PHILIPS TECNAI 12 TRANSMISSION ELECTRON MICROSCOPE

此应用程序要求共享飞利浦Tecnai 12透射电子显微镜（TEM）。超微结构重建技术和免疫金定位方法正走向综合生物学的前沿。我们机构的研究人员正在使用这两种方法作为他们综合生物学研究的一部分。然而，由于我们机构缺乏冷冻电子显微镜（cryo-EM），他们的研究受到限制。另一个限制是无法获得传统的TEM。为我们的用户组选择最合适的基本仪器是基于Albertine博士作为电子显微镜专家的经验，特别是飞利浦的tem， Sundquist博士使用飞利浦冷冻电镜重建蛋白质组装的经验，以及来自这个国家使用冷冻电镜的专家晶体学家的建议。之所以选择飞利浦的TEM，是因为它同时满足了我们对低温电镜和常规电镜的需求。它的设计为两种EM模式提供了无与伦比的操作便利性，即使在安装冷冻阶段时也是如此。易于操作扩展到调查员的经验水平（从经验丰富的新手用户），包括我们有身体残疾的用户。我们的用户组有18名代表亨茨曼癌症研究所（HCI）的研究人员。我们的用户组将要求100%使用所请求的TEM。其他调查人员，包括年轻调查人员的需要，将通过持续提供仪器，包括晚上和周末来满足。我们的用户组有33个资助的公共卫生服务（PHS）研究资助和21个资助的非PHS研究资助，需要冷冻电镜和传统透射电镜。我们有16个未决的小灵通申请和3个未决的非小灵通拨款申请，建议使用这两种类型的TEM。首席研究员Albertine博士是电子显微镜专家，他将负责共享的TEM。与他一起工作的是一位全职的组织学/分子生物学技术人员；聘请一名全职电子显微镜技术人员（含承诺书）。内部咨询委员会(dr。Albertine， Sundquist和Fitzpatrick)将制定工具使用政策并解决使用冲突。机构资金承诺：我们有低温低温和柱下CCD相机系统的资金，这些系统在飞利浦Tecnai 12 TEM上有良好的记录，来自新的NIH RO1拨款和我们大学的配套资金（包括承诺信）。与此同时，我们也搬进了新的HCI研究大楼（1999年春天）。大约3000平方英尺的空间被设计用于显微镜。这个新空间将容纳所要求的TEM。此外，HCI承诺雇用一名全职的电磁技术人员来操作所要求的电磁设备并培训用户（包括承诺书）。服务合同资金由HCI（包括信件）承担，用于维护TEM，从02年开始（该年由制造商保修）。所要求的TEM将通过首次在我们的校园提供冷冻电镜，大大加强我们的用户组，HCI和健康科学中心社区的研究。

This application requests a shared Philips Tecnai 12 transmission electron microscope (TEM). Ultrastructural reconstruction techniques and immunogold localization methods are moving to the forefront of integrative biology. Investigators at our institution are using both approaches as part of their integrative biology studies. Their studies are limited, however, by the lack of a cryo-electron microscope (cryo-EM) at our institution. Another limitation is insufficient access to a conventional TEM. Selection of the most appropriate, basic instrument for our User Group is based on Dr. Albertine's experience as a electron microscopist in general and with Philips' TEMs in particular, Dr. Sundquist's experience with a Philips cryo-EM for reconstructions of protein assemblies, and advice from expert crystallographers in this country who use cryo-EM. Philips' TEM is selected because it simultaneously meets our needs for cryo-EM and conventional TEM. Its design provides unequaled ease of operation for both EM modes, even while the cryostage is installed. Ease of operation extends to the level of experience of investigators (from experienced to novice users), including our users who have physical disabilities. Our User Group has 18 investigators who represent the Huntsman Cancer Institute (HCI). Our User Group will require 100% of the usage of the requested TEM. Needs of other investigators, including young investigators, will be met by making the instrument available continuously, including evenings and weekends. Our User Group has 33 funded Public Health Service (PHS) research grants and 21 funded non-PHS research grants that require cryo-EM and conventional TEM. We have 16 pending PHS applications and 3 pending non-PHS grant applications that propose to use both types of TEM. Dr. Albertine, the Principal Investigator, is an expert electron microscopist who will be responsible for the shared TEM. Working with him is a full-time histology/molecular biology technician; a full-time electron microscopy technician will be hired (letter of commitment included). The Internal Advisory Committee (Drs. Albertine, Sundquist, and Fitzpatrick) will set instrument use policy and resolve use conflicts. Institutional funding commitment: We have funds for the cryostage and under-the-column CCD camera systems, which have a proven track record with the Philips Tecnai 12 TEM, from a new NIH RO1 grant and matching funds from our university (letters of commitment included). Coincident with this grant applicant is our move into the new HCI research building (spring of 1999). About 3,000 square feet of space is being designed for microscopy. This new space will accommodate the requested TEM. Furthermore, the HCI is committed to hire a full-time EM technician who will operate the requested TEM and train user users (letter of commitment included). Service contract funds are committed by the HCI (letter included) for maintaining the TEM, beginning year 02 (the year is covered by the manufacturer's warranty). The requested TEM will significantly enhance the research of our User Group, the HCI, and the Health Sciences Center community by providing, for the first time on our campus, cryo-EM.