1. Project Summary and Abstract Cryogenic-electron microscopy (cryo-EM) combined with Single Particle Analysis (SPA) have forever changed life-science research. Undoubtedly, we have entered the post-crystallography era of structural biology. However, a drawback of cryo-EM that limits its applicability is the cost of the instrumentation required for high- resolution data collection. A state-of-the-art Krios 300 kV electron microscope is around 5-7 million USD that quickly reaches and often exceeds 10 million USD with added costs for renovation, service contract, and supportive staff. This is not only unaffordable for many universities but also insufficient to establish cryo-EM as a routine methodology to study biological macromolecules. A considerable part of the cryo-EM SPA pipeline is spent screening specimens, which includes identifying vitrification conditions and imaging grids. Such an essential yet time-consuming prep-work can be achieved effectively and perhaps even more efficiently on less- powerful 200 kV microscopes that are easier to maintain and can be operated by non-experienced users. Therefore, as part of a cost-sharing plan to establish a state-of-the-art center for electron microscopy at Thomas Jefferson University (TJU), we request funds to purchase a Thermo Scientific™ Glacios™ Cryo- Transmission Electron Microscope (cryo-TEM) equipped with Falcon 4 Direct Electron Detector. We have identified a group of 11 major core users from both inside TJU and in nearby institutions who eagerly support this proposal. As proof of commitment to establish cryo-EM at TJU, the Provost, Dr. Mark Tykocinski has agreed to lease the Glacios at zero money down for 12 months with no obligation to buy the instrument. After the completion of the 12 months, the University can terminate the lease and return the microscope or extend the lease by making three annual installments or purchase the equipment for cash at the fair market value. The University has also committed to infusing additional funds to support a yearly service contract, equipment maintenance fees, and support a recently recruited cryo-EM manager. If funded, this S10 will help defray a significant portion of the cost of the microscope, providing vital economic assistance to the University at a time of considerable financial uncertainty. We have opted for the Thermo Scientific™ Glacios™ 200 kV cryo-TEM with Falcon 4 camera, which is an affordable workhorse that provides an excellent setup for in house cryo-EM. Also, in light of the significant investment made by NIH to develop synchrotron-like cryo-EM federal facilities around the US, we believe that acquiring a screening cryo-TEM is a sensible investment for a metropolitan campus. The Glacios expands and enhances the quality, capability, efficiency, and cost-effectiveness of cryo-EM studies for our researchers and those at local and regional universities and research centers in the Philadelphia area.