Much of our understanding of the spatial organization of and interactions between cellular organelles and macromolecular complexes has been the result of imaging studies utilizing either light- or electron-based microscopic analyses. These classical approaches, while insightful, are nonetheless limited either by restrictions in resolution or by the sheer complexity of generating multidimensional data. Recent advances in the use and application of X-rays to acquire micro- and nanotomographic data sets offer an alternative methodology to visualize cellular architecture at the nanoscale. These new approaches allow for the subcellular analyses of unstained vitrified cells and three-dimensional localization of specific protein targets and have served as an essential tool in bridging light and electron correlative microscopy experiments. Here, we review the theory, instrumentation details, acquisition principles, and applications of both soft X-ray tomography and X-ray microscopy and how the use of these techniques offers a succinct means of analyzing three-dimensional cellular architecture. We discuss some of the recent work that has taken advantage of these approaches and detail how they have become integral in correlative microscopy workflows.

中文翻译：

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使用X射线断层扫描和相关成像方法测定三维细胞结构。

我们对细胞器和大分子复合物的空间组织以及它们之间的相互作用的大多数理解是使用基于光或电子的微观分析进行成像研究的结果。这些经典的方法虽然很有见地，但仍然受到分辨率限制或生成多维数据的巨大复杂性的限制。X射线的使用和应用在获取微米和纳米断层扫描数据集方面的最新进展提供了一种替代方法，可在纳米级可视化细胞结构。这些新方法可以对未染色的玻璃化细胞进行亚细胞分析，并可以对特定蛋白质靶标进行三维定位，并已成为桥接光电子相关显微镜实验的重要工具。这里，我们回顾了软X射线断层扫描和X射线显微术的理论，仪器详细信息，获取原理和应用，以及这些技术的使用如何为分析三维细胞结构提供了一种简洁的方法。我们讨论了一些利用这些方法的最新工作，并详细介绍了它们如何在相关显微镜工作流程中变得不可或缺。