The goal of this application is to obtain funding for a Seahorse XF96 (extracellular flux, 96 well) analyzer. A resurgence of interest in cell metabolism and energy use has recently impacted almost all areas of research on cell biology. This includes research into metabolic, neurodegenerative and cardiovascular diseases, cancer, drug safety, basic cell physiology, immunology and the processes of inflammation. The importance of this analysis among various disciplines is mirrored by the range in research interests of the investigators joining together for this S10 equipment grant application. Much of this renewed interest has been driven by the various """"""""omic"""""""" analyses, which often identify variations in energy use among cells in both animal models and patient samples. Seahorse Bioscience has met this renewed interest and generated a series of machines that greatly improve the throughput and ease of use. Their instruments are perfect for a core facility as users can be rapidly trained to analyze and interrogate their experimental samples of interest. The XF96 is capable of measuring both glycolysis and oxidative phosphorylation simultaneously on primary cells or isolated mitochondria continuously in real time. The 96-well microplate format allows for smaller cell sample sizes to measure multiple analytes (currently oxygen and protons) in real time on adherent and suspension cells in situ. In addition, up to four test compounds (drugs or substrates or agonists or antagonists) may be added automatically to each well with measurements performed before and after addition of each reagent. Lastly, this technology fills a critical niche currently absent from most research labs by opening up new insights and paradigms of activation in cell physiology and disease states.
|Shorter, Shayla K; Schnell, Frederick J; McMaster, Sean R et al. (2016) Viral Escape Mutant Epitope Maintains TCR Affinity for Antigen yet Curtails CD8 T Cell Responses. PLoS One 11:e0149582|