The EM facility provides equipment and expertise in the field of cryo-electron microscopy, particularly single-particle analysis for high-resolution structure determination, but also cellular imaging by TEM (Transmission EM) and SEM (Scanning EM).
All the stages in project development preceding high-resolution data collection can be performed at the EM facility. With our Talos F200i TEM and a Falcon 3EC direct electron detector, the users can analyse negatively-stained samples and collect cryo-EM data to solve sub-nanometric structures.
We also have equipment for ultrathin sectioning of resin-embedded cellular specimens. They can be prepared and imaged using our Apreo Volumescope SEM. Users can also take part in the development of the correlated light and electron microscopy (CLEM) capabilities with the Leica Thunder Imager.
The facility provides training, advice and help for the data collection and image processing workflow for 3D structure determination. The facility has a dedicated GPU workstation for newcomers in data processing and has access to the High-Performance Computer cluster JEX.
Transmission electron microscopy:
Scanning electron microscopy:
Saleh A, Noguchi Y, Aramayo R, Ivanova ME, Stevens KM, Montoya A, Sunidhi S, Carranza NL, Skwark MJ, Speck C. (2022) The structural basis of Cdc7-Dbf4 kinase dependent targeting and phosphorylation of the MCM2-7 double hexamer. Nat Comm 13(1), 2915.
Gutierrez-Escribano P, Hormeño S, Madariaga-Marcos J, Solé-Soler R, O’Reilly FJ, Morris K, Aicart-Ramos C, Aramayo R, Montoya A, Kramer H, Rappsilber J, Torres-Rosell J, Moreno-Herrero F, Aragon L. (2020). Purified Smc5/6 Complex Exhibits DNA Substrate Recognition and Compaction Affiliations. Mol Cell 80(6), 1039-1054.
Yuan Z, Schneider S, Dodd T, Riera A, Bai L, Yan C, Magdalou I, Ivanov I, Stillman B, Li H, Speck C. (2020). Structural mechanism of helicase loading onto replication origin DNA by ORC-Cdc6. Proc Natl Acad Sci USA 117(30), 17747-56.
Gutierrez-Escribano P, Newton MD, Llauró A, Huber J, Tanasie L, Davy J, Aly I, Aramayo R, Montoya A, Kramer H, Stigler J, Rueda DS, Aragon L. (2019). A conserved ATP- and Scc2/4-dependent activity for Cohesin in tethering DNA molecules. Sci Adv 5, 6804.
Noguchi Y, Yuan Z, Bai L, Schneider S, Zhao G, Stillman B, Speck C, Li H. (2017). Cryo-EM structure of Mcm2-7 double hexamer on DNA suggests a lagging strand DNA extrusion model. Proc Natl Acad Sci USA 114, 9529-38.