Imaging inflammation in mouse colon using a rapid stage-scanning confocal fluorescence microscope.

Large area confocal microscopy may provide fast, high-resolution image acquisition for evaluation of tissue in pre-clinical studies with reduced tissue processing in comparison to histology. We present a rapid beam and stage-scanning confocal fluorescence microscope to image cellular and tissue features along the length of the entire excised mouse colon. The beam is scanned at 8,333 lines/sec by a polygon scanning mirror while the specimen is scanned in the orthogonal axis by a motorized translation stage with a maximum speed of 7 mm/sec. A single 1 × 60 mm(2) field of view image spanning the length of the mouse colon is acquired in 10 s. Z-projection images generated from axial image stacks allow high resolution imaging of the surface of non-flat specimens. In contrast to the uniform size, shape, and distribution of colon crypts in confocal images of normal colon, confocal images of chronic bowel inflammation exhibit heterogeneous tissue structure with localized severe crypt distortion.

[1]  Milind Rajadhyaksha,et al.  Rapid confocal imaging of large areas of excised tissue with strip mosaicing. , 2011, Journal of biomedical optics.

[2]  T. Wilson,et al.  Optical sectioning in fluorescence microscopy , 2011, Journal of microscopy.

[3]  Euiheon Chung,et al.  In vivo wide-area cellular imaging by side-view endomicroscopy , 2010, Nature Methods.

[4]  Yogesh G. Patel,et al.  Confocal mosaicing microscopy in skin excisions: a demonstration of rapid surgical pathology , 2009, Journal of microscopy.

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

[6]  P. Maisonneuve,et al.  Screening and surveillance for the early detection of colorectal cancer and adenomatous polyps. , 2008, Gastroenterology.

[7]  J. Lupton,et al.  Reduced colitis-associated colon cancer in Fat-1 (n-3 fatty acid desaturase) transgenic mice. , 2008, Cancer research.

[8]  J. Lupton,et al.  Mechanisms by which docosahexaenoic acid and related fatty acids reduce colon cancer risk and inflammatory disorders of the intestine. , 2008, Chemistry and physics of lipids.

[9]  B. Levin,et al.  Screening and surveillance for the early detection of colorectal cancer and adenomatous polyps, 2008: a joint guideline from the American Cancer Society, the US Multi-Society Task Force on Colorectal Cancer, and the American College of Radiology. , 2008, Gastroenterology.

[10]  E. Loftus The burden of inflammatory bowel disease in the United States: a moving target? , 2007, Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association.

[11]  Ann M Gillenwater,et al.  Dual‐mode reflectance and fluorescence near‐video‐rate confocal microscope for architectural, morphological and molecular imaging of tissue , 2007, Journal of microscopy.

[12]  Yogesh G. Patel,et al.  Confocal reflectance mosaicing of basal cell carcinomas in Mohs surgical skin excisions. , 2007, Journal of biomedical optics.

[13]  P. Mannon,et al.  The fundamental basis of inflammatory bowel disease. , 2007, The Journal of clinical investigation.

[14]  Robert S Chapkin,et al.  Colon cancer, fatty acids and anti-inflammatory compounds , 2007, Current opinion in gastroenterology.

[15]  R. Kiesslich,et al.  Confocal laser endomicroscopy: technical status and current indications , 2006, Endoscopy.

[16]  Alicja Wolk,et al.  Meat consumption and risk of colorectal cancer: A meta‐analysis of prospective studies , 2006, International journal of cancer.

[17]  Christopher H Contag,et al.  Dual-axes confocal reflectance microscope for distinguishing colonic neoplasia. , 2006, Journal of biomedical optics.

[18]  R. Sinha,et al.  Meat consumption and risk of colorectal cancer. , 2005, JAMA.

[19]  S. Itzkowitz,et al.  Inflammation and cancer IV. Colorectal cancer in inflammatory bowel disease: the role of inflammation. , 2004, American journal of physiology. Gastrointestinal and liver physiology.

[20]  Takuji Tanaka,et al.  A novel inflammation‐related mouse colon carcinogenesis model induced by azoxymethane and dextran sodium sulfate , 2003, Cancer science.

[21]  W B Amos,et al.  How the Confocal Laser Scanning Microscope entered Biological Research , 2003, Biology of the cell.

[22]  H. Cooper,et al.  Clinicopathologic study of dextran sulfate sodium experimental murine colitis. , 1993, Laboratory investigation; a journal of technical methods and pathology.

[23]  A. Medline,et al.  Dose response and proliferative characteristics of aberrant crypt foci: putative preneoplastic lesions in rat colon. , 1991, Carcinogenesis.

[24]  D. Podolsky Inflammatory bowel disease (Second of two parts) , 1991 .

[25]  D. Levine,et al.  Normal histology of the colon. , 1989, The American journal of surgical pathology.

[26]  T. Wilson,et al.  Optical sectioning in confocal fluorescent microscopes , 1989 .

[27]  M. Fordham,et al.  An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy , 1987, The Journal of cell biology.

[28]  Carol M. Park,et al.  A simple, practical ‘swiss roll’ method of preparing tissues for paraffin or methacrylate embedding , 1987, Journal of microscopy.

[29]  G. J. Brakenhoff,et al.  Confocal scanning light microscopy with high aperture immersion lenses , 1979 .

[30]  P. Davidovits,et al.  Scanning Laser Microscope , 1969, Nature.

[31]  E. Szigethy,et al.  Inflammatory bowel disease. , 2011, Pediatric clinics of North America.

[32]  Caroline Boudoux,et al.  Comprehensive imaging of gastroesophageal biopsy samples by spectrally encoded confocal microscopy. , 2010, Gastrointestinal endoscopy.

[33]  J. Pawley,et al.  Handbook of Biological Confocal Microscopy , 1990, Springer US.

[34]  M. Minsky Memoir on inventing the confocal scanning microscope , 1988 .