Home › Cell Biology › Visualization of the Interstitial Cells of Cajal (ICC) Network in Mice
Cell Biology JoVE (Open Access) Citable · DOI

Visualization of the Interstitial Cells of Cajal (ICC) Network in Mice

DOI: 10.3791/2802-v
What you'll learn
  • ✓ Dissect and isolate murine gastrointestinal tract tissue
  • ✓ Prepare whole-mount and cryosection samples for immunofluorescence
  • ✓ Perform immunostaining to visualize ICC networks
  • ✓ Interpret fluorescence microscopy images of ICC morphology
Protocol

Biopharma Insights The interstitial cells of Cajal (ICC) are the pacemaker cells of the gastrointestinal (GI) tract. They form complex networks between smooth muscle cells and post-ganglionic neuronal fibers to regulate GI contractility. Here, we present immunofluorescence methods cross-sectional and whole-mount visualization of murine ICC networks.

Difficulty
intermediate
Total time
~3–4 hours per mouse (dissection ~30 min, sample prep and staining ~2–3 hrs, imaging ~30 min)
Model organism
Mouse
Biosafety
BSL-1

Steps

1
Dissect mouse gastrointestinal tract

Remove and isolate the GI tract from euthanized mice using standard dissection techniques. Carefully separate tissue segments for downstream processing.

▶ 01:35
2
Prepare whole-mount tissue and immunostain

Process intact tissue segments using whole-mount immunofluorescence protocols. Incubate with primary and secondary antibodies to label ICC and associated structures.

▶ 02:56
3
Prepare cryosections and immunostain

Generate frozen cross-sections of GI tissue and perform immunofluorescence staining to visualize ICC networks in defined anatomical planes.

▶ 05:10
4
Image and analyze ICC network visualization

Acquire fluorescence microscopy images of both whole-mount and cross-sectional preparations to characterize ICC morphology, distribution, and network connectivity.

▶ 07:44
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