Extracellular flux analyzer to accurately monitor the real-time kinetics of cellular and mitochondrial bioenergetics

该申请寻求资金购买Seahorse XFe 24细胞外通量分析仪，这是一种能够以高通量24孔格式同时测量完整细胞或分离线粒体内多种代谢参数的最先进仪器。我们将使用这种仪器来研究所有细胞产生和消耗能量的代谢途径。这种分析对于西方基础生物学和临床科学研究的几个研究领域至关重要。Seahorse XFe分析仪通过测量完整活细胞中的耗氧量和细胞外酸化率，可以同时实时评估线粒体呼吸和细胞糖酵解。这种最先进的仪器目前还没有提供给西方研究人员。相反，我们依赖于过时的，低通量和耗时的技术来评估这些生物过程，因此我们的研究生产力是有限的。传统上，已经使用极谱（Clark型）或光纤（例如，Ocean Optics）氧电极系统来进行氧消耗的测量，所述氧电极系统一次只能读取一个或两个样品，并且在Western，这些系统的实际使用由于其低通量而受到严重限制。Seahorse系统具有优于这些常规系统的优点，因为其适于对粘附细胞以及细胞悬浮液或亚细胞组分进行实验，从而允许使用生理学相关模型。此外，该技术需要更少的细胞来测量氧消耗速率，并且细胞在分析后保持活力，用于后续实验或测定。Seahorse系统还配备了四个化合物注射端口，可编程顺序添加调节剂，用于能量通路询问、剂量反应或激动剂-拮抗剂分析。Seahorse XFe分析仪对于西方研究人员进行细胞能量代谢的高通量，准确，机械研究以及培养下一代科学家在尖端细胞和线粒体生物能量学方面将代谢分析与功能基因组和蛋白质组数据整合至关重要。

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.