UNIVERSITY OF TEXAS AT EL PASO

Cryo-Electron Microscopy (cryo-EM) is a technique that can be used for both single particle and tomographic three-dimensional reconstructions of large macromolecular assemblies. Cryo-EM is an essential tool in determining the structure of complexes that fail to crystallize due to their inability to pack into regular arrays or due to their instability and short half-life. Samples are frozen in biologically relevant buffers by flash freezing to liquid nitrogen temperatures, which preserves the biomolecules under conditions that are close to their native environment. Furthermore, this technique spans the gap in the resolution range between light microscopy and X-ray crystallography and Non Magnetic Resonance (NMR). Recent technological advances have produced microscopes with a more coherent electron beam and increased stability that reduces image artifacts. These sophisticated microscopes together with more powerful computers and computer algorithms now routinely allow cryo-EM reconstructions beyond 10 Å resolution for high symmetry macromolecules. Facilities exist for in-house X-ray crystallography, NMR as well as powerful computational resources for structure determination. Furthermore, the University has started construction of a new Chemistry building that includes a cryo-EM facility to house the requested equipment. Construction of the new facility is scheduled to be completed at the same time the proposed instrument will be ready to install. Small molecule and macromolecular X-ray crystallography together with expanded NMR facilities and the new cryo-EM facility will help establish a strong and highly competitive structural biochemistry program at The University of Texas at El Paso (UTEP) and the southwestern region of the U.S. Therefore we would like to propose the acquisition of a FEI Titan Krios electron microscope that will be generally available to researchers. This top of line instrument is equipped with a field emission gun and an accelerating voltage of up to 300 kilovolts. The newly designed sample stage is extremely stable and capable of maintaining the sample at liquid nitrogen temperatures for 24 hours without interruption. The Titan Krios has been designed for maximum stability and has routinely provided atomic resolution data to the materials science field. The addition of cryo capabilities and a larger pole piece has now allowed its use in biological applications. Cryo-EM is a state-of-the-art technique that is not available within 10 hours driving distance of UTEP. The nearest facility equipped with cryo-EM microscopes is 717 miles away in San Diego, California. Boulder and Houston have the next closest facilities and are 733 miles and 756 miles away, respectively. UTEP, a minority serving institution with more than 70% minority enrollment, is committed to establishing a cryo-EM facility to serve the southwestern region that includes West Texas, New Mexico, Arizona, Mexico and South America. A large number of minority students and minority faculty will have access to state-of-the-art technology and facilities where future Structural Biologists can be trained.

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