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Whole-mouse clearing and imaging at the cellular level with vDISCO
Ruiyao Cai1,2, Zeynep Ilgin Kolabas1,2,3, Chenchen Pan1,2
1Institute for Tissue Engineering and Regenerative Medicine (iTERM), Helmholtz Munich, Munich, Germany.
Nature Protocols
|January 25, 2023
Summary
We developed nanobody-boosted 3D imaging of solvent-cleared organs (vDISCO) to make whole mice transparent for cellular-resolution imaging. This method enhances and stabilizes fluorescence, enabling comprehensive whole-body analysis.
Area of Science:
- Biology
- Biotechnology
- Microscopy
Background:
- Homeostatic and pathological processes impact multiple organs systemically.
- Advances in tissue clearing and microscopy enable holistic biological sample examination.
Purpose of the Study:
- To present a detailed protocol for nanobody-boosted 3D imaging of solvent-cleared organs (vDISCO).
- To enable whole-body imaging at cellular resolution for comprehensive biological analysis.
Main Methods:
- vDISCO is a pressure-driven, nanobody-based whole-body immunolabeling and clearing technique.
- The protocol renders whole mice transparent within 3 weeks, enhancing and stabilizing fluorescent signals.
- The method is accessible to labs with limited tissue clearing expertise.
Main Results:
- vDISCO significantly boosts fluorescence signal from genetically expressed reporters.
- The method allows reliable detection and quantification of fluorescent signals in intact rodents.
- Applications include whole-mouse neuronal mapping, metastasis detection, lymphatic system visualization, virus tracing, and brain pericyte imaging.
Conclusions:
- vDISCO provides a versatile pipeline for systematic, comprehensive studies of cellular phenomena and connectivity in whole bodies.
- This technique facilitates the analysis of entire organisms at cellular resolution.
