, Liquid Clarity Clearing Detergents Embedding Formaldehyde Histology Humans Hydrogels Immunoglobulins Laboratory animals Lipids Mass Spectrometry Medicine Medicine and Health Sciences Mice Neuropathology Physical Sciences Preservation Proteins Research and Analysis Methods Tissue Fixation - methods Tissues Transparency Workflow
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Summary:	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.
ISSN:	1932-6203 1932-6203