High Throughput Extracellular Flux Analyzer

描述（由申请人提供）： 在动物细胞中有两种途径产生大量的ATP-线粒体氧化磷酸化和糖酵解。这些ATP生成过程的调节是生物学家的根本兴趣，但这些过程很难在细胞系、原代细胞或组织样品中研究，特别是在适合高通量的96孔配置中。最近，Seahorse Bioscience开发了XF 24细胞外通量分析仪，可以连续测量在24孔板中培养的细胞的O2消耗和酸形成。通量分析仪使研究人员能够重复和非破坏性地测量氧化磷酸化，糖酵解，营养利用和单细胞培养中的其他过程。然而，24孔仪器不能提供足够的通量用于大型化学文库的初级筛选。由于原代细胞或组织需要在有限的时间内进行检查，因此使用这些样本的研究人员明显受限于可以对给定动物进行检查的变量数量。因此，我们的具体目标是：1）获得Seahorse 96孔细胞外通量分析仪，用于测量O2吸收和酸释放，2）将该仪器置于多用户环境中，提供技术支持，用户培训和应用专业知识，以支持单个用户项目。这项新兴技术将独特地允许NIH资助的研究人员从事各种项目，以评估正常生理学和疾病模型中的生物能量学和代谢反应，包括癌症，酒精性肝毒性，黄斑变性，心脏缺血和衰竭，多器官衰竭和糖尿病。这些研究的结果可能导致为其中一些疾病开发新的治疗方法。

DESCRIPTION (provided by applicant): Two pathways generate the bulk of ATP in animal cells -mitochondrial oxidative phosphorylation and glycolysis. Regulation of these ATP-generating processes is of fundamental interest to biologists, but these processes are difficult to study in cell lines, primary cells or tissue samples, especially in a 96-well configuration amenable to high throughput. Recently, Seahorse Bioscience has developed the XF24 Extracellular Flux Analyzer that permits serial measurements of O2 consumption and acid formation of cells cultured in 24-well plates. The flux analyzer enabled investigators to measure repetitively and non-destructively oxidative phosphorylation, glycolysis, nutrient utilization and other processes in single cell cultures. However, the 24-well instrument does not offer sufficient throughput for primary screening of large chemical libraries. Because primary cells or tissues need to examined in a limited time period, investigators using these samples are significantly limited to the number of variables that can be examined for a given animal. Accordingly, our specific aims are to 1) acquire a Seahorse 96-well Extracellular Flux Analyzer for measurement of O2 uptake and acid release, and 2) place this instrument in a multi-user environment that provides technical support, user training and applications expertise in support of individual user projects. This emerging technology will uniquely permit NIH-funded researchers working on a variety of projects to assess bioenergetics and metabolic responses in normal physiology and models of disease, including cancer, alcoholic hepatotoxicity, macular degeneration, heart ischemia and failure, multiple organ failure and diabetes. The results of those studies could lead to the development of new therapeutic treatments for some of these diseases.