Coronary artery wall imaging in mice using osmium tetroxide and micro-computed tomography (micro-CT).
Coronary artery wall imaging in mice using osmium tetroxide and micro-computed tomography (micro-CT).
复制标题
使用四氧化锇和微型计算机断层扫描 (micro-CT) 对小鼠进行冠状动脉壁成像。
DOI：
10.1111/j.1469-7580.2012.01483.x
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发表时间：
2012-05
影响因子：
2.4
通讯作者：
Wen H
中科院分区：
文献类型：
--
作者：
Pai VM;Kozlowski M;Donahue D;Miller E;Xiao X;Chen MY;Yu ZX;Connelly P;Jeffries K;Wen H
微型计算机断层扫描（micro-CT）的高空间分辨率是完整小鼠心脏标本冠状动脉三维成像的理想选择。以前，小鼠心脏标本的微CT利用血管内造影剂，其在血管腔内硬化并允许制作血管铸型。然而，对于小鼠冠状动脉疾病模型，非常希望对冠状动脉壁进行成像并突出斑块。为了这个目的，我们描述了一种离体对比度增强显微CT成像技术的基础上与四氧化锇（OsO 4）溶液的组织染色。作为组织染色造影剂，OsO 4在固定过程中保留在血管壁和周围组织中，并从血管腔中清除。其高X射线衰减使得动脉壁在CT中可见。此外，由于OsO 4优先与脂质结合，它突出了动脉壁中的脂质沉积。我们对5- 25周龄C57 BL/6野生型小鼠和5- 13周龄载脂蛋白E基因敲除（apoE -/-）小鼠的心脏标本进行了显微CT，分辨率为10 μm。结果表明，使用台式微型CT扫描仪可以看到直径小至45 μm的冠状动脉壁。通过使用同步加速器CT扫描仪，以1/2000的扫描时间实现了类似的图像清晰度。在13周龄的apoE小鼠中，主动脉中可见富含脂质的斑块。我们的研究表明，OsO 4和micro-CT的组合允许在完整的小鼠心脏冠状动脉壁的可视化。
The high spatial resolution of Micro-Computed Tomography (micro-CT) is ideal for 3D imaging of coronary arteries in intact mouse heart specimens. Previously, micro-CT of mouse heart specimens utilized intravascular contrast agents that hardened within the vessel lumen and allowed a vascular cast to be made. However, for mouse coronary artery disease models, it is highly desirable to image coronary artery walls and highlight plaques. For this purpose, we describe an ex vivo contrast-enhanced micro-CT imaging technique based on tissue staining with osmium tetroxide (OsO4) solution. As a tissue-staining contrast agent, OsO4 is retained in the vessel wall and surrounding tissue during the fixation process and cleared from the vessel lumens. Its high X-ray attenuation makes the artery wall visible in CT. Additionally, since OsO4 preferentially binds to lipids, it highlights lipid deposition in the artery wall. We performed micro-CT of heart specimens of 5-to 25-week old C57BL/6 wild-type mice and 5-to13-week old apolipoprotein E knockout (apoE -/-) mice at 10 μm resolution. The results show that walls of coronary arteries as small as 45 μm in diameter are visible using a table-top micro-CT scanner. Similar image clarity was achieved with 1/2000th the scan time by using a synchrotron CT scanner. In 13-week old apoE mice, lipid-rich plaques are visible in the aorta. Our study shows that the combination of OsO4 and micro-CT permit the visualization of the coronary artery wall in intact mouse hearts.
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