Multiphoton Intravital Microscopy of the Transplanted Mouse Kidney
Multiphoton Intravital Microscopy of the Transplanted Mouse Kidney
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DOI：
10.1111/j.1600-6143.2011.03671.x
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发表时间：
2011-10-01
影响因子：
8.8
通讯作者：
Lakkis, F. G.
中科院分区：
文献类型：
--
作者：
Camirand, G.;Li, Q.;Lakkis, F. G.
肾移植后的移植结果继续受到缺血-再灌注损伤和排斥反应的不利影响。移植肾的高分辨率、实时成像可以为这些动态过程提供有价值的见解，但这种方法尚未建立。在这里，我们描述了一种使用多光子荧光显微镜对移植的小鼠肾脏进行活体成像的技术。这项技术能够实时、高分辨率地成像和量化移植后的肾滤过、细胞死亡、白细胞黏附和毛细血管血流。利用这项技术，我们发现与移植程序相关的短暂移植物缺血导致移植后3h内肾滤过迅速下降，并伴随着微血管渗漏和肾小管上皮细胞死亡的显著增加。在同一时间段内，未观察到白细胞黏附和毛细血管血流量的明显变化。这份报告建立了多光子荧光显微镜作为一种灵敏的工具，用于同时研究移植后小鼠肾脏发生的功能和结构扰动，并研究白细胞向移植物的迁移。
Graft outcomes after kidney transplantation continue to be adversely affected by ischemia-reperfusion injury and rejection. High-resolution, real-time imaging of the transplanted kidney could shed valuable insights into these dynamic processes, but such methodology has not been established. Here we describe a technique for intravital imaging of the transplanted mouse kidney using multiphoton fluorescence microscopy. The technique enabled real-time, high-resolution imaging and quantitation of renal filtration, cell death, leukocyte adhesion and capillary blood flow after transplantation. Using this technique, we found that brief graft ischemia associated with the transplantation procedure led to a rapid decline in renal filtration accompanied by a significant increase in microvascular leakage and renal tubular epithelial cell death within the first 3 h after transplantation. No significant changes in leukocyte adhesion or capillary blood flow were observed during the same time period. This report establishes multiphoton fluorescence microscopy as a sensitive tool for simultaneously studying functional and structural perturbations that occur in the mouse kidney after transplantation and for investigating the migration of leukocytes to the graft.