Rationalisation and validation of an acrylamide-free procedure in three-dimensional histological imaging

Hei Ming Lai, Alan King Lun Liu, Wai Lung Ng, John DeFelice, Wing Sang Lee, Heng Li, Wen Li, Ho Man Ng, Raymond Chuen Chung Chang, Bin Lin, Wutian Wu, Steve M. Gentleman

Research output: Journal article publicationJournal articleAcademic researchpeer-review

30 Citations (Scopus)


This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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 CLAR-ITY 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 threedimensional histology.
Original languageEnglish
Article numbere0158628
JournalPLoS ONE
Issue number6
Publication statusPublished - 1 Jun 2016

ASJC Scopus subject areas

  • General Biochemistry,Genetics and Molecular Biology
  • General Agricultural and Biological Sciences


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