The cryo-electron microscopy (cryo-EM) facility, established in 2016, counts with state-of-the-art equipment for preparation and imaging of vitrified biological macromolecule specimens. The goal of the facility is to provide access to cutting-edge technology and know-how for determination of high-resolution three-dimensional macromolecular structures.
The centerpiece of the cryo-EM facility is a 200 kV ThermoFisher Talos Arctica X-FEG transmission electron microscope (TEM) outfitted with an Autoloader cryo-specimen handling system, a CETA CMOS camera and a Gatan K3 Summit direct electron detector (DED). A recently implemented image shift data collection protocol (including stigmation and coma corrections) enables acquisition of up to 4000-5000 DED movies every 24 hrs. The facility also counts with a 120kV ThermoFisher Talos L120C TEM for low-resolution imaging of stained specimens and cryo-specimen screening. A Gatan Solarus 950 plasma cleaning system, an FEI Vitrobot Mark IV, a manual station for grid vitrification, glow discharge and carbon evaporation systems, and all other ancillary equipment required for cutting-edge cryo-EM work are also available.
Cryo-EM data collection is fully automated using the Leginon system and all image data is directed to a web-accessible database that makes possible remote monitoring of data collection and on-the-fly preliminary image analysis (frame alignment, image evaluation, particle picking, etc.) through Appion and CryoSPARC Live. We have recently enabled remote data collection, which allows users to monitor microscope status, select targets for imaging and control data collection through an internet connection.
The facility is located in the heart of the Anschutz Medical School Campus, on the ground level of the RC-1 South Building (Rm 1301B). It is directed by Francisco Asturias and managed by Eduardo Romero Camacho, both in the Department of Biochemistry & Molecular Genetics.
This facility is supported in part by the University of Colorado School of Medicine and the University of Colorado Cancer Center.
All publications resulting from the use of the Cryo-EM Core Facility should acknowledge the Cancer Center Support Grant (P30CA046934) and submit the publication to PubMed Central.
Facility Director
Office: Research 1 South L18-10104
francisco.asturias@cuanschutz.edu
303-724-1624
Facility Manager
Office: Research 1 South L18-10401D
russell.mcfarland@cuanschutz.edu
206-605-7028
University of Colorado School of Medicine
Research 1 South
L18-1301B
12801 E. 17th Ave.
Aurora, CO 80045