PHILIPS CM120 TRANSMISSION ELECTRON MICROSCOPE

透射电子显微镜的一个重要进展是 近年来，用于观察样品的方法的发展 嵌在非结晶（玻璃状）冰中。这些样本 尽可能靠近原位，以便通过电子进行观察 显微镜请求为菲利普斯CM 120变速箱EM提供资金 配备冷冻EM操作，以便这项技术可以很容易地 可供加州大学伯克利分校的研究人员使用， 旧金山弗朗西斯科湾区。我们建议使用冷冻电镜研究囊泡 S.酿酒酵母分泌途径，脂质体，梁- 用于骨置换手术的敏感人造聚合物， 结核分枝杆菌，纳米晶体，以及一些不同的二维 蛋白质晶体此外，我们还有一些核心用户， 在传统模式下使用显微镜，但谁将受益于 现代显微镜的分辨率更高， 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. cerevisiae和S. pombe酵母、果蝇和C.优雅下 材料将通过高压冷冻和冷冻制备 以最佳保存形态和抗原性。 该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，一个收费服务设施开放， 所有校园用户以及周边的研究人员 机构，如加州大学旧金山分校弗朗西斯科，加州大学戴维斯分校，加州大学圣克鲁斯和 斯坦福大学。据我们所知这些学校都没有冷冻电镜 可供一般使用。本提案中的所有主要用途均为 经验丰富的EM用户，大多数都有NIH的资助。选定的核心用户 电磁实验室主管将组成一个咨询委员会， 监督工具的使用并制定政策。其他区域使用 研究人员将在核心研究结束后的剩余时间内 调查员使用。加州大学伯克利分校将提供空间和工作人员， 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.