PHILIPS CM120 TRANSMISSION ELECTRON MICROSCOPE

透射式电子显微镜的重要进展之一 近几年来，观察样品的方法得到了发展 嵌在非晶态(玻璃)冰中。这些样品大约是 尽可能靠近自然位置，以便用电子进行观察 显微镜。请为Phillips CM120变速器EM申请资金 配备了低温EM操作，因此这项技术可以很容易地 加州大学伯克利分校的研究人员和 旧金山湾区。我们建议使用低温电子显微镜来研究水泡 酿酒酵母分泌途径的组分，脂质体，BEAM- 骨替代手术中使用的敏感人造聚合物， 结核分枝杆菌、纳米晶体和一些不同的二维 蛋白质晶体。此外，我们拥有许多核心用户，他们将 在传统模式下使用显微镜，但谁将受益于 拥有分辨率更高、分辨率更高的现代显微镜 加速电压。该小组主要对使用 用于EM免疫定位工作和/或表征突变体的显微镜 硬骨鱼视网膜、哺乳动物胃细胞、S。 酿酒酵母、金黄色葡萄球菌、果蝇和线虫。较低的 材料将通过高压冷冻和冷冻来制备 替代了形态和抗原性的最佳保存。 显微镜将被安置在电子显微镜实验室。 加州大学伯克利分校，收费服务设施开放 给所有校园用户以及来自周围的研究人员 加州大学旧金山分校、加州大学戴维斯分校、加州大学圣克鲁斯分校和 斯坦福大学。据我们所知，这些学校都没有冷冻EM 可供一般使用。这项提案的所有主要用途是 经验丰富的新兴市场用户和大多数都有NIH的资助。精选核心用户加 EM实验室的监管者将组成一个咨询委员会，以 监督工具的使用并制定政策。由其他地区使用 调查人员将在核心结束后的剩余时间内受到鼓励 调查员使用。加州大学伯克利分校将为运营提供空间和人员 显微镜以及40,000美元的延长服务合同。

One of the important advances in transmission electron microscopy in recent years has been development of methods for observing samples embedded in non-crystalline (vitreous) ice. These samples are about as near to the native site as possible for observation by electron microscopy. Funds are requested for a Phillips CM120 Transmission EM equipped for cryoEM operation so that this technology can be readily available to researchers at the University of California, Berkeley and the San Francisco Bay Area. We propose to use cryoEM to study vesicle fractions from the S. cerevisiae secretory pathway, liposomes, beam- sensitive artificial polymers used in bone-replacement surgery, Mycobacterium tuberculosis, nano-crystals, and a number of different 2-D protein crystals. In addition, we have a number of core users who will use the microscope in conventional mode, but who will benefit from having a modern microscope with improved resolution and higher accelerating voltage. This group is primarily interested in using the microscope for EM immunolocalization work and/or characterizing mutant phenotypes in teleost fish retina, mammalian stomach cells, S. cerevisiae and S. pombe yeasts, Drosophila and C. elegans. The lower material will be prepared by high pressure freezing and freeze substitution for optimum preservation of morphology and antigenicity. The microscope will be housed in the Electron Microscope Laboratory at the University of California, Berkeley, a fee-for-service facility open to all campus users as well as researchers from the surrounding institutions such as UC San Francisco, UC Davis, UC Santa Cruz and Stanford. As far as we known, none of these schools has a cryoEM available for general use. All of the major uses in this proposal are experienced EM users and most have NIH funding. Selected core users plus the Supervisor of the EM Lab will make up an advisory committee to oversee use of the instrument and set policy. Use by other regional investigators will be encouraged in the time remaining after core investigator use. UC Berkeley will provide space and staff for operating the microscope as well as $40,000 for an extended service contract.