In vivo confocal endomicroscopy of small intestinal mucosal morphology in dogs.

BACKGROUND Confocal endomicroscopy (CEM) is an endoscopic technology that permits in vivo cellular and subcellular imaging of the gastrointestinal mucosa. OBJECTIVE To determine the feasibility of CEM to evaluate small intestinal mucosal topologic morphology in dogs and to characterize the appearance in healthy dogs. ANIMALS Fourteen clinically healthy research colony dogs. METHODS Experimental study. Dogs were anesthetized for standard endoscopic evaluation of the small intestine followed by CEM. Two fluorophores were used to provide contrast: fluorescein (10% solution, 15 mg/kg IV) before administration of topical acriflavine (0.05% solution) via an endoscopy spray catheter. A minimum of 5 sites within the small intestine were assessed and at each location, sequential adjustment of imaging depth allowed collection of a three-dimensional volume equivalent to an 'optical biopsy'. CEM-guided pinch biopsies were obtained for histologic examination. RESULTS CEM provided high-quality in vivo cellular and subcellular images. Intravenous administration of fluorescein provided sufficient contrast to allow assessment of the vasculature, cellular cytoplasmic features and goblet cell numbers, and distribution. Topical application of acriflavine preferentially stained cellular nucleic acids, allowing evaluation of nuclear morphology. Quality of captured images was occasionally affected by motion artifact, but improved with operator experience. CONCLUSION AND CLINICAL IMPORTANCE CEM provides in vivo images that allow for cellular and subcellular assessment of intestinal mucosal morphology during endoscopy. This has implications for aiding in vivo diagnosis of gastrointestinal disease.

[1]  A. Watson,et al.  Epithelial barrier function in vivo is sustained despite gaps in epithelial layers. , 2005, Gastroenterology.

[2]  Andrew V Biankin,et al.  In vivo confocal endomicroscopy in the diagnosis and evaluation of celiac disease. , 2008, Gastroenterology.

[3]  G. D. De Palma,et al.  Confocal laser endomicroscopy in the "in vivo" histological diagnosis of the gastrointestinal tract. , 2009, World journal of gastroenterology.

[4]  Michael Vieth,et al.  Confocal Laser Endomicroscopy , 2012, Gastroenterology research and practice.

[5]  Simon K Lo,et al.  The learning curve of in vivo probe-based confocal laser endomicroscopy for prediction of colorectal neoplasia. , 2009, Gastrointestinal endoscopy.

[6]  A. Jergens,et al.  Endoscopic, biopsy, and histopathologic guidelines for the evaluation of gastrointestinal inflammation in companion animals. , 2010, Journal of veterinary internal medicine.

[7]  M. Canto,et al.  Confocal endomicroscopy , 2008, Current opinion in gastroenterology.

[8]  Yan-Qing Li,et al.  Classification of gastric pit patterns by confocal endomicroscopy. , 2008, Gastrointestinal endoscopy.

[9]  N. Nguyen,et al.  Current application of confocal endomicroscopy in gastrointestinal disorders , 2008, Journal of gastroenterology and hepatology.

[10]  S. Brookes,et al.  Expression of Notch1 and Jagged2 in the Enteric Nervous System , 2003, Journal of Histochemistry and Cytochemistry.

[11]  T. Whittem,et al.  The exogenous fluorophore, fluorescein, enables in vivo assessment of the gastrointestinal mucosa via confocal endomicroscopy: optimization of intravenous dosing in the dog model. , 2013, Journal of veterinary pharmacology and therapeutics.

[12]  R. Kiesslich,et al.  In-vivo confocal real-time mini-microscopy in animal models of human inflammatory and neoplastic diseases. , 2007, Endoscopy.

[13]  H. Neuhaus,et al.  Endomicroscopy of Gastritis and Gastric Cancer , 2008 .

[14]  R. Kiesslich,et al.  Simultaneous confocal laser endomicroscopy and chromoendoscopy with topical cresyl violet. , 2009, Gastrointestinal endoscopy.

[15]  Giovanni Domenico De Palma,et al.  Confocal laser endomicroscopy in the "in vivo" histological diagnosis of the gastrointestinal tract. , 2009 .

[16]  M. Goetz,et al.  Examination technique of confocal laser endomicroscopy , 2008 .

[17]  R. Kiesslich,et al.  Confocal laser endomicroscopy for gastrointestinal diseases. , 2008, Gastrointestinal endoscopy clinics of North America.

[18]  A. Bardai,et al.  Fluorogenic Substrate [Ala-Pro]2-Cresyl Violet But Not Ala-Pro-Rhodamine 110 Is Cleaved Specifically by DPPIV Activity: A Study in Living Jurkat Cells and CD26/DPPIV-transfected Jurkat Cells , 2003, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[19]  Barry Campbell,et al.  Confocal laser endomicroscopy is a new imaging modality for recognition of intramucosal bacteria in inflammatory bowel disease in vivo , 2010, Gut.

[20]  G. Thiessen,et al.  The acridine dyes: Their purification, physicochemical, and cytochemical properties , 1976, Histochemistry.

[21]  Taking optical biopsies with confocal endomicroscopy , 2009, Journal of gastroenterology and hepatology.

[22]  A. Polglase,et al.  Confocal laser endoscopy for diagnosing intraepithelial neoplasias and colorectal cancer in vivo. , 2004, Gastroenterology.

[23]  P. Delaney,et al.  In vivo mapping of the vascular changes in skin burns of anaesthetised mice by fibre optic confocal imaging (FOCI). , 2000, Journal of dermatological science.

[24]  Steffen Jung,et al.  Quantitative analysis of intravenously administered contrast media reveals changes in vascular barrier functions in a murine colitis model , 2011, Magnetic resonance in medicine.

[25]  Confocal endomicroscopy for the diagnosis of gastric cancer in vivo , 2007 .

[26]  O. Maeda,et al.  Confocal endomicroscopy in patients with ulcerative colitis , 2008, Journal of gastroenterology and hepatology.

[27]  G. Moore,et al.  Effect of tissue processing on assessment of endoscopic intestinal biopsies in dogs and cats. , 2010, Journal of veterinary internal medicine.

[28]  C. Crosta,et al.  Celiac disease: in vivo diagnosis by confocal endomicroscopy. , 2007, Gastrointestinal endoscopy.

[29]  E. Montgomery,et al.  Confocal Laser Endomicroscopy Image Interpretation: Interobserver Agreement Among Gastroenterologists and Pathologists , 2007 .