3D cryo-imaging: a very high-resolution view of the whole mouse.
3D cryo-imaging: a very high-resolution view of the whole mouse.
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DOI：
10.1002/ar.20849
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发表时间：
2009-03
影响因子：
2
通讯作者：
Wilson, David L.
中科院分区：
文献类型：
--
作者：
Roy, Debashish;Steyer, Grant J.;Gargesha, Madhusudhana;Stone, Meredith E.;Wilson, David L.
我们开发了CASE冷冻成像系统，它使用切片和图像块脸成像技术提供信息丰富的、非常高分辨率的彩色亮场图像和整个小鼠的分子荧光图像。该系统包括一个鼠标大小的、具有特殊成像功能的电动冷冻切割机、一个改装的Brightfield/荧光显微镜和一个机器人XYZ成像系统定位器，所有这些都是由控制系统完全自动化的。使用机器人系统，我们在一只40微米切片的整个鼠标的块面上采集了15.6微米像素大小的显微平铺图像，总数据量为55 GB。通过观察不同分辨率的2D图像，我们发现了心脏血管、肌层、小肠绒毛、视神经和眼部各层等小结构。冷冻成像也适用于胚胎突变的三维成像。在一只小鼠中，在伽马肌动蛋白启动子的作用下，增强的绿色荧光蛋白在平滑肌细胞中表达，从而提供了清晰的泌尿生殖和胃肠道平滑肌的3D图像。通过冷冻成像，我们可以在小鼠大脑的非常大范围内获得低至10微米的3D血管系统。软件完全自动化，具有完全可编程的成像/切片协议、电子邮件通知和自动体积可视化。凭借独特的视野、景深、对比度和分辨率的组合，CASE冷冻成像系统填补了整个动物活体成像和组织学之间的空白。
We developed the Case Cryo-imaging system that provides information rich, very high-resolution, color brightfield, and molecular fluorescence images of a whole mouse using a section-and-image block-face imaging technology. The system consists of a mouse-sized, motorized cryo-microtome with special features for imaging, a modified, brightfield/ fluorescence microscope, and a robotic xyz imaging system positioner, all of which is fully automated by a control system. Using the robotic system, we acquired microscopic tiled images at a pixel size of 15.6 µm over the block face of a whole mouse sectioned at 40 µm, with a total data volume of 55 GB. Viewing 2D images at multiple resolutions, we identified small structures such as cardiac vessels, muscle layers, villi of the small intestine, the optic nerve, and layers of the eye. Cryo-imaging was also suitable for imaging embryo mutants in 3D. A mouse, in which enhanced green fluorescent protein was expressed under gamma actin promoter in smooth muscle cells, gave clear 3D views of smooth muscle in the urogenital and gastrointestinal tracts. With cryo-imaging, we could obtain 3D vasculature down to 10 µm, over very large regions of mouse brain. Software is fully automated with fully programmable imaging/sectioning protocols, email notifications, and automatic volume visualization. With a unique combination of field-of-view, depth of field, contrast, and resolution, the Case Cryo-imaging system fills the gap between whole animal in vivo imaging and histology.
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影响因子：
30.8
作者：
Bamforth, SD;Bragança, J;Bhattacharya, S
通讯作者：
Bhattacharya, S
影响因子：
4.5
作者：
Brown SD;Hancock JM;Gates H
通讯作者：
Gates H
影响因子：
5.1
作者：
MacLaren, DC;Gambhir, SS;Herschman, HR
通讯作者：
Herschman, HR
影响因子：
4.8
作者：
Lazebnik, RS;Weinberg, BD;Wilson, DL
通讯作者：
Wilson, DL
影响因子：
2
作者：
Griffini, P;Smorenburg, SM;VanNoorden, CJF
通讯作者：
VanNoorden, CJF