Morphological Characterisation of Unstained and Intact Tissue Micro-architecture by X-ray Computed Micro- and Nano-Tomography.
Morphological Characterisation of Unstained and Intact Tissue Micro-architecture by X-ray Computed Micro- and Nano-Tomography.
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DOI：
10.1038/srep10074
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发表时间：
2015-05-15
影响因子：
4.6
通讯作者：
Sherratt MJ
中科院分区：
文献类型：
--
作者：
Walton LA;Bradley RS;Withers PJ;Newton VL;Watson RE;Austin C;Sherratt MJ
组织结构的表征和量化受到切片诱导的伪影以及可视化和分割3D体积的困难的限制。在这里，我们证明，即使在没有X射线造影剂的情况下，X射线计算机显微断层扫描（microCT）和纳米断层扫描（nanoCT）也可以通过快速解析成分离散的3D组织区域（例如富含胶原蛋白的外膜和富含弹性蛋白的外膜）来规避这些问题完整的大鼠动脉中的薄片），而这些区域又可以由于其不同的X射线不透明度和形态而被分割。然后，我们建立，使用未加压和加压动脉的X射线断层扫描，腔内压力不仅增加管腔横截面积和拉直内侧弹性层，而且还诱导外膜层的深刻重塑。最后，我们将microCT应用于另一个人体器官（皮肤），以可视化细胞丰富的表皮和细胞外基质丰富的真皮，并显示传统的组织学和免疫组织化学染色方案与先前的X射线曝光兼容。因此，我们建议microCT可以结合光学显微镜，以确定档案石蜡包埋的生物材料和机械应力的动态组织，如心脏，肺和肌腱的三维结构和组成。
Characterisation and quantification of tissue structures is limited by sectioning-induced artefacts and by the difficulties of visualising and segmenting 3D volumes. Here we demonstrate that, even in the absence of X-ray contrast agents, X-ray computed microtomography (microCT) and nanotomography (nanoCT) can circumvent these problems by rapidly resolving compositionally discrete 3D tissue regions (such as the collagen-rich adventitia and elastin-rich lamellae in intact rat arteries) which in turn can be segmented due to their different X-ray opacities and morphologies. We then establish, using X-ray tomograms of both unpressurised and pressurised arteries that intra-luminal pressure not only increases lumen cross-sectional area and straightens medial elastic lamellae but also induces profound remodelling of the adventitial layer. Finally we apply microCT to another human organ (skin) to visualise the cell-rich epidermis and extracellular matrix-rich dermis and to show that conventional histological and immunohistochemical staining protocols are compatible with prior X-ray exposure. As a consequence we suggest that microCT could be combined with optical microscopy to characterise the 3D structure and composition of archival paraffin embedded biological materials and of mechanically stressed dynamic tissues such as the heart, lungs and tendons.