Optimised tissue clearing minimises distortion and destruction during tissue delipidation

Krit Lee, Hei Ming Lai, Maja Hoejvang Soerensen, Edward Sai Kam Hui, Victor Wan San Ma, William Chi Shing Cho, Yuen Shan Ho, Raymond Chuen Chung Chang (Corresponding Author)

Research output: Journal article publicationJournal articleAcademic researchpeer-review

Abstract

Aims: A variety of tissue clearing techniques have been developed to render intact tissue transparent. For thicker samples, additional partial tissue delipidation is required before immersion into the final refractive index (RI)-matching solution, which alone is often inadequate to achieve full tissue transparency. However, it is difficult to determine a sufficient degree of tissue delipidation, excess of which can result in tissue distortion and protein loss. Here, we aim to develop a clearing strategy that allows better monitoring and more precise determination of delipidation progress. Methods: We combined the detergent sodium dodecyl sulphate (SDS) with OPTIClear, a RI-matching solution, to form a strategy termed Accurate delipidation with Optimal Clearing (Accu-OptiClearing). Accu-OptiClearing allows for a better preview of the final tissue transparency achieved when immersed in OPTIClear alone just before imaging. We assessed for the changes in clearing rate, protein loss, degree of tissue distortion, and preservation of antigens. Results: Partial delipidation using Accu-OptiClearing accelerated tissue clearing and better preserved tissue structure and antigens than delipidation with SDS alone. Despite achieving similar transparency in the final OPTIClear solution, more lipids were retained in samples cleared with Accu-OptiClearing compared to SDS. Conclusions: Combining the RI-matching solution OPTIClear with detergents, Accu-OptiClearing, can avoid excessive delipidation, leading to accelerated tissue clearing, less tissue damage and better preserved antigens.

Original languageEnglish
JournalNeuropathology and Applied Neurobiology
DOIs
Publication statusPublished - 26 Oct 2020

Keywords

  • antigenicity
  • delipidation
  • opacity
  • protein loss
  • three-dimensional imaging
  • transparent brain

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Neurology
  • Clinical Neurology
  • Physiology (medical)

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