The intravital imaging preparation used in this study is similar to previously described methods11 (link),15 (link) with the following differences: imaging is performed with the tissue within the peritoneal cavity, fecal material is not scraped from the mucosal surface and in some experiments atropine (1 mg/kg) was injected subcutaneously to dampen peristaltic movement of the small intestine. At this dose atropine, did not affect the formation of TEDs or GAPs. Model fluorescent antigens, dextran (2–5mg), ovalbumin (2mg), BSA, (2mg) and FluoSpheres (1ml undiluted) (all from Invtirogen, Carlsbad, CA) were injected into the intestinal lumen ~2hrs minutes prior to imaging. Human resection specimens were incubated in 10ug/ml of dextran at room temperature for 1hr prior to imaging.
Protocol detail
Intravital Imaging of Intestinal Antigens
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Antigens
Atropine
Dextran
Fecal
Human
Intestinal
Intestine movement
Mucosal
Ovalbumin
Peristaltic
Peritoneal cavity
Tissue
Corresponding Organization :
Other organizations : Washington University in St. Louis
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Variable analysis
independent variables
- Injection of model fluorescent antigens (dextran, ovalbumin, BSA, FluoSpheres) into the intestinal lumen
- Incubation of human resection specimens in dextran (10ug/ml) for 1 hour prior to imaging
- Formation of TEDs (Transepithelial Dendrites) and GAPs
- Imaging performed with the tissue within the peritoneal cavity
- Fecal material not scraped from the mucosal surface
- Injection of atropine (1 mg/kg) subcutaneously to dampen peristaltic movement of the small intestine
- Positive control: Not specified
- Negative control: Not specified
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