Protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons
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.<sup>1</sup>.
Authors
- Yuebo Zhang
- Fei Gao (ORCID: https://orcid.org/0000-0003-2291-8712)
- Tamás Ördög (ORCID: https://orcid.org/0000-0002-3940-7284)
- Adil E. Bharucha (ORCID: https://orcid.org/0000-0002-7644-0390)
- Yujiro Hayashi (ORCID: https://orcid.org/0000-0002-4555-2285)
- Egan L. Choi (ORCID: https://orcid.org/0000-0002-1258-0896)
- Nick M. Huynh
- Alejandro Stark Quiroz
Institutions
- Mayo Clinic (US)
- Mayo Clinic in Arizona (US)
- Mayo Clinic in Florida (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-05-16
- DOI
- https://doi.org/10.1016/j.xpro.2025.103828
- Citations
- 1
- Primary Topic
- Congenital gastrointestinal and neural anomalies
- Type
- article
- Field-Weighted Citation Impact
- 2.01
Funders
- Mayo Clinic
- National Institutes of Health
- National Institute of Diabetes and Digestive and Kidney Diseases