Phosphotungstic acid-enhanced microCT: Optimized protocols for embryonic and early postnatal mice
Phosphotungstic acid-enhanced microCT: Optimized protocols for embryonic and early postnatal mice
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DOI：
10.1002/dvdy.136
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发表时间：
2019-11-28
影响因子：
2.5
通讯作者：
Richtsmeier, Joan T.
中科院分区：
文献类型：
--
作者：
Lesciotto, Kate M.;Perrine, Susan M. Motch;Richtsmeier, Joan T.
背景由于需要描述性的和日益机械化的形态分析，对比度增强的显微计算机断层扫描(MicroCT)可能是原位显示三维生物软组织的最好方法。尽管使用磷钨酸(PTA)的染色方案已经发表，提供了漂亮的软组织结构可视化，但这些方案往往针对高度特定的研究问题，并适用于有限的一组模型生物、标本年龄或组织类型。我们提供了使用PTA增强的MicroCT对几个胚胎和出生后早期的小鼠的软组织结构进行微观水平可视化的详细方案。结果我们的方案产生了MicroCT扫描，使整个生物体、单个组织和器官系统能够可视化和定量分析，同时保留3D形态和与周围结构的关系，软组织收缩最小。特别值得注意的是，以高分辨率捕捉到了小鼠心脏、肺和肝脏的内部和外部特征，以及胚胎软骨。结论这些方案对各种疾病和条件下的小鼠模型具有广泛的适用性。在染色持续时间内进行较小的实验可以将该方案扩展到其他年龄段，从而允许在其3D原位位置内对内部器官和软组织结构进行个体发育研究。
Background Given the need for descriptive and increasingly mechanistic morphological analyses, contrast-enhanced microcomputed tomography (microCT) represents perhaps the best method for visualizing 3D biological soft tissues in situ. Although staining protocols using phosphotungstic acid (PTA) have been published with beautiful visualizations of soft tissue structures, these protocols are often aimed at highly specific research questions and are applicable to a limited set of model organisms, specimen ages, or tissue types. We provide detailed protocols for micro-level visualization of soft tissue structures in mice at several embryonic and early postnatal ages using PTA-enhanced microCT. Results Our protocols produce microCT scans that enable visualization and quantitative analyses of whole organisms, individual tissues, and organ systems while preserving 3D morphology and relationships with surrounding structures, with minimal soft tissue shrinkage. Of particular note, both internal and external features of the murine heart, lungs, and liver, as well as embryonic cartilage, are captured at high resolution. Conclusion These protocols have broad applicability to mouse models for a variety of diseases and conditions. Minor experimentation in the staining duration can expand this protocol to additional age groups, permitting ontogenetic studies of internal organs and soft tissue structures within their 3D in situ position.