Towards Correlative Super-Resolution Fluorescence and Electron Cryo-Microscopy
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This peer-reviewed scientific article is freely accessible after registration due to a friendly sponsoring by Leica Microsystems! The Leica EM GP allows plunge freezing of thin-blotted samples and blotting from one side of the specimen holder, and is thus particularly suitable for cellular samples. Additionally a new Leica cryo-stage for upright microscopes was used.
Fast freezing techniques (vitrification) are immobilizing biological samples and are preserving structures in their native state. In this article you find an overview on current developments in super-resolution correlative light and electron microscopy (CLEM), including sample preparation techniques.
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G. Wolff, C. Hagen, K. Grünewald and R. Kaufmann; Towards correlative super-resolution fluorescence and electron cryo-microscopy Biol. Cell (2016) 108, 245–258 DOI: 10.1111/boc.201600008
Correlative light and electron microscopy (CLEM) has become a powerful tool in life sciences. Particularly cryo-CLEM, the combination of fluorescence cryo-microscopy (cryo-FM) permitting for non-invasive specific multi-colour labelling, with electron cryo-microscopy (cryo-EM) providing the undisturbed structural context at a resolution down to the Ångstrom range, has enabled a broad range of new biological applications. Imaging rare structures or events in crowded environments, such as inside a cell, requires specific fluorescence-based information for guiding cryo-EM data acquisition and/or to verify the identity of the structure of interest. Furthermore, cryo-CLEM can provide information about the arrangement of specific proteins in the wider structural context of their native nano-environment. However, a major obstacle of cryo-CLEM currently hindering many biological applications is the large resolution gap between cryo-FM (typically in the range of ∼400 nm) and cryo-EM (single nanometre to the Ångstrom range).
Very recently, first proof of concept experiments demonstrated the feasibility of super-resolution cryo-FM imaging and the correlation with cryo-EM. This opened the door towards super-resolution cryo-CLEM, and thus towards direct correlation of structural details from both imaging modalities.
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