Large Scale Imaging by Fine Spatial Alignment of Multi-Scanning Data with Gel Cube Device
Large Scale Imaging by Fine Spatial Alignment of Multi-Scanning Data with Gel Cube Device
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DOI：
10.3390/app8020235
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发表时间：
2018-02-01
影响因子：
2.7
通讯作者：
Kawahara, Tomohiro
中科院分区：
文献类型：
--
作者：
Hagiwara, Masaya;Nobata, Rina;Kawahara, Tomohiro
体外三维（3D）培养被认为在许多生物学领域中是必不可少的。然而，开发的3D地层的成像通常是困难的，特别是如果样品的尺寸相对较大。由于大的焦深，使用低放大率透镜（4x和10x）的荧光成像的z分辨率低。本文描述了3D培养平台，通过对从多个方向获得的图像数据集进行精细的空间对准来实现大规模3D成像。采用凝胶立方体装置进行多次扫描，然后在立方体框架中的自荧光微结构允许我们在几个像素内空间对齐图像数据集。通过合成来自多个扫描的数据，该平台使我们能够同时可视化毫米大小的3D样品结构和单个细胞肌动蛋白丝。毫米级深度成像的支气管树实现了高z分辨率。该设备，这是适用于大多数显微镜系统，可以提高图像质量，而无需修改当前的系统。
In vitro three-dimensional (3D) culturing is considered essential in many biological fields. However, the imaging of developed 3D formations is often difficult, especially if the size of the sample is relatively large. The z-resolution of fluorescent imaging is low using low magnification lenses (4x and 10x) due to large focal depths. This paper describes 3D culture platform enabling large scale 3D imaging by fine spatial alignment of the image dataset obtained from multiple directions. A gel cube device was employed to conduct the multi-scanning and then a self-fluorescent microstructure in a cubic frame allows us spatially align image dataset within a few pixels. By synthesizing data from multiple scans, the platform enables us to visualize millimeter-sized 3D sample structure and individual cellular actin filaments at the same time. Millimeter depth imaging of a developed bronchial tree was achieved with high z-resolution. The device, which is applicable to most microscopy systems, can enhance the image quality without modifying current systems.