Micheva Kristina D, O'Rourke Nancy, Busse Brad, Smith Stephen J
Cold Spring Harb Protoc. 2010 Nov 1;2010(11):pdb.prot5527. doi: 10.1101/pdb.prot5527.
Array tomography is a volumetric microscopy method based on physical serial sectioning. Ultrathin sections of a plastic-embedded tissue are cut using an ultramicrotome, bonded in an ordered array to a glass coverslip, stained as desired, and imaged. The resulting two-dimensional image tiles can then be reconstructed computationally into three-dimensional volume images for visualization and quantitative analysis. The minimal thickness of individual sections permits high-quality rapid staining and imaging, whereas the array format allows reliable and convenient section handling, staining, and automated imaging. Also, the physical stability of the arrays permits images to be acquired and registered from repeated cycles of staining, imaging, and stain elution, as well as from imaging using multiple modalities (e.g., fluorescence and electron microscopy). Array tomography makes it possible to visualize and quantify previously inaccessible features of tissue structure and molecular architecture. However, careful preparation of the tissue is essential for successful array tomography; these steps can be time-consuming and require some practice to perfect. Successful array tomography requires that the captured images be properly stacked and aligned, and the software to achieve these ends is freely available. This protocol describes the construction of volumetric image stacks from images of fluorescently labeled arrays for three-dimensional image visualization, analysis, and archiving.
阵列断层扫描是一种基于物理连续切片的体视显微镜方法。使用超薄切片机切割塑料包埋组织的超薄切片，将其按有序阵列粘结到玻璃盖玻片上，根据需要进行染色并成像。然后可以通过计算机将所得的二维图像切片重建为三维体积图像，用于可视化和定量分析。单个切片的最小厚度允许进行高质量的快速染色和成像，而阵列形式则便于可靠且方便地处理切片、染色和自动成像。此外，阵列的物理稳定性使得能够从染色、成像和洗脱染色剂的重复循环以及使用多种模式（例如荧光和电子显微镜）成像中采集和配准图像。阵列断层扫描使得可视化和量化以前无法观察到的组织结构和分子结构特征成为可能。然而，仔细制备组织对于成功进行阵列断层扫描至关重要；这些步骤可能很耗时，并且需要一些实践才能完善。成功的阵列断层扫描要求捕获的图像正确堆叠和对齐，并且有免费的软件可用于实现这些目的。本方案描述了如何从荧光标记阵列的图像构建体积图像堆栈，用于三维图像可视化、分析和存档。
