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Updated: Jun 12, 2025
In Situ Ca2+ Imaging of the Enteric Nervous System
Published on: January 29, 2015
Protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons.
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.
This study introduces an AI-powered protocol for analyzing human enteric neurons using immunofluorescence. It overcomes challenges in quantifying neuronal markers in dense tissues, aiding gastrointestinal research.
Area of Science:
- Neuroscience
- Gastroenterology
- Biotechnology
Background:
- Gastrointestinal motor functions rely on complex enteric nervous systems.
- Enteric neurons exhibit significant anatomical and functional diversity.
- Accurate analysis of enteric neurons is crucial for understanding gut motility.
Purpose of the Study:
- To present a protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons.
- To provide a standardized method for quantifying neuronal marker expression.
- To address challenges in analyzing dense enteric ganglia.
Main Methods:
- Human gastric tunica muscularis preparation.
- Immunofluorescent labeling of enteric neurons.
- AI-assisted quantification of marker colocalization using Nikon NIS-Elements software.
Main Results:
- The protocol effectively quantifies marker colocalization in human enteric neurons.
- It mitigates issues like high neuronal density, signal overlap, and background noise.
- The method reduces potential user bias in quantitative analysis.
Conclusions:
- This AI-driven protocol offers a robust approach for analyzing human enteric neurons.
- It facilitates a deeper understanding of the anatomical and functional variability of these neurons.
- The protocol is a valuable tool for research in gastrointestinal neurobiology and motility disorders.
