CELL SORTER TO SUPPORT BASIC RESEARCH

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The applicants of this proposal conduct leading edge basic lipid research in fields of physiology, metabolism and cell signalling. Studies to date have addressed questions about how fatty acids, phospholipids, mechanical stress and exercise influence whole body physiology, nutrient metabolism and cell signalling. These fundamental questions have been addressed using a combination of whole body, cell and tissue culture approaches. The vast majority of tissues in the body are comprised of a wide range of cell types. This makes it difficult to determine which particular cells within a given tissue bed are responding to the intervention. Tissue analyses examining gene and protein changes make the basic assumption that all cells isolated are the same, however, this is a gross oversimplification and a multitude of other cell types are also present. For example, these may include stromal and epithelial cells or infiltrating immune cells. Centrifugation and density dependent methods are available but lack specificity and sensitivity, thus requiring large amounts of tissues (i.e. pooling organs from multiple animals) to obtain relatively impure preparations. Similarly, in cell culture approaches it is often desirable to sort cells based on phenotypical characteristics following treatment and then assess the desired experimental endpoints. These experimental needs can only be accomplished using a flow cytometer equipped with a cell sorter, infrastructure that is not readily available to the applicants. An instrument that meets the needs of the investigators and competitively priced is the Bio-Rad S3e cell sorter. This is an entry level workhorse flow cytometer which also has sorting capability. The only other direct competitor is the Sony SH800. However, while comparable in features, it is not competitively priced. Higher end flow cytometers have capabilities far exceeding the routine needs of these investigators and also costing upwards of 1 million. It is anticipated that acquiring this instrument will enhance the direct training of 25+ high quality personnel (per year) across 4 laboratories. Furthermore, it is anticipated that additional faculty and their trainees will seek the use of this instrument once established in the Department of Human Health and Nutritional Sciences at the University of Guelph. In recognition of the importance of this instrument, the Department will provide financial support to support budgetary needs. Overall, the ability to isolate purified populations of cells will enable the elucidation of mechanisms of action and enhance the training environment for many high quality personnel.