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.
