Intravital three-photon microscopy allows visualization over the entire depth of mouse lymph nodes.
Intravital three-photon microscopy allows visualization over the entire depth of mouse lymph nodes.
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DOI：
10.1038/s41590-021-01101-1
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发表时间：
2022-03
影响因子：
30.5
通讯作者：
Xu, Chris
中科院分区：
文献类型：
--
作者：
Choe, Kibaek;Hontani, Yusaku;Wang, Tianyu;Hebert, Eric;Ouzounov, Dimitre G.;Lai, Kristine;Singh, Ankur;Beguelin, Wendy;Melnick, Ari M.;Xu, Chris
Intravital confocal microscopy and two-photon microscopy are powerful tools to explore the dynamic behavior of immune cells in mouse lymph nodes (LNs), with penetration depth of ~100 and ~300 μm, respectively. Here, we used intravital three-photon microscopy to visualize the popliteal LN through its entire depth (600–900 μm). We determined the laser average power and pulse energy that caused measurable perturbation in lymphocyte migration. Long-wavelength three-photon imaging within permissible parameters was able to image the entire LN vasculature in vivo and measure CD8+ T cells and CD4+ T cell motility in the T cell zone over the entire depth of the LN. We observed that the motility of naive CD4+ T cells in the T cell zone during lipopolysaccharide-induced inflammation was dependent on depth. As such, intravital three-photon microscopy had the potential to examine immune cell behavior in the deeper regions of the LN in vivo.
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