Abstract
T-cell motility is essential for the T cells’ ability to scan antigens within lymph nodes and initiate contact with antigen-presenting cells. While T-cell migration has been extensively studied using in vitro migration assays, accumulating evidence indicates that the T-cell migration within lymph nodes is modulated by the surrounding cells and extracellular matrix, which form the confined architecture of the lymph nodes. Therefore, to understand the mechanisms of T-cell motility in vivo, their cell migration must be analyzed under physiological conditions. To this end, two-photon microscopy is extremely useful; this technique enables the tracking of fluorescently labeled cells in vivo and ex vivo, with high spatial and temporal resolutions. Here we describe the experimental procedures for applying two-photon microscopy to the in vivo and ex vivo imaging of T-cell migration in mouse lymph nodes. These approaches provide physiological insight into the mechanisms of T-cell behavior at a single-cell level in the three-dimensional lymph node environment.
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Acknowledgements
We thank Mr. Keita Aoi and Prof. Masaru Ishii (Osaka University) for their helpful advice in establishing procedures for the two-photon imaging of LNs.
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Takeda, A., Miyasaka, M., Umemoto, E. (2018). Two-Photon Imaging of T-Cell Motility in Lymph Nodes: In Vivo and Ex Vivo Approaches. In: Ishii, M. (eds) Intravital Imaging of Dynamic Bone and Immune Systems . Methods in Molecular Biology, vol 1763. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7762-8_5
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DOI: https://doi.org/10.1007/978-1-4939-7762-8_5
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