Enhanced Spatial Mapping of Mouse Gastric Muscle Layers Using a Modified Swiss Roll Technique.

Spatial "omics" technologies provide unprecedented opportunities to study the biology and interactions of individual cells of the gastric neuromuscular apparatus in their native tissue environment. However, the successful application of these approaches and the assessment of structural and molecular differences across the fundus, corpus, and antrum require consistent sampling and tissue orientation. Here, we report a modified "Swiss roll" technique tailored to the mouse stomach, which allows all major anatomical regions to be embedded and visualized in a single cryosection. This approach emphasizes flat fixation and careful positioning to generate sections with reproducible orientation and quality. This study also optimized immunofluorescence staining conditions to ensure high-quality visualization of cellular markers. This protocol enables comprehensive, side-by-side comparisons of gastric regions while minimizing tissue loss and experimental variability. By enhancing tissue preservation and spatial orientation, this method offers a practical and scalable tool for researchers studying gastric biology in both physiological and pathological contexts.