Imaging techniques available in the facility
The CryoEM facility provides tailored support for a wide variety of scientific questions requiring high-resolution imaging. Techniques that are currently available at the facility are:
- Standard 2D projecting imaging of thin (cryo-) samples provides structural information with a high resolution.
- Single-particle analysis. Provides ultra-high resolution structure of a single protein or protein complex. Single-particle analysis is the core strength of the facility. First, the quality of the sample is assess with negative stain imaging and 2D class averages are obtained, after which the sample is vitrified and screened until the right conditions are reached to collect data. The data is then processed to produce a 3D structure of the particle. When necessary, samples can be taken to national facilities, such as the SCMI or eBIC.
- (Cryo-) Electron tomography. Provides 3D information of a single specimen by physically tilting the specimen inside the EM. Can be used on cryo-samples as well as plastic sections of cellular material.
- Electron Diffraction. Provides structural information on crystalline material
The facility is working to offer several correlative workflows to combine fluorescence microscopy with EM.
Thermo Fisher Scientific Tecnai F20 electron microscope (200 kV, field emission gun) equipped with an 8k x 8k CMOS camera (TVIPS F816).
Gatan single tilt liquid nitrogen cryo-transfer holder (Gatan 626), with transfer-station, pumping station and temperature control unit.
Thermo Fisher Scientific Vitrobot automated vitrification unit (Vitrobot mark IV).
Various supporting equipment, such as a glow-discharge unit, negative stain facilities and a room-temperature holder.
Extensive computer resources for data processing.
The electron microscope is located in the Daniel Rutherford Building, room G.01, Max Born Crescent, The King's Buildings, EH9 3BF Edinburgh.
Booking is done via the system PPMS (Stratocore): Here
The CryoEM facility provides extensive training for users, ranging from negative stain sample preparation, conventional room-temperature TEM, to plunge-freezing, cryoTEM to data-processing of single-particle analysis.
To ensure that the cryoEM Facility can operate and keep operating successfully, we ask that the facility is acknowledged in any publications, posters and talks that uses data acquired in the facility. Even though the facility offers a paid service, it is still dependent on financial support from the Wellcome Centre and the School. Furthermore, for larger equipment upgrades the facility is dependent on grants. For all of these, the facility has to show its scientific output.
When data is acquired in the facility, samples are screened in the facility prior to high-resolution data collection and/or samples are prepared in the facility, please mention the facility in the acknowledgments:
Data/Grid screening was acquired/performed in the cryoEM facility in School of Biological Sciences at the University of Edinburgh. The cryoEM facility was set up with funding from the Wellcome Trust (087658/Z/08/Z) and SULSA.
Please feel free to adjust the statement to fit the situation in the publication.
When data and/or images are used that have been acquired by a staff-member of the facility, or data from samples that have been prepared by a staff-member, please consider offering a co-authorship to the member of staff.
Similarly, when a member of staff has been closely involved in the project, and made substantial scientific input, please consider offering a co-authorship.
For the acknowledgement:
Maarten W. Tuijtel (MWT) is supported by School of biological sciences at the University of Edinburgh and Wellcome Centre for Cell Biology