Grid and Sample Preparation for Cryo-EM
The BSC can assist in the preparation of grids for the collection of cryo-EM single particle analysis (SPA) data. The facility has a Pelco easiGlow discharge system for grid preparation and a Leica EM-GP2 cryo plunger as well as a ThermoFisher Vitrobot for freezing samples. An assortment of commonly used cryo-EM grids is available in the BSC. The BSC can assist with shipping of samples to cryo-EM data collection facilities, as well as data transfer and processing. Individual researchers will need to apply for data collection time for their projects at a national cryo-EM center.
The cryoplungers and glow discharger are located in room 2750 of the Stephenson Life Sciences Research Center on the University of Oklahoma-Norman Research Campus.
Scheduling for the use of the grid preparation equipment can be done by contacting the facility director, Dr. Len Thomas, lmthomas@ou.edu, (405) 325-1126.
Cryo-EM Sample Screening/Data Collection on Tundra
In collaboration with OU’s Samuel Roberts Noble Microscopy Laboratory (SRNML), the BSC can assist in screening of cryo-EM grids and data collection on OU’s ThermoFisher FEI Tundra 100 kV Single Particle Cryo-TEM.
The Tundra is located in room 1655 of the Stephenson Life Sciences Research Center on the University of Oklahoma-Norman Research Campus.
Please contact the BSC facility director, Dr. Len Thomas, lmthomas@ou.edu, (405) 325-1126 or the SRNML TEM specialist, Dr. Julian Sabisch, jsabisch@ou.edu for more information.
Applicable fees will be charged for equipment training, use, and services: grid/sample preparation fees and Tundra usage fees.
Funding for the instrumentation was provided, in part, by an Institutional Development Award (IDeA) from the NIH (Award P20GM103640 and P30GM145423, PI: Dr. Ann West).
GPU Workstations for Cryo-EM Data Processing
Three GPU workstations are available for cryo-EM data processing (80+ TB each). Workstations have a full suite of programs for cryo-EM processing, structure determination and refinement (CryoSPARC, RELION, and PHENIX).
Data may be stored for 90 days. Once data are collected, data should be transferred to researchers’ systems/choice of data storage. Research groups are responsible for the long-term archiving of their data (OSCER is available). Please talk to the facility director about data storage, if needed.
Please contact the core facility director, Dr. Len Thomas, (405) 325-1126, for access to the workstations.
How to acknowledge grid preparation and/or cryoEM data collection using the Tundra:
Cryo-EM grid preparation was carried out at the Biomolecular Structure Core, University of Oklahoma–Norman, with support from an Institutional Development Award from the National Institutes of Health (P30GM145423). Microscopy data collection using the Tundra TEM was supported by the Samuel Roberts Noble Microscopy Laboratory, the OU Office of the Vice President for Research and Partnerships, and NIH award P30GM145423.
