Optimized murine lung preparation for detailed structural evaluation via micro-computed tomography
Optimized murine lung preparation for detailed structural evaluation via micro-computed tomography
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DOI：
10.1152/japplphysiol.00550.2011
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发表时间：
2012-01-01
影响因子：
3.3
通讯作者：
Hoffman, Eric A.
中科院分区：
文献类型：
--
作者：
Vasilescu, Dragos M.;Knudsen, Lars;Hoffman, Eric A.
魏伯尔，何建平，何建平。基于微计算机断层扫描的小鼠肺组织结构评估。中国生物医学工程学报（英文版），2012；首次发表于2011年8月4日；doi: 10.1152 / japplphysiol.00550.2011。利用微x射线CT （mu CT）成像，我们试图从正常小鼠菌株中生成体内和完整/离体肺图谱。活体成像可以显示实质密度和小气道（15-28 μ m/体素）。通过mu CT对完整肺的离体成像可以提高对肺泡水平三维肺结构的理解，体素尺寸为1-2 μ m。mu CT要求空气空间保持充满以检测肺泡结构，同时保持肺的体内结构几何形状。为了达到这些要求，一种允许非破坏性全肺离体CT成像的固定和成像方法已经实施和测试。在体内成像后，在15、20和25 cmH(2)O气道压力下，将仰卧麻醉的C57Bl/6小鼠的肺通过血管灌注使用两阶段冲洗系统固定在原位，同时保持在20 cmH2O气道压力下。取出固定肺，风干。在肺干燥后1,7,21和bbb70天获得全肺体积，并用于固定稳定性验证。从体内到固定肺变化为+8.95% (P = 0.12)，从第1天到第7天变化为-1.47% (P = 0.07)，从第1天到第21天变化为-2.51% (P = 0.05)，从第1天到第70天及以后变化为-4.90% （P = 0.04）。CT检查显示肺泡和毛细血管床固定良好，组织学分析一致。建立了一种固定和成像方法，用于小鼠肺的mu CT成像，允许进行离体形态学分析，代表在体肺。
Vasilescu DM, Knudsen L, Ochs M, Weibel ER, Hoffman EA. Optimized murine lung preparation for detailed structural evaluation via micro-computed tomography. J Appl Physiol 112: 159-166, 2012. First published August 4, 2011; doi:10.1152/japplphysiol.00550.2011.Utilizing micro-X-ray CT (mu CT) imaging, we sought to generate an atlas of in vivo and intact/ex vivo lungs from normal murine strains. In vivo imaging allows visualization of parenchymal density and small airways (15-28 mu m/voxel). Ex vivo imaging of the intact lung via mu CT allows for improved understanding of the three-dimensional lung architecture at the alveolar level with voxel dimensions of 1-2 mu m. mu CT requires that air spaces remain air-filled to detect alveolar architecture while in vivo structural geometry of the lungs is maintained. To achieve these requirements, a fixation and imaging methodology that permits nondestructive whole lung ex vivo mu CT imaging has been implemented and tested. After in vivo imaging, lungs from supine anesthetized C57Bl/6 mice, at 15, 20, and 25 cmH(2)O airway pressure, were fixed in situ via vascular perfusion using a two-stage flushing system while held at 20 cmH2O airway pressure. Extracted fixed lungs were air-dried. Whole lung volume was acquired at 1, 7, 21, and >70 days after the lungs were dried and served as validation for fixation stability. No significant shrinkage was observed: +8.95% change from in vivo to fixed lung (P = 0.12), -1.47% change from day 1 to day 7 (P = 0.07), -2.51% change from day 1 to day 21 (P = 0.05), and -4.90% change from day 1 to day 70 and thereafter (P = 0.04). mu CT evaluation showed well-fixed alveoli and capillary beds correlating with histological analysis. A fixation and imaging method has been established for mu CT imaging of the murine lung that allows for ex vivo morphometric analysis, representative of the in vivo lung.