Super-resolution Cryo-Correlative Light and Electron Microscopy to studying macromolecular structure in the cell nanocontext

为了了解复杂的生物结构和功能，我们需要更全面的成像策略。低温相关光电子显微镜（cryo-CLEM）已成为生物成像方法和发展的前沿。冷冻电子显微镜（cryo-EM）是获得亚纳米分辨率分子和细胞超微结构的有力工具。冷冻荧光显微镜（cryo-FM）在研究细胞中特定生物分子的分布和组织方面发挥着主导作用。Cryo-CLEM是一种将复杂生物系统中的功能与结构联系起来的强大方法;然而，一个主要缺点是cryo-EM（纳米尺度）和cryo-FM（几百个数字尺度）之间的分辨率差异。我们建议开发超分辨率cryo-FM，从而发展一种新的成像方法，对保存在近天然状态的生物样品提供大分子水平的光学分辨率。这一发展将为部署高要求的超分辨率cryo-CLEM铺平道路，使各种相关的生物结构和功能研究成为可能。

To understand complex biological structure and function, we need more comprehensive imaging strategies. Cryo-correlative light and electron microscopy (cryo-CLEM) has been put into the frontline of bio-imaging method and development. Cryo-electron microscopy (cryo-EM) is a powerful tool to obtain up to sub-nanometre resolution molecular and cellular ultrastructure. Cryo-fluorescence microscopy (cryo-FM) takes the leading role to study the distribution and organisation of specific biomolecules in cells. Cryo-CLEM is a powerful way to link function to structure in complex biological systems; however a major drawback is the discrepancy in resolving power between cryo-EM (nanometre scale) and cryo-FM (a few hundred number scale). We propose to develop super-resolution cryo-FM, leading to the development of a new imaging method offering macromolecular level optical resolution on bio-samples preserved in a near-native state. The development will pave the way for the deployment of highly demanded super-resolution cryo-CLEM, enabling a variety of correlative biological structure and function studies.