Visualization of mucosal homeostasis via single- and multiphoton intravital fluorescence microscopy

Journal of Leukocyte Biology

Cassie Xu²  Yuelei Shen³  Dan R. Littman⁵  Michael L. Dustin and⁴  Peter VelÃzquez,–¹ 

[1] Department of Microbiology and Immunology, Indiana University School of Medicine, South Bend, Indiana, USA; The Kimmel Center for Biology and Medicine of the Skirball Institute, New York, New York, USA; and Department of Biological Sciences and –Eck Institute for Global Health, University of Notre Dame, Notre Dame, Indiana, USADepartment of Microbiology and Immunology, Indiana University School of Medicine, South Bend, Indiana, USA; Howard Hughes Medical Institute, New York University School of Medicine, New York, New York, USA;The Kimmel Center for Biology and Medicine of the Skirball Institute, New York, New York, USA; Howard Hughes Medical Institute, New York University School of Medicine, New York, New York, USA; The Kimmel Center for Biology and Medicine of the Skirball Institute, New York, New York, USA;

关键词: T cells;    dendritic cells;    intestine;   

DOI  :  10.1189/jlb.0711344

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

FIVM has provided many insights into the regulation of immunity. We report the validation of an approach for visualizing murine small bowel via single- and multiphoton FIVM. Tissue damage is limited to ∼200 μm, immediately adjacent to the incision, as confirmed by intravital PI staining. Treatment with 10 KDa dextran-FITC and 70 KDa dextran-TR confirms that perfusion is intact. Selective filtration of 10 KDa but not 70 KDa dextran from the blood indicated that kidney function is also intact. Interestingly, lamina propria vasculature is semipermeable to 10 KDa dextran. Next, reporter mice expressing egfp from the CX3CR1 locus, egfp from the FoxP3 locus, or RFP from the IL-17F locus were used to track DC subsets, FoxP3+ Tregs, or Th17f cells, respectively. Resident cx3cr1+/egfp cells were sessile but actively probed the surrounding microenvironment. Both T cell populations patrol the lamina propria, but the Th17f cells migrate more rapidly than Tregs. Together, these data demonstrate intact vascular perfusion, while intravitally visualizing the mucosal surface of the small bowel. Lastly, the cx3cr1+ DCs and T cells display activity similar to that found in steady-state, secondary lymphoid organs.

【 授权许可】

Unknown