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FRENCH INFRASTRUCTURE FOR INTEGRATED STRUCTURAL BIOLOGY

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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 CMOS (FEI FALCON3 4K*4K and GATAN K3) high-sensitivity direct electron detector cameras, Cs corrector, phase plate and GIF.

The Glacios cryo-electron microscope allows high-resolution SPA and CET automated data collection. Its FEG is usually tuned to 200keV. An automatic loading mechanism allows mounting twelve grids at a time. This microscope is equipped with two digital cameras: FEI Ceta-D and the direct electron detector GATAN K2

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.

Grid preparation:

1. Chameleon: NEW EQUIPMENT

Sample vitrification is a critical step in the cryo-EM workflow, and the bottleneck for many projects. While the classical plunge freezing method is still widely used, it suffers from major limitations such as preferential orientation and protein denaturation which can hinder the success of many projects. The Chameleon offers a new, fast and blot-less freezing approach to tackle challenging samples for structure determination by cryo-EM. This fully automatized vitrification device limits the manual handling of the grids for improved consistency in grid preparation. It also offers direct visualization of the grids for easier control of grid quality.

2. Vitrobot:

For purified samples (not yet on cryo-EM grids) we offer freezing (Vitrobot system) and screening of grids. 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).

The non-infectious character of the samples needs to be confirmed by the director of the applying institute.

Please include a short description of the project and the characterisation of the sample (biophysically characterisation and images / class averages of cryo-EM tests if available). Applications will be handled confidentially as for any applications, submit proposal and select the platform  "cryo electron microscopy (Instruct-centre France 1 IGBMC)"


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

See more at https://instruct-eric.eu/platform/electron-microscopy-strasbourg-france/


3rd Instruct-ERIC Best Practices in CryoEM Workshop organised in November 2019

https://instruct-eric.eu/content/3rd-instruct-best-practices-in-cryoem-workshop


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