Grant: Research Tools and Instruments - Category 1 (<$150,000)

This application seeks funds to purchase a Seahorse XFe24 Extracellular Flux Analyzer, a state of the art instrument capable of simultaneously measuring multiple metabolic parameters within intact cells or isolated mitochondria in a high throughput 24-well format. We will use this instrument to study the metabolic pathways used by all cells to produce and consume energy. Such analyses are essential for several research areas in both fundamental biology and clinical science research at Western. The Seahorse XFe Analyzer permits simultaneous, real-time assessment of mitochondrial respiration and cellular glycolysis, by measuring oxygen consumption and extracellular acidification rates in intact live cells. This state-of-the-art instrument is not currently available to Western researchers. Instead we rely on dated, low throughput and time-consuming techniques to assess these biological processes, so our research productivity is limited. Traditionally, measurements of oxygen consumption have been undertaken using polarographic (Clark-type) or fiber optic (e.g. Ocean Optics) oxygen electrode systems, which can only read one or two samples at a time and, at Western, practical use of these systems is seriously limited due to their low throughput. The Seahorse system has an advantage over these conventional systems by being amenable to experiments on adherent cells, as well as cell suspensions or subcellular components, allowing use of physiologically relevant models. Moreover, the technology requires fewer cells for measuring oxygen consumption rates, and cells remain viable following analysis for use in subsequent experiments or assays. The Seahorse system also comes with four compound injection ports enabling programmed sequential addition of modulators for energy pathway interrogation, dose-response, or agonist-antagonist analyses. The Seahorse XFe Analyzer is critical for Western researchers to conduct high throughput, accurate, mechanistic investigations of cellular energy metabolism and to train the next generation of scientists in cutting edge cellular and mitochondrial bioenergetics to integrate metabolic profiling with their functional genomic and proteomic data.