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Updated: Jul 12, 2025
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Staining and High-Resolution Imaging of Three-Dimensional Organoid and Spheroid Models
Published on: March 27, 2021
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Advancements in 3D spheroid imaging: Optimised cryosectioning and immunostaining techniques.
Claire Charlet-Faure1,2, Annemette Præstegaard Thulesen1, Adelina Rogowska-Wrzesinska1
1Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, Odense M DK-5230, Denmark.
Methodsx
|October 20, 2023
Summary
Researchers developed a new protocol to improve the embedding and cryosectioning of spheroids and organoids, enhancing imaging quality for tissue research. This method preserves delicate structures, enabling clearer visualization of these complex biological models.
Area of Science:
- Biotechnology
- Cell Biology
- Histology
Background:
- Spheroids and organoids are vital 3D models mimicking in vivo tissue.
- Current embedding and sectioning methods can damage these fragile structures.
- Improved techniques are needed for high-resolution imaging of organoids and spheroids.
Purpose of the Study:
- To present a modified protocol for embedding and cryosectioning spheroids and organoids.
- To minimize structural distortion and damage during sample preparation.
- To enhance the quality of imaging for 3D cell culture models.
Main Methods:
- Optimized embedding containers and protocols were utilized.
- Cryosectioning was performed at optimized temperatures.
- Heat-induced antigen retrieval was applied to enhance immunostaining.
Main Results:
- The modified protocol significantly reduced damage to spheroids and organoids.
- Heat-induced antigen retrieval increased immunostaining intensity without compromising integrity.
- Thinner cryosections were achieved, resulting in clearer and more detailed images.
Conclusions:
- The developed protocol effectively preserves spheroid and organoid integrity.
- This method enhances imaging clarity and detail for 3D biological models.
- The protocol is suitable for widespread adoption in spheroid and organoid research.
