Department of Surgery, Thomas E. Starzl Transplantation Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
Curr Opin Organ Transplant. 2013 Feb;18(1):6-12. doi: 10.1097/MOT.0b013e32835c96c6.
Rejection of transplanted organs is a complex and highly dynamic immune process. Two-photon laser-scanning intravital microscopy (LSIM) allows for real-time, deep tissue, high-resolution imaging in physiological conditions. The recent application of this technology to study organ rejection started to provide a clearer picture of the spatiotemporal immunological dynamics of organ rejection.
To date, LSIM has been applied to transplanted skin, islet, and kidney in mice, as well to constantly moving organs such as transplanted lung and heart. To characterize the dynamics of innate and adaptive immune cell infiltration, time-lapse imaging of various fluorescent-reporter mice was performed. Overall, these studies revealed differences between the anatomical location of infiltrating neutrophils and monocytes in various transplanted organs. In addition, the dynamics of lymphocytic infiltration revealed different transendothelial migration routes in vascularized versus nonvascularized transplanted tissues.
LSIM is a very powerful tool that can be used to carefully dissect the immune cells dynamics in rejection and in tolerance induction in transplantation. Many dynamic biological processes can only be investigated using LSIM. Thus, LSIM promises to greatly enhance our knowledge in transplantation immunobiology and will help tailoring specific therapeutics in relation to the transplanted tissue.
移植器官的排斥是一个复杂且高度动态的免疫过程。双光子激光扫描共聚焦显微镜（LSIM）允许在生理条件下进行实时、深层组织、高分辨率成像。该技术最近被应用于研究器官排斥，开始提供更清晰的器官排斥时空免疫动力学图像。
迄今为止，LSIM 已被应用于小鼠移植的皮肤、胰岛和肾脏，以及不断移动的器官，如移植的肺和心脏。为了描述固有免疫和适应性免疫细胞浸润的动力学，对各种荧光报告小鼠进行了延时成像。总的来说，这些研究揭示了不同移植器官中浸润中性粒细胞和单核细胞的解剖位置的差异。此外，淋巴细胞浸润的动力学揭示了血管化和非血管化移植组织中不同的跨内皮迁移途径。
LSIM 是一种非常强大的工具，可用于仔细剖析排斥反应和移植中诱导耐受过程中的免疫细胞动力学。许多动态生物学过程只能使用 LSIM 进行研究。因此，LSIM 有望极大地提高我们在移植免疫生物学方面的知识，并有助于针对移植组织定制特定的治疗方法。
