MRI: Acquisition of a Transmission Electron Microscope and Digital Imaging: Research and Research Training in an Undergraduate College Environment

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A grant has been awarded to Dr. Jan Robert Factor at Purchase College, State University of New York, to acquire a digital transmission electron microscope (TEM) and related equipment for digital image processing and microscopical image analysis. Specifically, a TEM will be acquired; an image analysis system will be upgraded; as well as an ultramicrotome and diamond knife for preparing biological samples for TEM. The TEM and related instruments will be used to carry out several research projects and research training in an undergraduate college environment. This equipment will enable research projects in the areas of cellular immune defenses in the lobster (Homarus americanus); digestive tissues of the lobster (Homarus americanus); characterization of the cell surface of Renibacterium salmoninarum; uptake and growth of Renibacterium salmoninarum in trout macrophage; cellular events induced by fatty acids.The following research projects will be carried out. 1. Cellular Immune Defenses in the Lobster, Homarus americanus, Jan Robert Factor. Removal of foreign particles from the blood of the lobster by a large population of fixed phagocytes will be studied using fluorescence light microscopy and analysis of digital images, to determine if adverse environmental conditions and anthropogenic toxins cause disease and mass mortalities. TEM will be used to elucidate the unusual mechanism of phagocytic uptake by the fixed phagocytes. 2. Digestive Tissues of the Lobster, Homarus americanus, Jan Robert Factor. Understanding of digestive processes in this ecologically and economically important species has been reliant upon electron-microscopical observations, and further progress in understanding digestive tissues absolutely requires the use of transmission electron microscopy. Studies will examine cells and tissues of the tegumental glands, complex basement membranes, connective-tissue fibers, and tissue organization. 3. Characterization of the Cell Surface of Renibacterium salmoninarum: Fimbriae and p57 Surface Proteins, James G. Daly. R. salmoninarum is a unique intracellular pathogen that causes bacterial kidney disease in salmonid fishes. Its major cell surface antigen (p57) may be important to pathogenesis. p57 and surface fimbriae may be the same, however, p57 antibody fails to attach to the fimbriae. Transmission electron microscopy will be used to examine the cell surface to determine whether p57 is indeed a fimbria or whether they are distinctive proteins. 4. Uptake and Growth of Renibacterium salmoninarum in Trout Macrophage, James G. Daly. This project will use both in vitro and in vivo infected macrophages to follow the infection and survival of R. salmoninarum within trout macrophage. 5. Ultrastructural Localization of Cellular Events Induced by Fatty Acids, Joanne Kivela Tillotson. Biochemical pathways of growth factor and eicosanoid signaling will be localized by TEM in order to define the influence of specific dietary fatty acid administration upon cell proliferation, and the effects of the fatty acids on programmed cell death. Combined with cell culture and biochemical studies, this will further our understanding of the role of exogenous factors in cell proliferation. This equipment will significantly enhance the research capabilities at Purchase College and enable faculty research that is not now possible, with an important impact on the research of more than half of the Biology faculty. The research includes several areas of practical scientific significance: disease defenses and processes in two commercially important groups of animals (the American lobster and salmonid fishes), and the influence of dietary fatty acids on cell proliferation and programmed cell death. Undergraduate students will be trained in the required techniques in two courses, providing the tools needed to participate in the faculty research projects. The requested equipment will also be used to encourage students to pursue science careers through a variety of local school programs.