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Institut de Biologie StructuraleGrenoble / France

Contact person(s) related to this article / NEUMANN Emmanuelle
Contact person(s) related to this article / Benoit Gallet

Microscope Tecnai 12 (FEI)

Tecnai12 120 kV

The Tecnai12 Microscope operating at 120 kV is dedicated to negative-staining and quality control.
Tomography at Room Temperature is available.

Specificities :
- Tension : 80 - 120 kV
- Electron source : LaB6
- Orius 600 & 1000 GATAN cameras
- Holders : single tilt, FISCHIONE 2020 tomography holder, rotation holder

Microscope Tecnai F20 (FEI)

Tecnai F20 200 kV

The Tecnai F20 transmission electron microscope operating at 200 kV (FEG) is dedicated to cryo-electron microscopy.

Specificities :
- Tension : 80 - 200 kV
- Field Emission Gun (FEG)
- Windows 7
- 4kx4k Ceta FEI camera
- Holders : single tilt, FISCHIONE 2020 tomography holder, 626 GATAN cryo-holder, 914 cryo-tomo holder.

Microscope Tecnai POLARA (FEI)

Polara FEG Microscope - 300 kV

Specificities :
- Tension : 100 - 300 kV
- Field Emission Gun (FEG)
- K2 Summit GATAN electron counting direct detection camera
- 2kx2k GATAN GIF
- Software : Tomography FEI (FEI), Xplore3D (FEI), GMS 3.2 (GATAN - automatic data collection)

Since February 2010, the IBS has a Polara transmission electron microscope for high-resolution cryo-electron microscopy. The microscope was funded by generous grants from IBISA, CNRS, EMBL and the Région
Rhône-Alpes and its acquisition has only been possible because IBS (UMR 5075), UVHCI (UMI 3265) and EMBL Grenoble Outstation have put effort together to get it. Electron microscopists from the Structural Biology Partnership have access to this microscope.
This microscope is a cryo-transmission EM with a 300 kV field emission gun (FEG), which allows imaging of proteins in their native state. The microscope stage is stable to allow high-resolution imaging. The design «cryo» also gives a low rate of specimen contamination in the microscope (one can work on the same grid for several days).
The Polara can be loaded with 6 samples, for convenient screening of multiple samples. The microscope has reliable grid loading and specimen transfer (achieved through a protected environment in the loading bay, and the transfer system under vacuum).

To collect high resolution data, the microscope is now fitted with one of the highest performance detector available for cryo-electron microscopy: the K2 Summit electron counting direct detection camera (GATAN). The Tecnai G2 Polara is also fitted with a 2k x 2k Gatan post column energy filter.

The Polara is also equipped to do single tilt electron tomography (collecting a series of tilted images +/-70 deg) with:

- The following tomography imaging options:
Gatan K2 Summit,
Gatan Imaging Filter 2002 (2k X 2k Tridiem GIF)

- The following software:
FEI Tomography (FEI),
Xplore3D (FEI)
GMS 3.2 (automatic data collection - GATAN)

The Polara microscope is open to outside (national and international) users after approval of their scientific projects. Trained cryo-electron microscopists have priority but other projects can be accommodated within the framework of collaboration. In any case if you want to use or have an access to the Polara Microscope or to another equipment of the electron microscopy platform, please send an email to : ibs-plateforme-em.contact@ibs.fr.

Microscope associated instruments

Carbon evaporator

Instrument: Emitech K950X
Application: Deposits an homogeneous carbon film onto the surface of a mica sheet or on formvar grids.

Glow discharger

Instrument: Emitech K100X
Application: Makes the surface of the grid hydrophilic/hydrophobic by charging negatively/positively the carbon-coated grid.

Freezing automated system

Instrument: FEI Vitrobot MARK IV
Application: Vitrification of sample by plunge-freezing (preparation of cryo-EM grids).

Film scanner

Instrument: Zeiss/Intergraph Photoscan TD
Application: High resolution scanning of electron micrographs.

Instruments for cellular sample preparation

Automated high-pressure freezer

Instrument: Leica EM-HPM100
Application: Cryo-fixation of biological sample under high pressure (2000 Bars).

Automated freeze substitution machine

Instrument: Leica EM-AFS2 + EM-FSP
Application: Freeze substitution process or Progressive Lowering of Temperature (PLT) for ambient or low-temperature embedding of sample, manually or automated with the FSP robot.

(Cryo-)ultra-microtome

Instrument: Leica EM-UC7 + EM-FC7
Application: Ultra-thin sectionning of embedded biological samples, at room temperature or in cryo-conditions.

Automated immuno-labeling machine

Instrument: Leica EM-IGL
Application: Automated staining or immuno-labelling steps on samples on grids (single particles or sections).