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FRENCH INFRASTRUCTURE FOR INTEGRATED STRUCTURAL BIOLOGY

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INSTRUCT CENTER - FRANCE 1 - IGBMC

CRYO ELECTRON MICROSCOPY

The Titan Krios cryo electron microscope is the latest generation electron microscope with capacities for high-resolution data collection for both Cryo Electron Tomography (CET) experiments and Single Particle Analysis (SPA) with automated data collection. The electron source is a Field Emission Gun (FEG) that can be operated at 80keV, 100keV, 200keV or 300keV. An automatic loading mechanism allows mounting twelve grids at a time. This microscope is equipped with a CMOS (FEI FALCON 4K*4K) high-sensitivity direct electron detector camera.

The Tecnai F30 Polara cryo electron microscope allows high-resolution SPA and CET automated data collection. Its FEG is usually tuned to 100keV, 200keV or 300keV. The grids are manually mounted 6 at a time. This microscope is equipped with 3 digital cameras: FEI CMOS 4K*4K "FALCON", FEI CCD 4K*4K "EAGLE" and a GATAN CCD 2K*2K "ORIUS".

The Tecnai F20 is equipped with a FEG operated at 100keV or 200keV. Grids are mounted one at a time using a Gatan side-entry cryo-holder. This microscope is equipped with one digital camera (GATAN CCD 2K*2K "US10001"), it is used to collect data for cryo-SPA or room temperature electron tomography using a Fischion side-entry holder.

Cryo-EM grids are flash-frozen in a temperature and humidity controlled environment using a Vitrobot system (FEI).

Although all our microscopes are equipped with digital cameras, we can still record images on films and scan them with a high-resolution Heidelberger Druckmaschinen drum scanner ( 5 micron pixelsize).

NOTE: when asking access for Titan or Polara, please add to your proposal preliminary image of the sample in cryo condition.

Availability of the microscopes at: http://em-reservation.igbmc.fr.

See more at https://www.structuralbiology.eu/update/platform/cryo-electron-microscopy--on-polara-f30-at-igbmc-cerbm-strasbourg/

https://www.structuralbiology.eu/update/platform/cryo-electron-microscopy--on-titan-krios-at-igbmc-cerbm-strasbourg

 

 

BIBLIOGRAPHY

Structure of the human 80S ribosome. H. Khatter, A. G. Myasnikov, K. Natchiar & B. P. Klaholz. Nature 2015, in press.

Cryo-Electron Microscopy Structure of Lactococcal Siphophage 1358 Virion. Spinelli S, Bebeacua C, Orlov I, Tremblay D, Klaholz BP, Moineau S, Cambillau C. J Virol. 2014 Aug;88(16):8900-10. doi: 10.1128/JVI.01040-14. Epub 2014 May 28.

Formation of circular polyribosomes on eukaryotic mRNA without cap-structure and poly(A)-tail: a cryo electron tomography study. Afonina ZA1, Myasnikov AG2, Shirokov VA1, Klaholz BP2, Spirin AS3. Nucleic Acids Res. 2014 Aug;42(14):9461-9. doi: 10.1093/nar/gku599. Epub 2014 Jul 12.