Cryo-electron microscopy (cryo-EM) is an emerging technique in structural biology where the biological sample of interest is prepared under cryogenic conditions. The utility of cryo-electron microscopy stems from the fact that it allows the observation of specimens that have not been stained or fixed in any way, showing them in their "native" environment. This is in contrast to X-ray crystallography, which requires crystallizing the specimen, which can be difficult, and placing them in non-physiological environments, which can occasionally lead to functionally irrelevant conformational changes. The resolution of cryo-EM maps is improving steadily, and in recent years near-atomic resolution of some structures had been obtained, including those of viruses, ribosomes, mitochondrial protein complexes, ion channels, and enzyme complexes. The spiraling number of cryo-EM publications suggests this to be true: in 2016 alone, the technique has so far been used to map the structures of more than 800 molecules. Researchers can use cryo-EM to calculate the structure of a biological macromolecule or complex that has been flash-frozen in several conformations and so deduce the mechanism by which it works.
The course will cover the theory, practice and application of cryo-electron microscopy including single particle analysis, tomography and electron diffraction. Participants in the course will have supervised access to CSHL’s FEI Titan Krios and Falcon direct electron detector. This hands-on course will include lectures by leading experts who will discuss practical and conceptual approaches to structure determination using these techniques, as well as covering a wide range of state-of-the-art applications of cryo-EM in the biological sciences
Applications are open to individuals at universities and colleges, medical research institutions and industry, both from within the United States and from overseas, and to individuals at any stage in their postgraduate (PI, postdoc, grad student etc) career. Applicants must demonstrate that they are committed to applying cryo-electron microscopy directly to their own research, and that they work in an environment where such application is feasible and realistic.
Techniques to be Taught:
• Electron microscope alignment
• Negative stain and cryoEM sample preparation
• Automated Data Acquisition
• Single Particle Analysis
• Tomography data collection &analysis
• Atomic model building and validation