Group of CNS Angiogenesis and Neurovascular Link, Neuroscience Center Zurich, and Division of Neurosurgery, University and University Hospital Zurich, Zurich, Switzerland.
Division of Neurosurgery, University Hospital Zurich, Zurich, Switzerland.
Nat Protoc. 2021 Oct;16(10):4564-4610. doi: 10.1038/s41596-021-00587-1. Epub 2021 Sep 3.
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.
在大脑发育过程中、在成年健康大脑中以及在中枢神经系统的各种疾病中，新血管的形成和血管网络的建立至关重要。在这里，我们描述了我们最近开发的一种方法的分步协议，该方法能够对新生和成年小鼠大脑中的血管网络进行层次成像和计算分析。该程序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、同步辐射和台式微计算机断层扫描成像以及计算网络分析。将这些方法结合起来，可以详细可视化和量化 3D 脑血管。可以在从新生早期到成年大脑的任何发育阶段获得网络特征，如血管体积分数、分支点密度、血管直径、长度、迂曲度和方向性以及血管外距离。这种方法可用于提供整个小鼠脑血管系统在新生和成年发育阶段的详细形态图谱。我们的方案允许分别对毛细血管和非毛细血管的脑血管网络进行特征描述。从小鼠灌注到血管网络分析，整个方案大约需要 10 天。
