High-pressure freezing unit for visualizing cellular ultrastructure by electron microscopy

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To visualize cytoskeletal and membrane organization within cells, cryo-electron microscopy (cryoEM) provides the best resolution for visualizing the tiny components comprising the cellular ultrastructure. By snap freezing samples at extreme pressures, high-pressure freezing is designed to preserve complex cellular structures without introducing alterations or generating artifacts. CryoEM is essential for all "high-impact" molecular cell biology research, including that conducted by the ~30 member labs of the "Centre for Cell Biology, Development and Disease (C2D2)" at Simon Fraser University (SFU). At present, common use electron microscopes are available at the "SFU Advanced Functional Materials Research Institute (4D labs)," but they are configured and optimized for samples used in material sciences. To both facilitate use and make these electron microscopes more accessible for life science research, we request funds to acquire a high-pressure freezing system for cryoEM tissue/cell sample preparation. ******We propose to provide our institution with its first high-pressure freezing system to serve both the routine electron microscopy needs of all SFU researchers, but also to make cutting-edge cryoEM experiments possible (e.g. FIB-SEM). The Leica EM ICE offers ease of use, minimal support personnel for upkeep, and flexibility for cryoEM sample preparation with yeast, bacteria, and metazoan cells, as well as "soft matter" research. Because of its utility, there is overwhelming support for the acquisition of this system; there are many pledged contributions, which in addition to the NSERC RTI funds will be necessary to purchase the system; including four SFU Departments (MBB, BISC, BPK, Chem), the Faculty of Science, and a number of individual researchers (including the Lab Scientists' Committee at the Faculty of Health Science). This proposed system also permits future upgrades to the base system for investigating neuronal events that transpire a msec after electrical stimulation. ******The flexibility of the proposed microscope system matches the varied NSERC-funded research activities of the molecular cell biology labs at SFU. Leading this application, the Beh lab will use the proposed system to directly visualize the minuscule contact sites between intracellular membranes to understand the regulated assembly of membrane tethering complexes. The 10 co-applicants (Drs. T. Beischlag, N. Branda, M. Brockman, S. Gorski, N. Harden, H. Hutter, C. Krieger, M. Silverman, G. Tibbits, and E. Verheyen) will apply the new equipment to various research interests including nerve cell transport, viral factor-host membrane interactions, intercellular adhesion, and cellular etiologies of cancer. As an open resource to all SFU users, trained students will benefit from the mastery of this technologically advanced system for future employment in biotech.**