Protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons
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Updated: Jun 12, 2025
In Situ Ca2+ Imaging of the Enteric Nervous System
Published on: January 29, 2015
Egan L Choi1, Yuebo Zhang1, Fei Gao1
1Enteric Neuroscience Program and Department of Physiology and Biomedical Engineering, Mayo Clinic College of Medicine and Science, Rochester, MN 55905, USA; Gastroenterology Research Unit, Division of Gastroenterology and Hepatology, Department of Medicine, Mayo Clinic College of Medicine and Science, Rochester, MN 55905, USA.
Abstract:
Gastrointestinal motor functions are orchestrated by enteric neurons that display a high degree of anatomical and functional variability. Here, we present a protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons. We describe steps for human gastric tunica muscularis preparation, immunofluorescent labeling of enteric neurons, and quantification of marker colocalization using Nikon NIS-Elements. This protocol addresses the challenges presented by the high density of neurons within enteric ganglia, overlapping fluorescence signals, background fluorescence, and potential user bias in quantification. For complete details on the use and execution of this protocol, please refer to Gao et al.1.
