F (Fig 1). Figure 1 Preparation of the rats for intraoperative endoscopy, and the experimental setup. Laparotomy was performed in WAG/Rij rats which were under inhalation narcosis. The opened cecum was positioned with the lumen upwards on a specially designed small table … Following the procedure all animals were sacrificed immediately using T61 (Intervet, Germany). Endoscopes and confocal laser microscopy For endoscopy, a Lucera CF-H260AZL/I® Inhibitors,research,lifescience,medical colonoscope attached to the EVIS Lucera Spectrum® video system (Olympus, Tokyo, Japan) was employed,
providing 1080i high-definition television (HDTV) imaging and two additional light observation modes, that is, narrow-band imaging (NBI) and autofluorescence imaging (AFI). For confocal laser microscopy (CLM) a Leica animal Inhibitors,research,lifescience,medical Z-probe attached to a laser scanning unit (Cellvizio®-Lab system; Mauna Kea Technologies, Paris, France) was used. Leica FCM 1000 IC software
was employed for image documentation. Immediately before the CLM examination, 0.1 ml Inhibitors,research,lifescience,medical of a 5% fluorescein solution (Alcon Pharma GmbH, Freiburg, Germany) was injected into the rat’s tail vein. Results Preparation of the animal model Sufficient inhalation anesthesia of an individual animal, opening of the abdominal cavity, longitudinal exposure of the cecum and cleansing of stool required a mean time of 11 minutes (SD ±4 min). The integrity of the blood flow was determined by i.v. administration of Evans blue dye (Sigma-Aldrich, Munich, Germany) through the tail vein of a tumor-free WAG/Rij rat, revealing an apparently intact blood www.selleckchem.com/products/BI6727-Volasertib.html supply for the maximum tested time of 90 minutes (Fig 2). The adequacy of the blood supply was also proven by CLM imaging Inhibitors,research,lifescience,medical after i.v. administration of fluorescein in all animals. Moreover, no notable bleeding occurred in any of the animals. Figure 2 Integrity of the intestinal blood supply. Preservation of the intestinal blood supply Inhibitors,research,lifescience,medical was tested
by tail vein injection of Evans blue dye which resulted in a blue coloration. The arrows mark the lumen of the exposed bowel, and the outside of the bowel … The opened cecum, placed on a small table located CYTH4 above the rat’s chest, offered an area of approximately 4 × 2 cm of exposed mucosa for further endoscopic evaluation. All animals that had undergone prior injection of the R1H cell line displayed tumor nodes of 3–8 mm diameter at the injection sites. Assessment of NBI, AFI, and CLM in rat colon tumors To test the applicability of a clinical-scale endoscope to inspect the rat cecum, a colonoscope was placed close to the mucosa. Conventional white-light endoscopy showed that about two thirds of the cecum could be observed while keeping a fixed position, with slight movements of the endoscope tip allowing gradual inspection of the entire surface.