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Updated: Dec 17, 2025
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Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids
Published on: June 5, 2020
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Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids
Ravian L van Ineveld1, Hendrikus C R Ariese1, Ellen J Wehrens1
1Princess Máxima Center for Pediatric Oncology; Department of Cancer Research, Oncode Institute, Hubrecht Institute-KNAW Utrecht; Cancer Genomics Center (CGC).
Journal of Visualized Experiments : Jove
|June 23, 2020
Summary
This study presents a new 3-day protocol for high-resolution 3D imaging of organoids using optical clearing and confocal microscopy. This method allows detailed cellular analysis of various organoid types for developmental and disease research.
Area of Science:
- Biotechnology
- Cell Biology
- Microscopy
Background:
- Organoid technology enables in vitro study of organ development, function, and disease.
- Fluorescence microscopy is crucial for characterizing organoid cellular composition and tissue similarity.
Purpose of the Study:
- To present a comprehensive protocol for high-resolution 3D imaging of whole organoids.
- To enable detailed cellular analysis and single-cell quantification of markers within organoids.
Main Methods:
- Immunofluorescent labeling of organoids.
- Optical clearing using FUnGI agent.
- 3D imaging with confocal microscopy.
Main Results:
- A widely applicable 3-day protocol for imaging diverse organoids (airway, colon, kidney, liver, tumor, mammary).
- Enabled high-resolution 3D imaging of whole organoids.
- Facilitated single-cell quantification of markers.
Conclusions:
- The developed protocol provides a robust method for detailed 3D organoid imaging.
- This technique advances research in organoid development, function, and disease modeling.
