Three-dimensional virtual histology of human cerebellum by X-ray phase-contrast tomography.
Three-dimensional virtual histology of human cerebellum by X-ray phase-contrast tomography.
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DOI：
10.1073/pnas.1801678115
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发表时间：
2018-07-03
影响因子：
11.1
通讯作者：
Salditt T
中科院分区：
文献类型：
--
作者：
Töpperwien M;van der Meer F;Stadelmann C;Salditt T
人脑组织的复杂细胞结构传统上由组织学研究，在2D平面上提供结构信息。然而，通过以非各向同性分辨率检查许多平行切片，这可以部分扩展到3D。这项工作表明，基于传播的X射线相衬断层成像，无论是在同步加速器上，还是在紧凑的实验室源上，都可以用于在各向同性亚细胞分辨率下对未染色的石蜡包埋的人小脑进行非侵入性的3D虚拟组织学研究。由此产生的数据质量足以可视化并自动定位小脑不同层中的∼106神经元，提供了关于其3D细胞结构和空间组织的前所未有的数据。为了定量评估脑组织及其相应的功能，三维细胞分布的知识是必不可少的。获得这些信息的黄金标准是组织学，这是一种破坏性的劳动密集型技术，将标本切片并在光学显微镜下进行检查，以非各向同性分辨率提供3D信息。为了克服传统组织学的局限性，我们在配备了用于相干和波前滤波的X射线波导光学元件的专用同步辐射终端站和紧凑的实验室光源上，使用具有优化光学、重建和图像分析的相衬X射线断层扫描。作为概念验证的演示，我们探索了石蜡包埋的未染色的人类小脑的毫米大小的3D细胞结构，并表明在整个标本的两种设置下都可以达到各向同性的亚细胞分辨率。为了能够对重建的数据进行定量分析，我们演示了小脑皮质内100多万个神经元的自动细胞分割和定位。这使得可以通过借用凝聚态物理的概念来分析所有维度上细胞的空间组织和相关性，这表明了颗粒层中细胞的强烈短程有序和局部聚集。通过对3D神经元“堆积”的量化，我们可以清楚地了解人类小脑是如何以其体积的10%容纳大脑中80%的神经元的。此外，我们还证明了颗粒层中相邻神经元的分布相对于浦肯野细胞树突是各向异性的。
The complex cytoarchitecture of human brain tissue is traditionally studied by histology, providing structural information in 2D planes. This can be partly extended to 3D by inspecting many parallel slices, however, at nonisotropic resolution. This work shows that propagation-based X-ray phase-contrast tomography, both at the synchrotron and even at a compact laboratory source, can be used to perform noninvasive 3D virtual histology on unstained paraffin-embedded human cerebellum at isotropic subcellular resolution. The resulting data quality is high enough to visualize and automatically locate ∼106 neurons within the different layers of the cerebellum, providing unprecedented data on its 3D cytoarchitecture and spatial organization. To quantitatively evaluate brain tissue and its corresponding function, knowledge of the 3D cellular distribution is essential. The gold standard to obtain this information is histology, a destructive and labor-intensive technique where the specimen is sliced and examined under a light microscope, providing 3D information at nonisotropic resolution. To overcome the limitations of conventional histology, we use phase-contrast X-ray tomography with optimized optics, reconstruction, and image analysis, both at a dedicated synchrotron radiation endstation, which we have equipped with X-ray waveguide optics for coherence and wavefront filtering, and at a compact laboratory source. As a proof-of-concept demonstration we probe the 3D cytoarchitecture in millimeter-sized punches of unstained human cerebellum embedded in paraffin and show that isotropic subcellular resolution can be reached at both setups throughout the specimen. To enable a quantitative analysis of the reconstructed data, we demonstrate automatic cell segmentation and localization of over 1 million neurons within the cerebellar cortex. This allows for the analysis of the spatial organization and correlation of cells in all dimensions by borrowing concepts from condensed-matter physics, indicating a strong short-range order and local clustering of the cells in the granular layer. By quantification of 3D neuronal “packing,” we can hence shed light on how the human cerebellum accommodates 80% of the total neurons in the brain in only 10% of its volume. In addition, we show that the distribution of neighboring neurons in the granular layer is anisotropic with respect to the Purkinje cell dendrites.
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DOI：
10.1364/josaa.26.000890
发表时间：
2009-04-01
影响因子：
1.9
作者：
De Witte, Yoni;Boone, Matthieu;Van Hoorebeke, Luc
通讯作者：
Van Hoorebeke, Luc
影响因子：
3.7
作者：
Jiao, Yang;Berman, Hal;Torquato, Salvatore
通讯作者：
Torquato, Salvatore
DOI：
10.1073/pnas.1204460109
发表时间：
2012-11-06
影响因子：
11.1
作者：
Zhao, Yunzhe;Brun, Emmanuel;Bravin, Alberto
通讯作者：
Bravin, Alberto
影响因子：
1.6
作者：
Otendal, M.;Tuohimaa, T.;Hertz, H. M.
通讯作者：
Hertz, H. M.
影响因子：
1.6
作者：
Zabler, S;Cloetens, P;Schlenker, M
通讯作者：
Schlenker, M