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Related Experiment Video

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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.

STAR Protocols
|May 18, 2025
PubMed
Summary

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.

Keywords:
MicroscopyMolecular BiologyNeuroscience

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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.