Schaller Research Groups, Department of Infectious Diseases, Virology, Heidelberg University Hospital, 69120 Heidelberg, Germany.
Schaller Research Groups, Department of Infectious Diseases, Virology, Heidelberg University Hospital, 69120 Heidelberg, Germany.
Curr Opin Virol. 2023 Aug;61:101338. doi: 10.1016/j.coviro.2023.101338. Epub 2023 Jun 20.
Cellular cryo-electron tomography (cryo-ET) offers 3D snapshots at molecular resolution capturing pivotal steps during viral infection. However, tomogram quality depends on the vitrification level of the sample and its thickness. In addition, mandatory inactivation protocols to assure biosafety when handling highly pathogenic viruses during cryo-ET can compromise sample preservation. Here, we focus on different strategies applied in cryo-ET and discuss their advantages and limitations with reference to severe acute respiratory syndrome coronavirus 2 studies. We highlight the importance of virus-like particle (VLP) and replicon systems to study virus assembly and replication in a cellular context without inactivation protocols. We discuss the application of chemical fixation and different irradiation methods in cryo-ET sample preparation and acquisition workflows.
细胞冷冻电子断层扫描（cryo-ET）以分子分辨率提供 3D 快照，可捕获病毒感染过程中的关键步骤。然而，断层图像的质量取决于样品的玻璃化水平及其厚度。此外，在冷冻电子断层扫描过程中处理高致病性病毒时，为确保生物安全而采取的强制性灭活方案可能会损害样品的保存。在这里，我们重点介绍了冷冻电子断层扫描中应用的不同策略，并参考严重急性呼吸系统综合征冠状病毒 2 研究讨论了它们的优点和局限性。我们强调了病毒样颗粒（VLP）和复制子系统的重要性，这些系统可在不使用灭活方案的情况下，在细胞环境中研究病毒的组装和复制。我们讨论了化学固定和不同辐照方法在冷冻电子断层扫描样品制备和采集工作流程中的应用。
