Optical clearing and fluorescence deep-tissue imaging for 3D quantitative analysis of the brain tumor microenvironment.
Optical clearing and fluorescence deep-tissue imaging for 3D quantitative analysis of the brain tumor microenvironment.
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DOI：
10.1007/s10456-017-9565-6
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发表时间：
2017-11
期刊：
影响因子：
9.8
通讯作者：
Wurdinger T
中科院分区：
文献类型：
--
作者：
Lagerweij T;Dusoswa SA;Negrean A;Hendrikx EML;de Vries HE;Kole J;Garcia-Vallejo JJ;Mansvelder HD;Vandertop WP;Noske DP;Tannous BA;Musters RJP;van Kooyk Y;Wesseling P;Zhao XW;Wurdinger T
脑血管系统及其与周围细胞相互作用的三维可视化可能有助于了解涉及异常微血管组织的疾病，包括胶质母细胞瘤（GBM）。活体共聚焦成像允许小鼠脑中微血管结构和细胞迁移的3D可视化，然而，成像深度有限。为了能够全面分析GBM和大脑微环境，需要深入的3D成像方法。在这里，我们采用的方法，光学组织清除之前，三维显微镜可视化脑微血管和GBM细胞的入侵途径。我们提出了一个工作流程，光学透明的脑肿瘤组织的离体成像和随后的计算建模。该工作流程用于定量健康小鼠脑组织和人原位浸润性GBM 8和E98胶质母细胞瘤模型中与核或细胞密度相关的微血管。 与体内小鼠脑的活体成像相比，离体清除的小鼠脑组织具有>10倍的成像深度。光学透明的脑组织成像允许量化健康小鼠脑和具有弥漫性、浸润性生长的GBM 8脑肿瘤的组织中的3D微血管特征。详细的3D可视化显示了灰质和白色物质区域中肿瘤细胞相对于脉管系统的组织，以及集体侵入脑实质的多细胞GBM网络的模式。光学组织清除为GBM细胞及其微环境之间的地形关系的组合定量和3D显微镜分析开辟了新的途径。本文的在线版本（doi：10.1007/s10456-017-9565-6）包含补充材料，可供授权用户使用。
Three-dimensional visualization of the brain vasculature and its interactions with surrounding cells may shed light on diseases where aberrant microvascular organization is involved, including glioblastoma (GBM). Intravital confocal imaging allows 3D visualization of microvascular structures and migration of cells in the brain of mice, however, with limited imaging depth. To enable comprehensive analysis of GBM and the brain microenvironment, in-depth 3D imaging methods are needed. Here, we employed methods for optical tissue clearing prior to 3D microscopy to visualize the brain microvasculature and routes of invasion of GBM cells. We present a workflow for ex vivo imaging of optically cleared brain tumor tissues and subsequent computational modeling. This workflow was used for quantification of the microvasculature in relation to nuclear or cellular density in healthy mouse brain tissues and in human orthotopic, infiltrative GBM8 and E98 glioblastoma models. Ex vivo cleared mouse brain tissues had a >10-fold imaging depth as compared to intravital imaging of mouse brain in vivo. Imaging of optically cleared brain tissue allowed quantification of the 3D microvascular characteristics in healthy mouse brains and in tissues with diffuse, infiltrative growing GBM8 brain tumors. Detailed 3D visualization revealed the organization of tumor cells relative to the vasculature, in both gray matter and white matter regions, and patterns of multicellular GBM networks collectively invading the brain parenchyma. Optical tissue clearing opens new avenues for combined quantitative and 3D microscopic analysis of the topographical relationship between GBM cells and their microenvironment. The online version of this article (doi:10.1007/s10456-017-9565-6) contains supplementary material, which is available to authorized users.
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影响因子：
12.7
作者：
Caretti V;Sewing AC;Lagerweij T;Schellen P;Bugiani M;Jansen MH;van Vuurden DG;Navis AC;Horsman I;Vandertop WP;Noske DP;Wesseling P;Kaspers GJ;Nazarian J;Vogel H;Hulleman E;Monje M;Wurdinger T
通讯作者：
Wurdinger T
影响因子：
3
作者：
Bjornsson, Christopher S.;Lin, Gang;Roysam, Badrinath
通讯作者：
Roysam, Badrinath
影响因子：
48
作者：
Chung, Kwanghun;Deisseroth, Karl
通讯作者：
Deisseroth, Karl
影响因子：
9.8
作者：
Gilhuis, H. Jacobus;van der Laak, Jeroen A. W. M.;Wesseling, Pieter
通讯作者：
Wesseling, Pieter
影响因子：
2.3
作者：
Jaehrling, Nina;Becker, Klaus;Dodt, Hans-Ulrich
通讯作者：
Dodt, Hans-Ulrich