Quantitative assessment of optical clearing methods on formalin-fixed human lymphoid tissue
Yvonne Schega1, Nadine Flinner2, Martin-Leo Hansmann3
1University Clinic Frankfurt am Main, Department of Pathology, Goethe University, Germany.
Pathology, Research and Practice
|August 22, 2020
Summary
Optical clearing methods enable high-resolution 3D imaging of tissues. Researchers evaluated eight methods on human tonsils, finding Ce3D effective and less toxic for clearing lymphoid tissue.
Area of Science:
- Biomedical Imaging
- Histology
- Optical Engineering
Background:
- Three-dimensional (3D) volume imaging requires effective optical clearing of biological tissues for cellular-level resolution.
- Existing clearing protocols primarily focus on murine central nervous system tissue, with limited investigation into peripheral human organs.
Purpose of the Study:
- To evaluate and compare eight established optical clearing methods for formaldehyde-fixed human tonsils.
- To provide an application-oriented taxonomy of clearing techniques for lymphatic tissues, considering efficacy, speed, fluorescence preservation, toxicity, and cost.
Main Methods:
- Tested eight optical clearing methods: BABB, Ce3D, CUBIC, ECi, ChemScale, ChemScaleQQ5, SeeDB2, and PACT.
- Applied methods to formaldehyde-fixed human tonsil samples.
- Assessed transparency, clearing speed, fluorescence preservation, toxicity, preparation time, and cost.
Main Results:
- All tested clearing protocols successfully rendered human tonsil tissues transparent.
- Ce3D and PACT demonstrated the highest degrees of tissue transparency.
- Ce3D exhibited lower toxicity and required less preparation and processing time compared to other methods.
Conclusions:
- Ce3D is recommended as an effective and practical optical clearing method for human lymphoid tissue.
- The study provides valuable data to guide researchers in selecting appropriate clearing techniques for lymphatic tissues.
