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

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.