细胞冷冻光电关联技术的实施与探讨

Implementation and Discussion of Cryo-Correlative Light and Electron Microscopy of Cells

  • 摘要: 光电关联技术(CLEM)是一项将光学显微镜和电子显微镜相结合的新兴技术,该技术兼具了光学成像的功能性优势和电子显微成像的高分辨优势. 冷冻光电关联技术(Cryo-CLEM)是在冷冻条件下实施的光电联用技术,其能在接近天然状态下成像,并能利用冷冻电子断层成像技术(Cryo-ET)获取细胞原位高分辨结构. 为了确定Cryo-CLEM技术实施的关键步骤及注意事项,对细胞培养、冷冻样本制备、冷冻光学成像、样本减薄及关联策略等进行探索,初步解决了Cryo-CLEM技术实施的关键和难点,旨在为相关领域的研究提供应用范例.

     

    Abstract: Correlative light and electron microscopy (CLEM) is an emerging technology that combines the functional advantages of light microscopy with the high-resolution advantages of electron microscopy. Cryo-correlative light and electron microscopy (Cryo-CLEM) is a kind of CLEM under freezing conditions, which can take imaging near the natural state and obtain the in-situ high-resolution structure of cells by cryo-electron tomography (Cryo-ET). In order to implement the Cryo-CLEM technology, the key steps and considerations, such as cell culture, cryo-sample preparation, cryo-optical imaging, sample thinning, and correlation strategies were explored. The key and difficult points in the implementation of Cryo-CLEM technology were preliminarily addressed, aiming to provide an application examples for research in related fields.

     

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