ABSTRACT Proposed major research instrumentation This proposal is a request for an imaging -- recording center which will enhance the capabilities of an existing confocal microscope, replace an out-of-date image analysis system, and introduce techniques such as time lapse micrography that are not currently available at the Medical College of Pennsylvania. The major optical components of the system consist of an ultraviolet laser for a Biorad MRC-600 confocal microscope, an inverted Zeiss microscope, cooled CCD camera, a gel analysis system, and various analysis, storage, and output devices. A complete system for several electrophysiological recording techniques will be incorporated into the light microscope setup. This will include facilities for whole cell and single channel patch clamping, sharp microelectrode work, and microinjection of cells for molecular biology manipulations. The resulting system will add a state-of-the- art facility for the combined imaging and recording of cells which will greatly extend the scope of the investigators to pursue studies of cell function. Type of research/research training conducted The interests of all of the investigators represented in this proposal center around issues of cell function including intracellular signaling, regulated exocytosis, and mechanisms of development. While several individuals study neuronal preparations, we feel strongly that the major questions in cell biology will be answered by studies of many cell types and should be approached by a broad spectrum of techniques. Technical expertise within the group in this proposal includes physiology, biophysics, imaging, and molecular biology. The investigators are all supported by funds from government and private sources. The existence of a multi- disciplinary, multi-technique center will greatly benefit both the investigators and training at all levels from graduate students through advanced post-doctoral fellows. Activity th at would result if funded The research community at the Medical College of Pennsylvania is a relatively small but highly interactive and productive group. The equipment requested is too expensive to be justified by individual grants. The existence of an imaging - recording center will both extend and enhance the research programs of individual investigators, and will increase the amount of interdisciplinary interaction. Several of the individuals listed in this proposal participate in an interdepartmental "Secretion Journal Club". If this research is funded we would extend the scope of the Secretary club to include other aspects of cell development and communication. A major benefit of such a center is that imaging and electrophysiology will be combined in one location, lowering the barriers for individuals to perform experiments using unfamiliar techniques. Although the imaging center will be administered by the Department of Anatomy and Neurobiology, it will be made readily available to other individuals and departments. The presence of the confocal microscope during the last year and a half has motivated several interdepartmental collaborations. Enhancement of imaging and recording resources will greatly expand the number and strength of such collaborations.
