Instrumentation Upgrade: acquisition of an intermediate voltage TEM

在这里，我们寻求资金购买一台现代化的中压（300千伏）透射电子显微镜（F30 Polara）。该显微镜将取代斯坦福-伯纳姆医学研究所（SBMRI）现有的FEI Tecnai F20。F30将集成到现有的由低温tem和辅助设备组成的单元中。它将通过半自动化的样品处理和更强大的筛选能力、更好更强大的电子光学、LN2稳定阶段和更高的倾斜系列完成率，在LN2温度下提供更强大、高质量/高通量的低温层析成像数据采集手段。总之，这一升级将使我们能够优化来自我们科学团队的广泛生物样品的数据质量和分辨率，使我们能够将分子机制和功能输出联系起来，提供疾病的规模集成模型，并为新疗法的开发提供新的视角。拟议的仪器升级将支持SBMRI的科学家团队和TSRI、UCSD、UCLA、耶鲁大学、弗吉尼亚大学医学院、哈佛医学院、Stowers医学研究所和新墨西哥大学健康科学中心的研究人员。所有这些研究人员都是通过美国国立卫生研究院的项目拨款来资助的，他们在发育生物学、炎症、先天免疫、骨矿化、转移和传染病等领域进行广泛的健康相关研究。所有在这个应用中突出的项目都使用了低温层析成像，从单个大分子组装结构的层析成像到原位全细胞层析成像。这些项目只是潜在的更大项目池的代表，这些项目可以从使用这种专用的现代冷冻断层扫描F30范围中受益匪浅。

DESCRIPTION (provided by applicant): PROJECT SUMMARY Here, we seek funding to purchase a modern, intermediate voltage (300 kV) transmission electron microscope (F30 Polara). This microscope will replace the existing FEI Tecnai F20 at Sanford-Burnham Medical Research Institute (SBMRI). The F30 will be integrated into an existing unit that comprises of cryo-TEMs and ancillary equipment. It will provide the means for a more robust, high-quality/high-throughput cryo-tomography data acquisition at LN2 temperatures by semi-automation of sample handling and more robust screening capabilities, better and more powerful electron optics, LN2 stable stage and higher rates of tilt series completion. Together, this upgrade will allow us to optimize data quality and resolution for a broad range of biological samples from our scientific teams, enabling us to correlate molecular mechanisms and functional outputs, to provide scale-integrated models of disease, and to provide new perspectives for the development of new therapeutics. The proposed instrumentation upgrade will support teams of scientists from SBMRI and researchers from TSRI, UCSD, UCLA, Yale University, University of Virginia School of Medicine, Harvard Medical School, Stowers Institute for Medical Research, and the University of New Mexico Health Sciences Center. All these researchers are funded through project grants from the NIH to pursue a broad range of health-related studies in the areas of developmental biology, inflammation, innate immunity, bone mineralization, metastasis, and infectious diseases. All of the projects highlighted in this application employ cryo-tomography imaging, ranging from tomography of single macromolecular assembly structure determination to in-situ whole cell tomography. These projects are only representatives of a potentially much larger pool of projects that could greatly benefit from access to this dedicated, modern cryo-tomography F30 scope.