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Hierarchical imaging and computational analysis of three-dimensional vascular network architecture in the entire postnatal and adult mouse brain
Nature Protocols ( IF 14.8 ) Pub Date : 2021-09-03 , DOI: 10.1038/s41596-021-00587-1
Thomas Wälchli 1, 2, 3, 4 , Jeroen Bisschop 1, 2, 3, 4, 5 , Arttu Miettinen 6, 7, 8 , Alexandra Ulmann-Schuler 9 , Christoph Hintermüller 10 , Eric P Meyer 9 , Thomas Krucker 11 , Regula Wälchli 12 , Philippe P Monnier 5, 13, 14 , Peter Carmeliet 15, 16 , Johannes Vogel 17 , Marco Stampanoni 6, 7
Affiliation  

The formation of new blood vessels and the establishment of vascular networks are crucial during brain development, in the adult healthy brain, as well as in various diseases of the central nervous system. Here, we describe a step-by-step protocol for our recently developed method that enables hierarchical imaging and computational analysis of vascular networks in postnatal and adult mouse brains. The different stages of the procedure include resin-based vascular corrosion casting, scanning electron microscopy, synchrotron radiation and desktop microcomputed tomography imaging, and computational network analysis. Combining these methods enables detailed visualization and quantification of the 3D brain vasculature. Network features such as vascular volume fraction, branch point density, vessel diameter, length, tortuosity and directionality as well as extravascular distance can be obtained at any developmental stage from the early postnatal to the adult brain. This approach can be used to provide a detailed morphological atlas of the entire mouse brain vasculature at both the postnatal and the adult stage of development. Our protocol allows the characterization of brain vascular networks separately for capillaries and noncapillaries. The entire protocol, from mouse perfusion to vessel network analysis, takes ~10 d.



中文翻译:

整个产后和成年小鼠大脑三维血管网络结构的分层成像和计算分析

新血管的形成和血管网络的建立在大脑发育、成人健康大脑以及中枢神经系统的各种疾病中都是至关重要的。在这里,我们描述了我们最近开发的方法的分步协议,该方法可以对产后和成年小鼠大脑中的血管网络进行分层成像和计算分析。该过程的不同阶段包括基于树脂的血管腐蚀铸造、扫描电子显微镜、同步辐射和桌面微型计算机断层扫描成像以及计算网络分析。结合这些方法可以实现 3D 脑血管系统的详细可视化和量化。血管体积分数、分支点密度、血管直径、长度等网络特征,从出生后早期到成年大脑的任何发育阶段都可以获得曲折性和方向性以及血管外距离。这种方法可用于在出生后和成年发育阶段提供整个小鼠大脑血管的详细形态学图谱。我们的协议允许分别为毛细血管和非毛细血管表征脑血管网络。整个协议,从鼠标灌注到血管网络分析,需要大约 10 天。我们的协议允许分别为毛细血管和非毛细血管表征脑血管网络。整个协议,从鼠标灌注到血管网络分析,需要大约 10 天。我们的协议允许分别为毛细血管和非毛细血管表征脑血管网络。整个协议,从鼠标灌注到血管网络分析,需要大约 10 天。

更新日期:2021-09-03
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