Surface imaging microscopy using an ultramiller for large volume 3D reconstruction of wax- and resin-embedded tissues
Surface imaging microscopy using an ultramiller for large volume 3D reconstruction of wax- and resin-embedded tissues
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DOI：
10.1002/jemt.20491
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发表时间：
2007-10-01
影响因子：
2.5
通讯作者：
Legrice, Ian J.
中科院分区：
文献类型：
--
作者：
Gerneke, Dane A.;Sands, Gregory B.;Legrice, Ian J.
使用组织学薄切片进行大组织体积的三维重建存在困难，因为切片的配准、切片失真以及由于切片丢失而可能收集不完整的数据集。我们已经构建了一个集成的表面成像系统，成功地解决了这些问题。嵌入的组织安装在高精度XYZ台上，上表面迭代：(i)染色以提供有效的光学切片，（ii）使用数码相机成像，（iii）用超细研磨机去除。这种方法通过固有地保持图像配准来重建高质量的图像，消除了截面失真，从而消除了复杂的重新排列和校正的需要，并确保了所有图像平面的完整捕获。该系统能够获取体素尺寸从0.5到50 μ m，尺寸从微米到几十毫米的组织结构图像。通过可靠地去除组织的全部范围，使大型样品能够成像。在如此大的尺度和分辨率范围内可视化3D组织结构的关键特征的能力将有助于更好地理解结构和功能之间的关系。这种理解对于更好地分析与不同疾病状态相关的结构变化以及开发基于结构的生物功能计算机模型至关重要。
Three-dimensional reconstruction of large tissue volumes using histological thin sections poses difficulties because of registration of sections, section distortion, and the possibility of incomplete data set collection due to section loss. We have constructed an integrated surface imaging system that successfully addresses these problems. Embedded tissue is mounted on a high precision XYZ stage and the upper surface is iteratively: (i) stained to provide an effective optical section, (ii) imaged using a digital camera, and (iii) removed with an ultramiller. This approach provides for the reconstruction of high-quality 31) images by inherently preserving image registration, eliminates section distortion, thus removing the need for complex realignment and correction, and also ensures full capture of all image planes. The system has the capacity to acquire images of tissue structure with voxel sizes from 0.5 to 50 mu m over dimensions ranging from micrometers to tens of millimeters. The ultramiller enables large samples to be imaged by reliably removing tissue over their full extent. The ability to visualize key features of 3D tissue structure across such a range of scale and resolution will facilitate the development of a greater understanding of the relationship between structure and function. This understanding is essential for better analyses of the structural changes associated with different disease states, and the development of structure-based computer models of biological function.