A National Resource for computer-aided intermediate voltage transmission of electron microscopy (IVEM) and 3-dimensional (3-D) image analysis had been established to serve the Southwest region of the country. these facilities are available to investigators working on biological problems that could benefit from the use of thicker specimens and 3-D image analysis. We have directed our plan for continuation of the core technological research toward the development of technologies that will contribute to structural neurobiology as well as cell biology anmolecular biology. Our goals for collaboration, service, training and dissemination are all related to expanding the use of these technologies to maximize their value to the biomedical community. We plan to investigate the application of a variety of selective contrast enhancing methods to model/system biological problems that will clearly benefit from the unique capabilities of a computer-coupled IVEM. Our Core projects include development of labeling procedures enabling correlation of light and IVEM analyse of specimens to examine the architecture of neuronal systems ad the dynamics of subcellular processes, new IVEM compatible macromolecular probes, methods to improve penetration of probes into frozen sections, improved selective staining of cell and subcellular structure. The capabilities to be developed and research accomplishments anticipated in all of the Core projects areas involving methods of specimens preparations will be augmented by available and planned features of the Resource image processing facilities. We emphasize the use of computer-aided methods for the enhancement of image contrast, extraction of accurate 3-D information from within single thick specimens and the 3-D reconstruction of larger structural complexes using tomographic and serial thick section analysis. We are developing a serial tomography method in which information within thick sections obtained by tomography may be combined across a series section. Ongoing projects will expand our capability to analyze large 3-D data sets by distributing computation to supercomputers linked to the Resource via high-speed networks. We are exploring the feasibility of remote interaction with the IVEM to control the microscope to acquire and view data by linking the remotely located researcher to the microscope via the network; thereby providing researchers greater access to the Resource. Other projects will provide: user interactive modification of operating conditions of the IVEM in order to alter contrast; microscope-coupled image processing and analysis; and semi-automated 3-D image acquisition and reconstitution. Training of users in the application of technology will occur at the Resource. In addition, workshops and various other forms of scientific communication will be used to disseminate the Resource technologies.
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