Acquisition of 200 KeV Transmission Electron Microscope

本提案的目的是为购买一台飞利浦CM- 200透射电子显微镜申请资金，该显微镜将被安置在伊利诺伊大学校园贝克曼研究所的可视化设施（BVF）中。首席研究员将使用这台电子显微镜进行各种研究，包括(i)神经可塑性及其与潜在细胞和系统水平过程的关系（William Greenough）， （ii）单分散棒圈嵌段共聚物的研究（Samuel Stupp）， （iii）核和染色体结构分析（Andrew Belmont）， （iv）大脑组织和行为功能的性别差异（Janice Juraska），(v)运动功能障碍遗传动物模型中小脑神经回路形态和功能异常的表征（Louise Abbott）。除了这些核心项目之外，还有6个来自不同学科的其他研究项目，该工具为这些项目提供了直接或长期的好处。获得新仪器的目的是双重的。首先，它将为UIUC校园内的生物研究提供中压仪器。该地区唯一的中压仪器专门用于材料科学研究和需要更高电压的项目的生物学家必须前往偏远地区。其次，它将取代目前位于BVF的已有18年历史的仪器，该仪器日益受到与年龄有关的问题的困扰。除了提供成像较厚的标本的独特能力，这种显微镜将有许多其他重要的优势超过现有的仪器。通过计算机接口控制仪器和使用CCD相机收集显微照片的能力将大有帮助，并且在某些情况下对几种应用是必不可少的。具体来说，对于串行切片、定量立体成像和单轴断层扫描等涉及收集或检查大量图像的技术，可以设计半自动采集程序，不仅简化了过程，而且确保了电子剂量最小化和感兴趣的区域最大化。此外，这些三维重建技术的某些应用实际上需要每个数据集数百张图像，这些实际考虑的应用只有在直接数字数据采集的可用性下才可行。这些控制和成像能力也将大大提高常规高质量图像的获取。这种新型显微镜的进一步优点源于过去20年来技术的进步。这导致了校准程序的简化，光束相干性、透镜和测角仪阶段的改进，以及在低剂量下获取图像以限制光束损害的技术的实施。该显微镜将位于贝克曼可视化设施内，该设施内有许多其他相关仪器（包括共聚焦显微镜，立体工作站和各种光学显微镜）以及配套设备（湿实验室，显微切片机，暗室）和用于图像数字处理和分析的优秀设施。该显微镜位于贝克曼研究所的多学科环境中，该研究所还包括国家超级计算应用中心的一部分，为利用一些最新技术进行电子显微照片的在线处理和分析提供了一个极好的环境。我们坚定地致力于向校园范围内的用户群体提供使用该仪器及其相关的特殊设施的机会，这些用户群体的研究全部或部分依赖于电子显微镜的应用。贝克曼学院和整个大学对这个项目都有热情的承诺。贝克曼研究所可视化设施为该项目提供了50%的配套资金，并为设备提供了配套基础设施，这证明了这一点。

The purpose of this proposal is to request funds for the purchase of a Philips CM- 200 Transmission Electron Microscope to be housed in the Visualization Facility (BVF) of the Beckman Institute on the University of Illinois campus. The Principal Investigators will be using this electron microscope for a variety of studies including (i) neuroplasticity and its relationship to underlying cellular and system level processes (William Greenough), (ii) the study of monodisperse rod-coil block copolymers (Samuel Stupp), (iii) analysis of nuclear and chromosome architecture (Andrew Belmont), (iv) sex differences in the organization and behavioral function of the brain (Janice Juraska), and (v) the characterization of morphological and functional abnormalities in cerebellar neural circuitry in a genetic animal model of movement dysfunction (Louise Abbott). In addition to these core projects there are 6 other research projects from a variety of disciplines for which this instrument provides immediate or long term benefits. The purpose of obtaining a new instrument is twofold. Firstly it will provide access to an intermediate voltage instrument for biological research on the UIUC campus. The only intermediate voltage instruments in this region are exclusively devoted to material science research and biologists with projects for which higher voltages are necessary have to travel to remote sites. Secondly it will replace an existing 18 year old instrument currently located in the BVF that is suffering increasingly from age related problems. In addition to providing unique capabilities for imaging thicker specimens this microscope will have a number of other important advantages over the existing instrument. The ability to control the instrument through a computer interface and to collect micrographs using a CCD camera will greatly aid, and in certain cases be essential for several applications. Specifically, for techniques such as serial sectioning, quantitative stereology and single axis tomography which involve the collection or examination of large numbers of images, semi-automatic acquisition procedures can be devised that not only simplify the process but also ensure that the electron dose is minimized and the region of interest is maximized. Furthermore, certain applications of these 3-dimensional reconstruction techniques require literally hundreds of images per data set and these applications for practical considerations will only be feasible given the availability of direct digital data acquisition. These control and imaging capabilities will also greatly improve the routine acquisition of high quality images. Further advantages of the new microscope arise from the advances in technology over the past 20 years. These have led to simplification of alignment procedures, improvements in beam coherence, lenses and goniometer stages and the implementation of techniques for acquiring images at low doses in order to limit beam damage. The microscope will be located in the Beckman Visualization Facility which houses a number of other related instruments (including a confocal microscope, a stereology workstation and various light microscopes) as well as supporting equipment (wet lab, microtomes, darkrooms) and excellent facilities for the digital processing and analysis of images. The location of the microscope in the multi- disciplinary environment of the Beckman Institute, which also includes part of the National Center for Supercomputing Applications, provides an excellent environment to take advantage of some of the latest technology available for the on- line processing and analysis of electron micrographs. We are strongly committed to providing access to this instrument and its associated, special facilities to the campus wide user group whose research wholly or partly depends on the application of electron microscopy. There is enthusiastic commitment to this project from both the Beckman In stitute and the University as a whole. This has been demonstrated by the contribution of 50% matching funds for the project as well as the supporting infrastructure for the equipment provided by the Beckman Institute Visualization Facility.