Gregg Chelsea L, Recknagel Andrew K, Butcher Jonathan T
Department of Biomedical Engineering, Cornell University, 304 Weill Hall, Ithaca, NY, USA.
Methods Mol Biol. 2015;1189:47-61. doi: 10.1007/978-1-4939-1164-6_4.
Tissue morphogenesis and embryonic development are dynamic events challenging to quantify, especially considering the intricate events that happen simultaneously in different locations and time. Micro- and more recently nano-computed tomography (micro/nanoCT) has been used for the past 15 years to characterize large 3D fields of tortuous geometries at high spatial resolution. We and others have advanced micro/nanoCT imaging strategies for quantifying tissue- and organ-level fate changes throughout morphogenesis. Exogenous soft tissue contrast media enables visualization of vascular lumens and tissues via extravasation. Furthermore, the emergence of antigen-specific tissue contrast enables direct quantitative visualization of protein and mRNA expression. Micro-CT X-ray doses appear to be non-embryotoxic, enabling longitudinal imaging studies in live embryos. In this chapter we present established soft tissue contrast protocols for obtaining high-quality micro/nanoCT images and the image processing techniques useful for quantifying anatomical and physiological information from the data sets.
组织形态发生和胚胎发育是动态事件，难以量化，尤其是考虑到在不同位置和时间同时发生的复杂事件。在过去15年中，微型以及最近的纳米计算机断层扫描（微/纳米CT）已被用于以高空间分辨率表征具有复杂几何形状的大型三维区域。我们和其他人已经改进了微/纳米CT成像策略，以量化整个形态发生过程中组织和器官水平的命运变化。外源性软组织造影剂能够通过渗漏使血管腔和组织可视化。此外，抗原特异性组织对比的出现使得能够直接定量可视化蛋白质和mRNA表达。微CT X射线剂量似乎无胚胎毒性，从而能够对活胚胎进行纵向成像研究。在本章中，我们介绍了用于获取高质量微/纳米CT图像的既定软组织对比方案以及有助于从数据集中量化解剖和生理信息的图像处理技术。
