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In Situ Ca2+ Imaging of the Enteric Nervous System
Published on: January 29, 2015
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Expansion Microscopy of the Enteric Nervous System: A Feasibility Study.
Xin Xu1,2,3, Wenchuan Zhang3, Menachem Hanani1,2
1Laboratory of Experimental Surgery, Hadassah-Hebrew University Medical Center, Mount Scopus, Jerusalem 91240, Israel.
Cells
|September 26, 2025
Summary
We developed a new protocol for expansion microscopy (ExM) to visualize the enteric nervous system (ENS) in mice. This method enhances resolution, allowing detailed study of ENS structures with standard microscopes.
Area of Science:
- Neuroscience
- Microscopy techniques
Background:
- Expansion microscopy (ExM) offers nanoscale resolution for light microscopy by physically enlarging specimens.
- ExM is widely used in neurobiology but not yet adapted for the enteric nervous system (ENS).
Purpose of the Study:
- To provide a detailed and reproducible protocol for applying ExM to mouse colonic ENS tissue.
- To enable high-resolution structural analysis of the ENS.
Main Methods:
- The protocol involves tissue preparation, histochemical and immunostaining (NADPH-diaphorase, GFAP), biomolecule anchoring, gelation, proteinase K digestion, and isotropic expansion.
- Step-by-step instructions, reagents, and critical parameters are detailed for robustness.
Main Results:
- The protocol achieves 3-5 fold tissue expansion, enabling clear visualization of neuronal and glial structures.
- Tissue architecture is preserved with minimal distortion (approx. 7% in X-Y plane).
Conclusions:
- This protocol offers a reliable method for high-resolution structural analysis of the ENS.
- The method can be adapted for other peripheral tissues.
