Abstract
Intravital microscopy is an imaging technique aimed at the visualization of the dynamics of biological processes in live animals. In the last decade, the development of nonlinear optical microscopy has enormously increased the use of this technique, thus addressing key biological questions in different fields such as immunology, neurobiology and tumor biology. In addition, new upcoming strategies to minimize motion artifacts due to animal respiration and heartbeat have enabled the visualization in real time of biological processes at cellular and subcellular resolution. Recently, intravital microscopy has been applied to analyze different aspect of mucosal immunity in the gut. However, the majority of these studies have been performed on the small intestine. Although crucial aspects of the biology of this organ have been unveiled, the majority of intestinal pathologies in humans occur in the large intestine.
Here, we describe a method to surgically expose and stabilize the large intestine in live mice and to perform short-term (up to 2 h) intravital microscopy.
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Change history
23 July 2021
Chapter 15 “Method for Acute Intravital Imaging of the Large Intestine in Live Mice” was previously published with incorrect ESM videos and figure captions. This has now been rectified in the revised version of this book.
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Acknowledgments
This research was supported by the Intramural Research Program of the National Institutes of Health, National Cancer Institute, Center for Cancer Research (RW). This work has been supported by a UICC International Cancer Research Technology Transfer Fellowship (UICC ICRETT ICR/2015/40447 fellowship to ME).
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1 Electronic Supplementary Materials
Blood flow in the large intestine mucosal layer. Green: FITC-conjugated dextran. Scale bar: 20 μm. The original pixel size has been computationally modified for a better video quality. (AVI 1223 kb)
Z-stack acquisition of the large intestine of B5/EGFP mice, from the muscular layer to the mucosal layer. Scale bar: 100 μm. (AVI 4461 kb)
Blood flow in the large intestine muscular layer. Red: TRITC-conjugated dextran, Green: CX3CR1gfp- positive tissue macrophages. Scale bar: 50 μm. The original pixel size has been computationally modified for a better video quality. (AVI 372 kb)
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Erreni, M., Doni, A., Weigert, R. (2021). Method for Acute Intravital Imaging of the Large Intestine in Live Mice. In: Brzostowski, J., Sohn, H. (eds) Confocal Microscopy. Methods in Molecular Biology, vol 2304. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1402-0_15
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DOI: https://doi.org/10.1007/978-1-0716-1402-0_15
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