We are requesting funds to purchase a Gatan K2 Summit direct electron detector that will support the research programs of nine NIH-funded investigators at the University of Texas Medical School at Houston. This detector will replace an outdated and unserviceable Tietz 4k x 4k CCD presently interfaced with a Polara G2 300 kV FEG cryoelectron microscope. This state-of-the-art K2 Summit detector has significantly improved signal-to-noise, fast read-out rates and has electron-counting capabilities that will provide investigators unprecedented resolution and throughput required for supporting high-resolution single particle, 2D electron crystallography and electron cryo-tomography projects. Data from the K2 detector will be collected on a separate high-end workstation processed through the Summit's high-speed data chassis capable of read-out rates of up to 400 frames per second. The detector will be integrated into the existing Polara G2 electron microscope and will be managed using the well-established infrastructure already in place. There is no other direct detector (or cryo-electron microscope) in the UT Health Science Center capable of supporting these projects. The total projected usage from the four major investigators is ~80%, leaving ~20% time for the five minor investigators and for supporting other new potential projects requiring cryoelectron microscopy. The Institution has made a major financial commitment of $40,000 per year for five years to support the projected maintenance costs of the new direct detector. In total, we have a cohort of well-funded NIH investigators relying on an unserviceable and out-of-date 9-year-old CCD camera. All of their research programs employ the unique advantages of cryo-EM to enable correlations between molecular mechanism and function for a host of basic prokaryotic and eukaryotic cell biological and biochemical processes that carry significant health- related impacts. The requested Gatan K2 Summit direct detector will enormously facilitate the pace of their research programs and provide structural insights not previously achievable.
|Fischer, Tara D; Hylin, Michael J; Zhao, Jing et al. (2016) Altered Mitochondrial Dynamics and TBI Pathophysiology. Front Syst Neurosci 10:29|
|Morado, Dustin R; Hu, Bo; Liu, Jun (2016) Using Tomoauto: A Protocol for High-throughput Automated Cryo-electron Tomography. J Vis Exp :e53608|
|Farley, M M; Swulius, M T; Waxham, M N (2015) Electron tomographic structure and protein composition of isolated rat cerebellar, hippocampal and cortical postsynaptic densities. Neuroscience 304:286-301|
|Hu, Bo; Morado, Dustin R; Margolin, William et al. (2015) Visualization of the type III secretion sorting platform of Shigella flexneri. Proc Natl Acad Sci U S A 112:1047-52|
|Fan, Guizhen; Baker, Matthew L; Wang, Zhao et al. (2015) Gating machinery of InsP3R channels revealed by electron cryomicroscopy. Nature 527:336-41|
|Hu, Bo; Margolin, William; Molineux, Ian J et al. (2015) Structural remodeling of bacteriophage T4 and host membranes during infection initiation. Proc Natl Acad Sci U S A 112:E4919-28|
|Baker, Mariah R; Fan, Guizhen; Serysheva, Irina I (2015) Single-Particle Cryo-EM of the Ryanodine Receptor Channel in an Aqueous Environment. Eur J Transl Myol 25:4803|