Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE, UK.
Department of Infectious Diseases, Molecular Virology, Heidelberg University Hospital, 69120 Heidelberg, Germany.
Cell. 2020 Jul 23;182(2):515-530.e17. doi: 10.1016/j.cell.2020.05.051. Epub 2020 Jun 30.
Imaging of biological matter across resolution scales entails the challenge of preserving the direct and unambiguous correlation of subject features from the macroscopic to the microscopic level. Here, we present a correlative imaging platform developed specifically for imaging cells in 3D under cryogenic conditions by using X-rays and visible light. Rapid cryo-preservation of biological specimens is the current gold standard in sample preparation for ultrastructural analysis in X-ray imaging. However, cryogenic fluorescence localization methods are, in their majority, diffraction-limited and fail to deliver matching resolution. We addressed this technological gap by developing an integrated, user-friendly platform for 3D correlative imaging of cells in vitreous ice by using super-resolution structured illumination microscopy in conjunction with soft X-ray tomography. The power of this approach is demonstrated by studying the process of reovirus release from intracellular vesicles during the early stages of infection and identifying intracellular virus-induced structures.
跨分辨率尺度的生物物质成像是一项具有挑战性的工作，需要确保从宏观到微观层面的主体特征具有直接且明确的相关性。在这里，我们展示了一种专门用于在低温条件下通过 X 射线和可见光对细胞进行三维成像的相关成像平台。快速冷冻保存是目前用于 X 射线成像中超微结构分析的生物样本制备的黄金标准。然而，大多数低温荧光定位方法都受到衍射限制，无法提供匹配的分辨率。我们通过开发一种集成的、用户友好的平台来解决这一技术差距，该平台用于通过超分辨率结构光照明显微镜结合软 X 射线断层扫描对玻璃态冰中的细胞进行三维相关成像。通过研究感染早期细胞内囊泡中呼肠孤病毒的释放过程并鉴定细胞内病毒诱导的结构，证明了这种方法的强大功能。
