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Updated: May 11, 2026
Imaging CD4 T Cell Interstitial Migration in the Inflamed Dermis
Published on: March 25, 2016
Live cell imaging of chemotactic dendritic cell migration in explanted mouse ear preparations
1Institute of Science and Technology Austria, Klosterneuburg, Austria.
Insights
Researchers developed a new ex vivo assay using mouse ears to visualize chemokine-guided leukocyte migration in real time. This method offers a more physiological approach to study immune cell trafficking in the dermal interstitium.
Area of Science:
- Immunology
- Cell Biology
- Biophysics
Background:
- Leukocyte migration is vital for immune responses and maintaining tolerance.
- Chemokines are key signaling molecules guiding leukocyte trafficking in vivo.
- Existing in vitro models lack the physiological relevance for studying cell migration in complex tissues.
Purpose of the Study:
- To develop and validate a novel ex vivo assay for quantifying chemokine-guided leukocyte migration.
- To visualize and analyze dendritic cell (DC) motility within the dermal interstitium in real time.
- To provide a more physiologically relevant model compared to traditional 2D and 3D in vitro assays.
Main Methods:
- Preparation of explanted mouse ear sheets.
- Utilizing multidimensional confocal imaging for real-time monitoring.
- Tracking fluorescently labeled dendritic cells (DCs) in the dermal interstitium and lymphatic vessels.
Main Results:
- Successful visualization of chemokine-dependent DC migration in a physiologically relevant ex vivo setting.
- Quantification of DC motility within the dermal interstitium.
- Observation of DC entry into afferent lymphatic vessels.
Conclusions:
- The ex vivo mouse ear crawl-in assay is a powerful tool for studying leukocyte migration dynamics.
- This assay offers superior physiological relevance for investigating immune cell trafficking.
- The developed methods enable real-time analysis of DC behavior in response to chemokines.
Abstract:
Leukocyte migration through the interstitial space is crucial for the maintenance of tolerance and immunity. The main cues for leukocyte trafficking are chemokines thought to directionally guide these cells towards their targets. However, model systems that facilitate quantification of chemokine-guided leukocyte migration in vivo are uncommon. Here we describe an ex vivo crawl-in assay using explanted mouse ears that allows the visualization of chemokine-dependent dendritic cell (DC) motility in the dermal interstitium in real time. We present methods for the preparation of mouse ear sheets and their use in multidimensional confocal imaging experiments to monitor and analyze the directional migration of fluorescently labelled DCs through the dermis and into afferent lymphatic vessels. The assay provides a more physiological approach to study leukocyte migration than in vitro three-dimensional (3D) or 2-dimensional (2D) migration assays such as collagen gels and transwell assays.
