Acquisition of a High Pressure Rapid Freezer

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The High Pressure Rapid Freezer (HPRF) will be used to freeze samples of relatively large dimensions without the formation of large ice crystals, making it feasible to study complex cellular systems, tissues, using rapid cryofixation. This opens new frontiers in the exploration of rapidly fixed eukaryotic organisms. Coupled with freeze substitution, rapid freezing will allow study of labile structures, short lived events, and other advances over conventional preparations already shown when single cell suspensions have been studied using this approach. Projects will use the superior characteristics of freeze substituted tissues prepared with the HPRF to investigate short lived events (neural tissue alterations due to ion concentrations, secretion in salivary glands of ticks, secretion of calcium in plant cells), labile structures (oocyte cytoskeleton, cotton fiber cytoskeleton), osmotic-sensitive structures (mitochondria in chemotherapy, symbiotic interfaces), immunolocalization of various antigens (cytskeleton, cytokinin, anticancer agents), and symbiology between dinoflagellates and sea anemone, and plants and nitrogen-fixing bacteria. The project involves research in several regional institutions, and will examine plant, animal, and microbial systems that have been recalcitrant to cryopreservation using previously available equipment. The instrument will be the only one in the southeastern United States, and would be made available to qualified regional users.