PUBLICATION
Optimised tissue clearing minimises distortion and destruction during tissue delipidation
- Authors
- Lee, K., Lai, H.M., Soerensen, M.H., Hui, E.S., Ma, V.W., Cho, W.C., Ho, Y.S., Chang, R.C.
- ID
- ZDB-PUB-211123-24
- Date
- 2021
- Source
- Neuropathology and applied neurobiology 47: 441-453 (Journal)
- Registered Authors
- Keywords
- antigenicity, delipidation, opacity, protein loss, three-dimensional imaging, transparent brain
- MeSH Terms
-
- Surface-Active Agents
- Animals
- Zebrafish
- Rats, Sprague-Dawley
- Imaging, Three-Dimensional/methods*
- Artifacts
- Microscopy, Confocal/methods
- Sodium Dodecyl Sulfate
- Rats
- Brain*
- Female
- Male
- Mice
- Histocytological Preparation Techniques/methods*
- PubMed
- 33107057 Full text @ Neuropathol. Appl. Neurobiol.
Citation
Lee, K., Lai, H.M., Soerensen, M.H., Hui, E.S., Ma, V.W., Cho, W.C., Ho, Y.S., Chang, R.C. (2021) Optimised tissue clearing minimises distortion and destruction during tissue delipidation. Neuropathology and applied neurobiology. 47:441-453.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping