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
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.
PLoS One. 2016 Jun 30;11(6):e0158628. doi: 10.1371/journal.pone.0158628. eCollection 2016.
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.
通过使组织透明化，现在已经能够实现完整组织的三维可视化。CLARITY是一种独特的组织透明化技术，其特点是使用去污剂从固定组织中去除脂质以实现光学透明。为了保持组织完整性，有人提出用基于丙烯酰胺的水凝胶包埋组织。在本研究中，我们研究了在CLARITY中使用丙烯酰胺的原理，并提供证据表明在CLARITY中省略丙烯酰胺 - 水凝胶包埋不会改变固定组织中组织形态和分子信息的保存。因此，我们提出了一种用于甲醛固定组织透明化的新颖且简化的工作流程，这将有助于该技术在实验室中的应用。此外，我们详细研究了基本的组织透明化过程，并强调了一些针对三维组织学这一新兴学科所必需的技术进行有针对性改进的领域。
