Enhanced Spatial Mapping of Mouse Gastric Muscle Layers Using a Modified Swiss Roll Technique
November 25th, 2025
This protocol outlines a whole-stomach immunostaining method modified from the "Swiss roll" technique originally developed for the murine intestine.
Our research focuses on improving how we prepare and analyze the mild stomach for imaging. We've adapted the Swiss Roll Technique originally developed for the murine intestine so that all major gastric regions, including the fundus, corpus, and antrum, can be embedded together in a single cryo section. This allows us to visualize the stomach structure and cellular composition side by side while maintaining accurate spatial orientation and high image quality.
The stomach is a complex organ with delicate tissues. Small mistakes can damage cells or distort the arrangement, which makes high-resolution imaging and immuno staining inconsistent. Our Swiss roll technique solves this by keeping all regions, including the duodenum and esophagus intact in a single cryo block, making experiments more reliable and reproducible.
To begin, use scissors to cut along the ventral mesentery of the duodenum and stomach up to the pin at the corpus. Using a transfer pipette filled with PBS, flush out any residual food from the gastric lumen. Gently stretch the open stomach to achieve a flat orientation.
Pin the tissue flat again after the brief fixation. Using scissors, trim the stomach along the dashed outline to create a uniform shape suitable for rolling. Place the trimmed gastric tissue on PBS-moistened filter paper.
Roll the tissue into a Swiss roll configuration and secure it by inserting a toothpick through the center. Once rolled, insert a 30-gauge needle to secure its shape. Then bend the needle at a 90 degree angle to stabilize the rolled stomach.
Fix the rolled tissue in 4%paraformaldehyde at four degrees Celsius for six hours. Then gently remove the stabilizing needle. Embed the tissue in optimal cutting temperature compound using disposable base molds.
Then freeze the mold on dry ice. Once fully frozen, store the mold at minus 80 degrees Celsius. The new method enabled inclusion of the fundus, corpus, antrum, a portion of the esophagus, and duodenum within a single murine gastric Swiss roll section.
Hematoxylin and eosin staining of the gastric Swiss roll revealed well-preserved architecture in the fundus, corpus, antrum, esophagus, and duodenum with clearly visible mucosal, submucosal, and muscularis layers. Immunofluorescent staining showed expression of KIT and ANO1 in interstitial cells of Cajal and expression of beta tubulin three in enteric neurons, confirming cellular preservation in the gastric Swiss roll. KIT expression was also detected in paneth and goblet cells in the gastric mucosa while ANO1 was highly expressed in fundic columnar epithelial cells.
Beta tubulin three expression was observed in gastric, endocrine, and mucus secreting cells in addition to enteric neurons. Myosin heavy chain 11 staining confirmed the presence of smooth muscle cells and myofibroblasts in the gastric mucosa, while E-cadherin staining showed epithelial cells and KIT staining again marked interstitial cells of Cajal. The Swiss road technique can advance research on region-specific changes in gastric tissue.
Unlike traditional approaches that embed region separately, the Swiss Roll Technique processes all gastric regions under identical conditions, saving both time and reagents. This makes it a practical and scalable platform for comprehensive analysis of gastric tissue in both health and disease.
Overview
This protocol outlines a whole-stomach immunostaining method modified from the "Swiss roll" technique originally developed for the murine intestine. The method allows for the visualization of gastric structure and cellular composition while maintaining spatial orientation and image quality.
Key Study Components
Area of Science
- Neuroscience
- Gastroenterology
- Histology
Background
- The stomach is a complex organ with delicate tissues.
- Traditional methods often lead to inconsistencies in imaging and staining.
- The Swiss Roll Technique allows for embedding multiple gastric regions together.
- This enhances the reliability and reproducibility of experiments.
Purpose of Study
- To improve the preparation and analysis of gastric tissues for imaging.
- To visualize all major gastric regions in a single cryo section.
- To maintain accurate spatial orientation during imaging.
Methods Used
- Adaptation of the Swiss Roll Technique for whole-stomach immunostaining.
- Use of scissors and pipettes for tissue preparation.
- Embedding tissue in optimal cutting temperature compound.
- Hematoxylin and eosin staining for tissue architecture assessment.
Main Results
- Well-preserved architecture in various gastric regions.
- Successful immunofluorescent staining of specific cell types.
- Demonstrated expression of key proteins in gastric tissues.
- Confirmed the presence of smooth muscle cells and epithelial cells.
Conclusions
- The Swiss Roll Technique enhances the analysis of gastric tissue.
- It allows for simultaneous processing of all gastric regions.
- This method is practical and scalable for research in health and disease.
