Generation of Mouse Proximal Colon Loop Model: A Surgical Procedure to Generate Proximal Colon Loop and Measure Intestinal Permeability by Detection of Fluorescent Markers in Serum Following Intraluminal Injection
Published: April 30, 2023
Abstract
Source: Boerner, K. et al. Functional Assessment of Intestinal Permeability and Neutrophil Transepithelial Migration in Mice using a Standardized Intestinal Loop Model. J. Vis. Exp. (2021).
In this video, we describe a surgical procedure to generate mouse proximal colon loop or pcLoop segment followed by intraluminal injection of fluorescent markers into the segment to study the intestinal epithelial permeability.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Generation of the proximal colon loop (pcLoop) Skin preparation: Scrub fur of the abdominal midline with alcohol swabs or gauze sponge soaked with 70% Ethanol. Do not wet a wide area of fur with alcohol to prevent hypothermia. Using scissors, perform a midline laparotomy. Make a vertical incision in the middle…
Representative Results
Figure 1: The proximal colon loop model. (A) Schematic overview of the pcLoop model. Median laparotomy is performed on mice under anesthesia placed on a temperature-controlled surgery board. (B) Exteriorization of the caecum (*), proximal colon, mesocolon and ileum. Two adequate sites for ligation are identified (1,2). (C) The first ligature (1) …
Offenlegungen
The authors have nothing to disclose.
Materials
| Equipment and Material | |||
| BD Alcohol Swabs | BD | 326895 | |
| BD 1ml Tuberculin Syringe Without Needle | BD | 309659 | |
| BD PrecisionGlide Needle, 25G X 5/8" | BD | 305122 | |
| BD PrecisionGlide Needle, 30G X 1/2" | BD | 305106 | |
| Cotton Tip Applicator (cotton swab), 6", sterile | FisherScientific | 25806 2WC | |
| Dynarex Cotton Filled Gauze Sponges, Non-Sterile, 2" x 2" | Medex | 3249-1 | |
| EZ-7000 anesthesia vaporizer (Classic System, including heating units) | E-Z Systems | EZ-7000 | |
| Kimwipes, small (tissue wipe) | FisherScientific | 06-666 | |
| Moria Fine Scissors | FST | 14370-22 | |
| Puralube Vet Ointment, Sterile Ocular Lubricant | Dechra | 12920060 | |
| Ring Forceps (blunt tissue forceps) | FST | 11103-09 | |
| Roboz Surgical 4-0 Silk Black Braided, 100 YD | FisherScientific | NC9452680 | |
| Semken Forceps (anatomical forceps) |
FST | 1108-13 | |
| Sofsilk Nonabsorbable Coated Black Suture Braided Silk Size 3-0, 18", Needle 19mm length 3/8 circle reverse cutting | HenrySchein | SS694 | |
| Student Fine Forceps, Angled | FST | 91110-10 | |
| 10ml Syringe PP/PE without needle | Millipore Sigma | Z248029 | |
| Reagents | |||
| Isoflurane | Halocarbon | 12164-002-25 | |
| Fluorescein Isothiocyanate-Dextran, average molecular weight 4.000 | Sigma | 60842-46-8 |
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Generation of Mouse Proximal Colon Loop Model: A Surgical Procedure to Generate Proximal Colon Loop and Measure Intestinal Permeability by Detection of Fluorescent Markers in Serum Following Intraluminal Injection. J. Vis. Exp. (Pending Publication), e20813, doi: (2023).