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The outbreak of a novel coronavirus (2019-nCoV) represents a pandemic threat that has been declared a public health emergency of international concern. The CoV spike (S) glycoprotein is a key target for vaccines, therapeutic antibodies, and diagnostics. To facilitate medical countermeasure development, we determined a 3.5-angstrom-resolution cryo-electron microscopy structure of the 2019-nCoV S trimer in the prefusion conformation. The predominant state of the trimer has one of the three receptor-binding domains (RBDs) rotated up in a receptor-accessible conformation. We also provide biophysical and structural evidence that the 2019-nCoV S protein binds angiotensin-converting enzyme 2 (ACE2) with higher affinity than does severe acute respiratory syndrome (SARS)-CoV S. Additionally, we tested several published SARS-CoV RBD-specific monoclonal antibodies and found that they do not have appreciable binding to 2019-nCoV S, suggesting that antibody cross-reactivity may be limited between the two RBDs. The structure of 2019-nCoV S should enable the rapid development and evaluation of medical countermeasures to address the ongoing public health crisis.
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Funding to establish the cryo-EM core facility was provided by the National Science Foundation, the Murdock Charitable Trust, and MSU's Office of Research, Economic Development, and Graduate Education
Additionally, the Molecular Electron Microscopy Suite (MEMS) at Harvard Medical School is a separate user resource currently available to qualified researchers. This user facility offers training and supervision in negative-stain and cryo-transmission electron microscopy. Equipment includes three transmission electron microscopes, two cryo plungers, and sample preparation areas.
Cryo-electron microscopy (cryo-EM) is rapidly becoming the method of choice for biochemistry labs around the world, helping accelerate research. With cryo-EM, molecular details can be seen at biologically relevant resolutions, providing insights into protein function and disease mechanisms, and facilitating effective drug design. Recent advances in instrument design and automation mean cryo-EM is easier to use and more cost effective than ever before. With cryo-EM, you can gain deeper insights into complex biological processes and pathways. While researchers use a variety of techniques to determine protein structure, cryo-EM can provide details on protein function that other methods simply cannot.
Single particle analysis is a cryo-electron microscopy technique that enables structural characterization at near-atomic resolutions, unraveling dynamic biological processes and the structure of biomolecular complexes/assemblies.
Volume electron microscopy (vEM) is a group of techniques that reveals the 3D ultrastructure of cells and tissues at micron to millimeter volume scales, at nanometer-level resolutions, and even at native state under cryogenic conditions.
Successful protein preparation lies at the base of any successful structural biology technique. For cryo-electron microscopy, a wide range of solutions is required to achieve the highest sample quality before freezing of the specimen on the EM grid.
To understand protein function, you need complex and structure information beyond individual proteins. Integrative structural biology combines mass spectrometry and cryo-EM for the determination of large dynamic complex structures.
The David Van Andel Advanced Cryo-Electron Microscopy Suite is a state-of-the-art facility that offers cost-effective cryo-EM and biological EM services along with consulting and hands-on training to internal and external investigators.
Linux workstations are an indispensable part of the cryo-EM image processing pipeline. The Core has three Linux workstations (Silicon Mechanics Workform 2000.v6) dedicated to cryo-EM analysis that are reserved for core users.
To meet the greater demand for cryo- and biological electron microscopy, we provide a wide range of cost-effective and efficient EM services to VAI investigators, external academic institutes and commercial labs.
To put its impact in context, in 2007 fewer than 10 molecular structures at less than 5 angstroms (a very tiny unit of measurement) were reported in the literature. Thanks to improvements in cryo-EM technology, by 2015 that number had shot up to nearly 200 (Egelman EH et al; 2016).
Titan Krios is a state-of-the-art sophisticated high-voltage (300kV) transmission electron microscope. It has a superb electronic optical system and a capability to do fast high-throughput data collection. Titan Krios is equipped with the latest Gatan Image Filter (GIF) and Gatan K2 Summit direct electron detector. This combination allows better electron signal to be recorded. This configuration is currently the ideal setup for high-resolution structure visualization, using single-particle electron cryomicroscopy and electron tomography.
Both microscopes are equipped with a Volta Phase Plate to enable visualization of smaller macromolecular complexes. The facility has additional equipment for sample preparation and storage, including two FEI Vitrobot Mark IV, a glow discharge unit, an auto-filled 55-cylinder cryo-grid storage system etc.
The program is broadening access to high-resolution cryoelectron microscopy (cryoEM) and tomography (cryoET) for biomedical researchers by creating national service centers, and cultivating a skilled workforce through the development and implementation of cryoEM training material. CryoEM and cryoET enable high-resolution, three-dimensional data collection on samples that could not be used with other techniques in the past, such as samples containing mixtures of different biological structures or intact cells. The three National cryoEM Service Centers offer usage of state-of-the-art equipment, technical support, and cross-training for the production and analysis of high-resolution data. These offerings are available at no charge for non-profit use, eliminating the high cost barrier usually associated with cryoEM. The NIH recently funded four centers to make up the National Network for CryoET. This Network will provide the biomedical research community access to advanced instrumentation for cryoET, cryoET specimen preparation, and collection of high-resolution cryoET data as well as cross-training in cryoET methods. Check back for updates on this exciting new resource!
Open access instructional material on cryoEM for those with or without a structural biology background is already available through the CryoEM Curriculum Development sites. Further developments are expected soon.
Did you miss one of the free monthly webinars on cryoEM current practices and strategies hosted by the National Centers for Cryoelectron Microscopy? Catch up before the next one by watching recordings!
CEMAS is at the forefront in the convergence of physical and life sciences. Our multidisciplinary approach drives synergy and amplifies characterization capabilities to challenge "what is possible" in electron microscopy. Our comprehensive cryo-electron microscopy (cryo-EM) program advances this effort.
Located in a custom-designed facility, all cryo-EM instruments at CEMAS meet or exceed manufacturer performance specifications. CEMAS recently installed a Thermo Scientific Glacios Cryo-TEM with a Falcon III direct electron detector and a Ceta-D camera for MicroED analysis. In addition, CEMAS has a Thermo Scientific Krios G3i Cryo-TEM equipped with a Bioquantum imaging filter and a K3 direct electron detector, imaging phase plates, STEM/TEM tomography, and an image spherical aberration corrector.
CEMAS can assist users in all aspects of the cryo-EM workflow, including initial negative stain analysis, cryogenic sample preparation, grid screening, high-resolution data collection, and preliminary data analysis.
Researchers at Ohio State have organized an informal Cryo-EM Interest Group that meets 1-2 times per month. Meetings consist of a highlight talk lasting approximately 30 minutes, followed by a discussion about data analysis, sample preparation, and other cryo-EM focused topics. The group also welcomes outside speakers to talk about their research.
Reaching out to the community because I have been having streaky artifacts with a lot of separate datasets recently that have me kind of stumped. For a little bit of background: I have this issue with data collected on two different Krioses (each with a K3 camera). I work nearly exclusively with ribosomes on UltrAuFoil grids that I float my own amorphous carbon onto (one exception where the holey film was carbon instead of gold). My typical workflow jumps around between relion and cryosparc and I see issues regardless of software, but cryosparc provides a bit more visual feedback that I thought may be helpful to share here.
The Cryo-EM (cryogenic electron microscopy) Core Facility at UTHealth is a state-of-the-art resource in the Texas Medical Center in Houston, Texas. The facility was established to meet the needs of researchers in Houston and nationwide who do not have adequate access to the latest cryo-EM instrumentation and expertise. Recent advances in cryo-EM technology have opened a new door in structural biology attracting a steadily growing community of researchers around the world motivated by the prospect of using cryo-EM to solve high-resolution structures of molecular targets associated with human diseases but have not been amenable to the traditionally employed X-ray crystallography and NMR spectroscopy. The mission of our Cryo-EM Facility is to provide access to cryo-EM instrumentation and to serve researchers from different backgrounds to tackle a wide variety of challenging biological structures ranging from individual molecules and their assemblies to subcellular organelles and whole cells that otherwise defy structural characterization.
The Cryo-EM Core Facility is equipped with high-end electron cryomicroscopes and the associated ancillary equipment for cryospecimen preparation, and is capable to provide atomic resolution imaging for single particle analysis (SPA cryo-EM) and for electron cryo-tomography (cryo-ET).
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