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Updated: Aug 5, 2026
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Staining and High-Resolution Imaging of Three-Dimensional Organoid and Spheroid Models
Published on: March 27, 2021
Histological Processing of Organoids for Immunostaining
Lisa Brossard1, Victor Perreaux1, Simon Vales1
1Nantes Université, CHU Nantes, Inserm, TENS, The Enteric Nervous System in Gut and Brain Diseases, IMAD, Nantes, France.
Bio-Protocol
|July 28, 2026
Summary
This study presents a new protocol for histology on human organoids, overcoming challenges with small, fragile samples. The method ensures reliable fixation, embedding, and sectioning for detailed analysis.
Area of Science:
- Stem cell biology
- Histology and histopathology
- Biotechnology
Background:
- Organoids are 3D stem cell-derived structures mimicking native tissues.
- Histological analysis is crucial for organoid characterization but challenging due to sample fragility.
- Standard paraffin embedding methods are often unsuitable for organoids.
Purpose of the Study:
- To develop a robust and reproducible protocol for the histological analysis of human organoids.
- To enable high-quality fixation, paraffin embedding, and sectioning of fragile organoid samples.
- To support detailed structural and cellular analysis of organoids for research.
Main Methods:
- Direct fixation of organoids within their culture matrix.
- Inclusion of organoids in HistoGel to prevent loss during processing.
- Adaptation of the protocol for intestinal and cardiac organoids.
- Compatibility with hematoxylin-eosin (H&E) staining and multiplex immunofluorescence.
Main Results:
- Successful fixation, embedding, and sectioning of fragile human organoids.
- Prevention of organoid loss during paraffin embedding using HistoGel.
- High-quality histological and immunofluorescence analysis achieved.
- Demonstrated applicability to various organoid types and staining techniques.
Conclusions:
- The developed protocol provides a cost-effective and accessible method for preserving and analyzing organoids.
- This technique enables detailed structural analysis and supports long-term storage of organoid samples.
- The method facilitates reliable histological and immunofluorescence studies on organoids.
