Recent technical innovations in light and electron microscopy, the development of new cellular markers, an increased capacity to generate antibodies to specific cellular antigens, and improved techniques for in situ hybridization of genetic probes have enormously enhanced our ability to study structures at the cellular molecular levels. The investigators will implement instrumentation which combines these recent advanced anatomical techniques with methods for computer image analysis that they have developed over the past twenty years. The instrumentation will be used for visualization in three dimensions (3D) of cells, subcellular structures and specific molecular markers within cells and tissues. Computerized image processing will be used to display stereo pairs or images with real-time rotation for 3D visualization, with color used to distinguish the different markers or structures. The instrumentation will be used for many different research projects. These include studies of the locations of viral proteins in infected cells, the distribution of mRNAs in mouse cerebellar neurons, the spatiotemporal pattern of expression of homeobox-containing genes, the morphology and interactions of developing and regenerating neurons, the distribution of different types of vesicles during exocytosis, and the assembly and transport of histones in yeast, as well as several others. The proposed instrumentation will bring together neurobiologists and molecular biologists with widely differing scientific backgrounds but with a common interest, and will thus serve as a focus for the exchange of ideas and technical expertise, as well as a means for initiating cross-disciplinary collaborations. It will play an important role in expanding the range of approaches used by individual investigators, effectively helping the neurobiologists and cell biologists to become more molecularly oriented, and the molecular biologists more cellularly oriented.
