PROJECT SUMMARY This proposal requests funds to acquire a transmission electron microscope (TEM) to support a diverse group of NIH-funded research projects at Vanderbilt University Medical Center and Vanderbilt University. The JEOL JEM 2100 Plus TEM identified in this grant is a modern 200 keV thermionic TEM. The TEM is equipped with a large format CMOS camera, automatic motorized apertures, turbo molecular vacuum system, and full integration SerialEM automation software. This TEM will replace the Cell Imaging Shared Resource’s (CISR) 20-year-old ThermoScientific Tecnai T12, a 120 keV LaB6 filament microscope. The Tecnai T12 has served the Vanderbilt community well throughout its lifetime facilitating numerous grants and studies, but its age has become an impediment to research progress with increasingly large downtimes, scarce availability of parts, and lack of modern features. Notably, this current Tecnai T12 TEM has been maintained by the CISR for almost its entire 20-year history. The 2100 Plus will not only serve as a suitable replacement for the Tecnai T12 but also add new capabilities identified as important to the Major User group. The 2100 Plus has tremendous flexibility, supporting accelerating voltages between 80-200 keV, allowing optimum imaging for small samples such as extracellular vesicles at lower voltage and up to 200 keV to image relatively thick sections for tomography. Acquiring high- quality tomograms of volumetric data will provide critical new capabilities increasingly required for state-of-the art analysis. The 2100 Plus’s stage, large format camera, and SerialEM automation software will also allow for efficient tiling of large areas at high-resolution; this type of data is becoming paramount for data quantification. Our Major Users will use the JEOL 2100 Plus to facilitate a wide range of NIH-funded research projects in cell, developmental, epithelial, and cancer biology. Projects include assembly of cell surface features (Goldenring, Tyska), metabolic phenotypes of mitochondria (Gannon, Rathmell, Skaar), extracellular matrix patterning (Kuchtey, Weaver), and cellular membrane trafficking (Weaver, Goldenring, Gannon). Together these investigations hold significance for our understanding of disease and illness related to birth defects, cancer, digestive disorders and many other human health problems. CISR will provide the infrastructure for introducing researchers across Vanderbilt to the newly available techniques, train users in operation of the instrument, and apply an established business model to support the long-term operation and maintenance of the system. In summary, acquisition of the JEOL 2100 Plus will allow for imaging capabilities at scales and spatial resolutions not currently available to the Vanderbilt community and will facilitate research on a diverse set of topics.

PROJECT NARRATIVE Transmission electron microscopes (TEMs) are essential instruments used to resolve cellular and macromolecular complexes beyond the diffraction limit of light. The JEOL JEM 2100 Plus TEM will support NIH- funded research into illnesses related to birth defects, digestive diseases, cancer, infectious disease, and other human health problems. There is a substantial and fundamental need for such an instrument to support the ongoing work of numerous NIH-sponsored investigators at Vanderbilt University Medical Center and Vanderbilt University, and significant resources are pledged to ensure successful implementation from both Vanderbilt University Medical Center and Vanderbilt University.