High Resolution Imaging of Mouse Embryos and Neonates with X-Ray Micro-Computed Tomography.

Q1 Agricultural and Biological Sciences

Current protocols in mouse biology Pub Date : 2019-06-01 Epub Date: 2019-06-13 DOI:10.1002/cpmo.63

Chih-Wei Hsu, Sowmya Kalaga, Uchechukwu Akoma, Tara L Rasmussen, Audrey E Christiansen, Mary E Dickinson

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引用次数: 9

Abstract

Iodine-contrast micro-computed tomography (microCT) 3D imaging provides a non-destructive and high-throughput platform for studying mouse embryo and neonate development. Here we provide protocols on preparing mouse embryos and neonates between embryonic day 8.5 (E8.5) to postnatal day 4 (P4) for iodine-contrast microCT imaging. With the implementation of the STABILITY method to create a polymer-tissue hybrid structure, we have demonstrated that not only is soft tissue shrinkage minimized but also the minimum required time for soft tissue staining with iodine is decreased, especially for E18.5 to P4 samples. In addition, we also provide a protocol on using commercially available X-CLARITY^TM hydrogel solution to create the similar polymer-tissue hybrid structure on delicate early post-implantation stage (E8.5 to E14.5) embryos. With its simple sample staining and mounting processes, this protocol is easy to adopt and implement for most of the commercially available, stand-alone microCT systems in order to study mouse development between early post-implantation to early postnatal stages. © 2019 by John Wiley & Sons, Inc.

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用x射线微计算机断层扫描技术对小鼠胚胎和新生儿进行高分辨率成像。

碘对比微计算机断层扫描(microCT)三维成像为研究小鼠胚胎和新生儿发育提供了一个非破坏性和高通量的平台。在这里，我们提供了制备小鼠胚胎和胚胎8.5天(E8.5)至出生后4天(P4)之间的新生儿用于碘造影剂微ct成像的方案。通过实施STABILITY方法来创建聚合物-组织杂化结构，我们已经证明，不仅软组织收缩最小化，而且软组织用碘染色所需的最短时间也减少了，特别是对于E18.5到P4样品。此外，我们还提供了一种使用市购X-CLARITYTM水凝胶溶液在胚胎植入后早期(E8.5至E14.5)建立类似聚合物-组织杂交结构的方案。由于其简单的样品染色和安装过程，该方案易于为大多数市售的独立微ct系统采用和实施，以便研究小鼠从植入后早期到产后早期阶段的发育。©2019 by John Wiley & Sons, Inc。

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来源期刊

Current protocols in mouse biology Agricultural and Biological Sciences-Animal Science and Zoology

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet, all too often, nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. The aim of Current Protocols in Mouse Biology is to provide the clearest, most detailed and reliable step-by-step instructions for protocols involving the use of mice in biomedical research. Written by experts in the field and extensively edited to our exacting standards, the protocols include all of the information necessary to complete an experiment in the laboratory—introduction, materials lists with supplier information, detailed step-by-step procedures with helpful annotations, recipes for reagents and solutions, illustrative figures and information-packed tables. Each article also provides invaluable discussions of background information, applications of the methods, important assumptions, key parameters, time considerations, and tips to help avoid common pitfalls and troubleshoot experiments. Furthermore, Current Protocols in Mouse Biology content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Mouse Biology brings together resources in mouse biology and genetics and provides a mouse protocol resource that covers all aspects of mouse biology. Current Protocols in Mouse Biology also permits optimization of mouse model usage, which is significantly impacted by both cost and ethical constraints. Optimal and standardized mouse protocols ultimately reduce experimental variability and reduce the number of animals used in mouse experiments.