Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA 94158, United States; National Center for X-ray Tomography, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States.
Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA 94158, United States; Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States; National Center for X-ray Tomography, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States.
Curr Opin Struct Biol. 2019 Oct;58:324-332. doi: 10.1016/j.sbi.2019.06.012. Epub 2019 Sep 6.
Cellular complexity is represented best in high-spatial resolution, three-dimensional (3D) reconstructions. Soft X-ray tomography (SXT) generates detailed volumetric reconstructions of cells preserved in a near-to-native, frozen-hydrated state. SXT is broadly applicable and can image specimens ranging from bacteria to large mammalian cells. As a reference, we summarize light and electron microscopic methods. We then present an overview of SXT and discuss its role in cellular imaging. We detail the methods used to image biological specimens and present recent highlights that illustrate the capabilities of the technique. We conclude by discussing correlative imaging, specifically the combination of SXT and fluorescence microscopy performed on the same specimen. This correlated approach combines the structural morphology of a cell with its physiological characteristics to build a deeply informative composite view.
细胞复杂性在高空间分辨率的三维（3D）重建中表现得最好。软 X 射线断层扫描（SXT）可以对保存在近乎自然的冷冻水合状态下的细胞进行详细的体积重建。SXT 应用广泛，可以对从细菌到大型哺乳动物细胞的标本进行成像。作为参考，我们总结了光学和电子显微镜方法。然后，我们介绍了 SXT 的概述，并讨论了它在细胞成像中的作用。我们详细介绍了用于对生物标本进行成像的方法，并介绍了一些最近的亮点，这些亮点说明了该技术的能力。最后，我们讨论了相关成像，特别是在同一标本上同时进行 SXT 和荧光显微镜的组合。这种相关的方法将细胞的结构形态与其生理特征结合起来，构建了一个信息量丰富的综合视图。
