This proposal requests support for the purchase of an Thermo Fisher Glacios electron microscope equipped with a Falcon 3EC direct electron detector and EPU data collection software to be shared by several users at the University of Alabama at Birmingham (UAB). Recent technical developments in cryo-EM have revolutionized structural biology, allowing structures of proteins as small as 100 kDa to be determined to near-atomic resolution, without the need to crystallize. These developments include mechanically and optically stable microscopes with automatic sample changers able to run continuously for several days with minimal supervision, and direct electron detectors (DEDs) with greatly improved resolution performance and low noise (high detective quantum efficiency, DQE) compared to traditional CCD detectors. UAB is currently equipped with an FEI F20 electron microscope with a Gatan K3 direct electron detector. While this instrument is theoretically capable of reaching ?4 resolution, it is incapable of high-throughput operation needed for sample screening and high-resolution data collection. The proposed Glacios instrument will enable high-resolution/high-throughput cryo-EM at UAB. Many investigators at UAB who use structural approaches for their research will be able to take advantage of the new capabilities, leading to accelerated progress on many medically important projects in key areas such as antibacterial drug discovery, emerging and biodefense- related viruses, vaccine development, cystic fibrosis, Parkinson's disease, cancer, and others. The Glacios microscope, the Falcon DED and embedded EPU software constitute a fully integrated system that will be easy to operate and maintain by a variety of users. With the new instrumentation, UAB will be able to attract new investigators who need high-resolution cryo-EM for their research. In The proposed instrumentation will have a transformative impact on research, training and recruitment at UAB.
Electron microscopy has the potential to resolve near-atomic resolution structures of proteins and macromolecular complexes ranging in size from 100 kDa to tens of megadaltons. High- resolution electron microscopy is dependent on highly stable microscopes with automatic sample loaders capable of continuous, unsupervised high-throughput operation, and direct electron detectors with greatly improved performance over traditional CCDs. The Thermo Fisher Glacios instrument we plan to acquire will allow high-resolution/high-throughput structure determination for many projects of medical importance, including infectious disease, emerging pathogens, antibacterial drug and vaccine development, Parkinson's disease, cystic fibrosis, cancer and other systems.
