Artikel
An unbiased approach to analysis of experimental colitis
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Veröffentlicht: | 24. April 2015 |
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Gliederung
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Introduction: Animal models of intestinal inflammation are extensively employed in research of inflammatory bowel diseases. However histological evaluation can be limited by a degree of subjectivity leading to variable results in the literature. Here we describe an unbiased widely applicable histological protocol for reproducible quantification of intestinal mucosal damage in murine models of intestinal inflammation.
Material and methods: High resolution digital scanning of the tissue and computer assisted analysis was used to evaluate the percentage of mucosal inflammation, injury and ulceration at a ratio of the whole colon length allowing for precise and unbiased data acquisition. For image analysis of scanned histological slides, the freely accessible program Aperio Imagescope was used. The results have been validated with other disease markers such as measurement of myeloperoxidase activity, fecal lipocalin concentration (ELISA) or the murine endoscopic index for colitis severity (MEICS) using mouse colonoscopy.
Results: The protocol has been established for DSS and T-cell transfer models of intestinal inflammation/injury. We show that the percentage of inflammation/ulceration in ratio to the whole colon length faithfully reflects the percentage of weight loss during the course of the disease and therefore is a valuable surrogate marker for disease activity. Furthermore the percentage of ulceration highly correlates with the evaluated other markers for inflammation such as the murine endoscopic index for colitis severity.
Conclusion: The presented protocol gives comprehensive information about sample preparation, image analysis and data presentation. Using the proposed method will result in high objectivity and reproducibility of the results.
Figure 1 [Fig. 1]