In Vivo Fluorescence Imaging of Large-Vessel Thrombosis in Mice
In Vivo Fluorescence Imaging of Large-Vessel Thrombosis in Mice
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DOI：
10.1161/atvbaha.111.225334
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发表时间：
2011-06-01
影响因子：
8.7
通讯作者：
Cooley, Brian C.
中科院分区：
文献类型：
--
作者：
Cooley, Brian C.
大血管血栓形成的客观实验研究已适用于小鼠，但它们在结果测量中提供的可量化信息有限。本研究提出了一种通过时间/空间可量化测量和动物间比较归一化方法来评估大血管血栓形成的新方法。方法和结果-快门式扩束激光器为具有大焦深的 x100 显微手术野提供均匀的窄波长照明。通过短暂的血管表面电解损伤，在小鼠颈动脉和股静脉中产生血栓。血栓靶向荧光团被全身注射，随后定位在血栓诱导部位。带有滤光轮的低光数码摄像机可在 60 分钟的时间间隔内采集特定目标的图像。血小板积聚，随后出现纤维蛋白边界，以稳定动脉和静脉中的凝块。凝血酶复合物与纤维蛋白沉积共定位。大动脉经历周期性大量血栓栓塞，而静脉则显示微栓子逐渐脱落和血栓收缩。全身施用纤维蛋白和血小板抑制化合物可减少其各自的靶标，但也常常抑制动脉和静脉中的凝血对应物（分别为血小板和纤维蛋白）。结论-中级放大图像捕获代表了一种基于荧光的体内成像分析的新方法，可定量应用于大血管血栓形成的研究。 （动脉硬化血栓 Vasc Biol.2011；31：1351-1356。）
Objective-Experimental studies of large-vessel thrombosis have been adapted for applications in mice, but they proffer limited quantifiable information in outcome measures. This study presents a novel approach for evaluating large-vessel thrombogenesis with temporally/spatially quantifiable measures and normalization methods for interanimal comparisons.Methods and Results-Shuttered, beam-expanded lasers provided uniform narrow-wavelength illumination of a x100 microsurgical field with a large depth of focus. Thrombosis was generated in murine carotid arteries and femoral veins by brief vascular surface electrolytic injury. Thrombus-targeting fluorophores were injected systemically and subsequently localized at the site of thrombus induction. A low-light digital video camera with filter wheel provided target-specific image acquisition over a 60-minute interval. Platelets accumulated with a subsequent fibrin border emerging to stabilize the clot in both arteries and veins. Coagulation enzyme complexes colocalized with fibrin deposition. Large arteries underwent cyclic massive thromboembolization, whereas veins showed gradual shedding of microemboli and clot contraction. Systemic administration of fibrin-and platelet-inhibiting compounds reduced their respective targets but also often inhibited their clotting counterparts (platelets and fibrin, respectively) in both arteries and veins.Conclusion-Intermediate-level magnified image capture represents a novel approach for analysis of fluorescence-based in vivo imaging, with quantitative application to the study of large-vessel thrombosis. (Arterioscler Thromb Vasc Biol. 2011;31:1351-1356.)