Protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons

Egan L. Choi; Yuebo Zhang; Fei Gao; Nick M. Huynh; Alejandro Stark Quiroz; Adil E. Bharucha; Yujiro Hayashi; Tamas Ordog

doi:10.1016/j.xpro.2025.103828

Protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons

Egan L. Choi, Yuebo Zhang, Fei Gao, Nick M. Huynh, Alejandro Stark Quiroz, Adil E. Bharucha, Yujiro Hayashi, Tamas Ordog

Research output: Contribution to journal › Article › peer-review

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

Original language	English (US)
Article number	103828
Journal	STAR Protocols
Volume	6
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2025.103828
State	Published - Jun 20 2025

Keywords

Microscopy
Molecular Biology
Neuroscience

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

Access to Document

10.1016/j.xpro.2025.103828

Cite this

@article{707896631a9a4e9ca7d59174cde50daf,

title = "Protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons",

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",

keywords = "Microscopy, Molecular Biology, Neuroscience",

author = "Choi, \{Egan L.\} and Yuebo Zhang and Fei Gao and Huynh, \{Nick M.\} and Quiroz, \{Alejandro Stark\} and Bharucha, \{Adil E.\} and Yujiro Hayashi and Tamas Ordog",

note = "Publisher Copyright: {\textcopyright} 2025 The Author(s)",

year = "2025",

month = jun,

day = "20",

doi = "10.1016/j.xpro.2025.103828",

language = "English (US)",

volume = "6",

journal = "STAR Protocols",

issn = "2666-1667",

publisher = "Cell Press",

number = "2",

}

TY - JOUR

T1 - Protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons

AU - Choi, Egan L.

AU - Zhang, Yuebo

AU - Gao, Fei

AU - Huynh, Nick M.

AU - Quiroz, Alejandro Stark

AU - Bharucha, Adil E.

AU - Hayashi, Yujiro

AU - Ordog, Tamas

PY - 2025/6/20

Y1 - 2025/6/20

N2 - 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

AB - 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

KW - Microscopy

KW - Molecular Biology

KW - Neuroscience

UR - https://www.scopus.com/pages/publications/105005170578

UR - https://www.scopus.com/inward/citedby.url?scp=105005170578&partnerID=8YFLogxK

U2 - 10.1016/j.xpro.2025.103828

DO - 10.1016/j.xpro.2025.103828

M3 - Article

AN - SCOPUS:105005170578

SN - 2666-1667

VL - 6

JO - STAR Protocols

JF - STAR Protocols

IS - 2

M1 - 103828

ER -

Protocol for AI-supported immunofluorescence colocalization analysis in human enteric neurons

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this