Synchrotron-based phase-contrast micro-CT as a tool for understanding pulmonary vascular pathobiology and the 3-D microanatomy of alveolar capillary dysplasia
Synchrotron-based phase-contrast micro-CT as a tool for understanding pulmonary vascular pathobiology and the 3-D microanatomy of alveolar capillary dysplasia
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DOI：
10.1152/ajplung.00103.2019
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发表时间：
2020-01-01
影响因子：
4.9
通讯作者：
Tran-Lundmark, Karin
中科院分区：
文献类型：
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作者：
Norvik, Christian;Westoo, Christian Karl;Tran-Lundmark, Karin
本研究旨在探讨同步加速器相位对比显微CT(Micro-CT)在肺血管病理生物学中的应用价值。肺的显微解剖结构复杂，有错综复杂的分支。因此，组织切片很难解释。肺内支气管肺吻合术(IBAS)已在几种形式的肺动脉高压中被描述，包括肺泡毛细血管发育不良伴肺静脉错位(ACD/MPV)。在这里，我们使用这种高分辨率三维成像的非破坏性方法检查石蜡包埋的组织。采用健康肺组织块和ACD/MPV肺组织块。ACD/MPV阻滞组的肺动脉和支气管动脉均预先注射了染料。在X02DA TOMCAT光束线(瑞士光源)的图像采集过程中，对每个块的一个切片进行染色，并标记感兴趣的区域，以实现精确的光束对准。X4放大镜与20微米厚的闪烁材料和sCMOS探测器相结合，在空间分辨率和视场之间取得了最佳的平衡。应用一种相位恢复算法，生成了成像体积的虚拟断层切片和视频剪辑。染料注射在血管和周围组织之间产生了明显的衰减差异，有利于分割和三维渲染。组织学和免疫组织化学成像后提供了互补的信息。ACD/MPV均为IBAS，MPV呈支气管状。我们展示了基于同步加速器的相位对比显微CT用于石蜡包埋组织中肺微血管解剖的三维表征的优势。血管染料注射增加了额外的价值。我们证实了ACD/MPV的肺内分流，并支持MPV是扩张的支气管静脉/小静脉的假设。
This study aimed to explore the value of synchrotron-based phase-contrast microcomputed tomography (micro-CT) in pulmonary vascular pathobiology. The microanatomy of the lung is complex with intricate branching patterns. Tissue sections are therefore difficult to interpret. Recruited intrapulmonary bronchopulmonary anastomoses (IBAs) have been described in several forms of pulmonary hypertension, including alveolar capillary dysplasia with misaligned pulmonary veins (ACD/MPV). Here, we examine paraffin-embedded tissue using this nondestructive method for high-resolution three-dimensional imaging. Blocks of healthy and ACD/MPV lung tissue were used. Pulmonary and bronchial arteries in the ACD/MPV block had been preinjected with dye. One section per block was stained, and areas of interest were marked to allow precise beam-alignment during image acquisition at the X02DA TOMCAT beamline (Swiss Light Source). A X4 magnifying objective coupled to a 20-mu m thick scintillating material and a sCMOS detector yielded the best trade-off between spatial resolution and field-of-view. A phase retrieval algorithm was applied and virtual tomographic slices and video clips of the imaged volumes were produced. Dye injections generated a distinct attenuation difference between vessels and surrounding tissue, facilitating segmentation and three-dimensional rendering. Histology and immunohistochemistry post-imaging offered complementary information. IBAs were confirmed in ACD/MPV, and the MPVs were positioned like bronchial veins/venules. We demonstrate the advantages of using synchrotron-based phase-contrast micro-CT for three-dimensional characterization of pulmonary microvascular anatomy in paraffin-embedded tissue. Vascular dye injections add additional value. We confirm intrapulmonary shunting in ACD/MPV and provide support for the hypothesis that MPVs are dilated bronchial veins/venules.