The objective of this laboratory is to develop techniques for quantitative 3D light and electron microscopy and to make these available to the scientific community for research and education. A 400 kV transmission electron microscope allows examination of specimens 10-20 times thicker than usual and has been modified for an unlimited range of specimen tilt angles for 3D imaging. A laser scanning confocal light microscope provides optical sectioning in thick specimens, including wet and living specimens, superior to that obtainable using ordinary light microscopes. The foci in the core research are quantitation, comparison of images using different imaging modalities, and the evaluation of the effects of preparation methods and other potential sources of artifacts. Images are processed using image analysis programs imported from other sources or developed here. Programs are available for reconstruction from serial section images and from images taken at various tilts (including stereo images). These programs provide 3D localization of particulate markers (gold particles), reconstruction of membranes and other surfaces, and quantitative analysis of networks. Initially, manual processes are implemented. We plan in the longer term to automate as many processes as possible. Current collaborative research projects are focussed on the localization of surfaces and surface receptor molecules in motile cells, the organization of the cytoplasmic matrix, the structure of protein networks, the structure of the transverse tubular system in skeletal muscle, and the development of the fibrillar system in differentiating cardiac and skeletal muscle cells. Service projects cover a broad range of topics. Users have come from several surrounding states and from Japan. Training is done on-site at the facility, and through workshops presented at the facility and elsewhere. Dissemination of information on the facility is done through publications, lectures and, posters presented at national and international meetings, and direct mailing to biomedical scientists.
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