Abstract
The murine intestinal tract represents a difficult organ system to study due to its long convoluted tubular structure, narrow diameter, and delicate mucosa which undergoes rapid changes after sampling prior to fixation. These features do not make for easy histological analysis as rapid fixation in situ, or after simple removal without careful dissection, results in poor postfixation tissue handling and limited options for high quality histological sections. Collecting meaningful quantitative data by analysis of this tissue is further complicated by the anatomical changes in structure along its length. This article describes two methods of intestinal sampling at necropsy that allow systematic histological analysis of the entire intestinal tract, either through examination of cross sections (circumferences) by the gut bundling technique or longitudinal sections by the adapted Swiss roll technique, together with basic methods for data collection. © 2016 by John Wiley & Sons, Inc.
Highlights
This unit details the techniques that may be employed for histological assessment of the murine intestinal tract
Anticipated Results: The anticipated results for the outlined methods are multiple high quality, well preserved intestinal sections which may be stained by hematoxylin and eosin (H&E) or with immunohistochemical stains
This should allow both qualitative and quantitative data to be obtained from these tissues in the context of mouse models of intestinal diseases
Summary
This unit details the techniques that may be employed for histological assessment of the murine intestinal tract. If luminal content is required (for example for microbiome studies), this can be incorporated into the gut bundling technique or the Swiss roll method, by sampling of content from specific parts of the gastrointestinal tract prior to fixation This can be achieved by making a small incision in the intestinal wall with scissors. To assess features such as villus tip apoptosis (Williams et al, 2013) extra care must be taken to rapidly fix the small intestine, as the villus tip is the most susceptible to artefactual changes For studies such as this, the gut bundling method is preferred over the Swiss roll technique due to the rapid immersion in formalin with less tissue handling, less compression and distortion of villi, and preservation of apoptotic cells in the lumen. If the stomach is to be taken for histological analysis and prepared by instillation with fixative, the pylorus can be located and ligated at this point with 4 metric sofsilkTM suture material (see Duckworth et al, 2015 for detailed description of protocol)
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