Seahorse Bioscience Extracellular Flux Analyzer

描述（由申请人提供）：申请资金购买Seahorse Bioscience XF 96细胞外通量分析仪酶标仪。该仪器允许测量两个基本的重要参数，O2和pH值，用于评估细胞或分离的线粒体的生物能量分布。该装置允许以96孔读板器格式同时连续记录时间过程，具有自动注射两种不同化合物的能力，允许测定O2消耗速率（即，线粒体氧化磷酸化或OxPhos的速率）和细胞外酸中毒（指示通过厌氧糖酵解产生乳酸的速率）。此外，该设备能够使用非常少量的样品量测量分离的线粒体悬浮液中的O2消耗，这是我们开发的一种方法，并在初步实验中使用从Seahorse Biosciences借来的XF-96仪器进行验证。通过设计适当的方案，已经证明了来自不同实验组的各种细胞类型和线粒体的生物能量谱，并且可以在高通量平台上鉴定底物选择、代谢控制位点和代谢缺陷的差异。 该仪器加快能量代谢研究步伐的独特能力促使我们要求将该设备作为生物能量分析核心的基石，该核心将补充约翰霍普金斯大学的细胞和组织水平成像平台，并为各种经验丰富的研究人员提供服务，这些研究人员是心血管生物学，癌症生物学和衰老等多个学科的领导者。七个主要用户，他们都领导着NIH资助的多研究者，多学科研究项目，形成了核心用户组，其他几个次要用户已经在演示单元上的试运行中生成数据。建立该中心后，预计还将建立广泛的用户基础，因为该大学有许多具有类似研究兴趣的研究人员。该技术是经过验证的，并已自2006年以来，然而，这将是第一个这样的仪器安装在校园作为核心资源，并提供给整个霍普金斯研究界。 细胞的生物能谱的高通量功能表征是一个未满足的需求，其已经落后于由基因组学、蛋白质组学和代谢组学产生的信息爆炸。因此，了解参与能量代谢的蛋白质中越来越多的疾病相关变化的作用的唯一希望将是用Seahorse Bioscience仪器实现的多路时间分辨记录取代繁琐耗时的经典呼吸或代谢物测定。

DESCRIPTION (provided by applicant): Funds are requested to purchase a Seahorse Bioscience XF96 extracellular flux analyzer platereader. The instrument permits the measurement of two fundamentally important parameters, O2 and pH, for assessing the bioenergetic profile of cells or isolated mitochondria. The device permits sequential time course recording simultaneously in a 96-well platereader format with the ability to inject two different compounds automatically, allowing the determination of the rates of O2 consumption (i.e., the rate of mitochondrial oxidative phosphorylation or OxPhos) and extracellular acidosis (indicative of the rate of lactate production by anaerobic glycolysis). Moreover, the device is capable of measuring O2 consumption in suspensions of isolated mitochondria using very small sample amounts, a method we developed and validated in preliminary experiments using an XF-96 instrument on loan from Seahorse Biosciences. With the design of appropriate protocols, the bioenergetic profile of a variety of cell types and mitochondria from different experimental groups has been demonstrated, and differences in substrate selection, sites of metabolic control, and defects in metabolism can be identified on a high throughput platform. The unique ability of this instrument to accelerate the pace of research on energy metabolism has motivated us to request the device as the cornerstone of a Bioenergetic Profiling Core that will complement cellular and tissue level imaging platforms at Johns Hopkins University and serve a variety of highly experienced investigators that are leaders in several disciplines including cardiovascular biology, cancer biology, and aging. Seven major users, who all lead NIH-funded multi-investigator, multidisciplinary research programs, form the core user group, and several other minor users have already been generating data in trial runs on the demo unit. A broad user base is also expected to be built upon establishing the center, as there are numerous investigators with similar research interests at the University. The technology is proven and has been available since 2006; however, this would be the first such instrument to be installed on the campus as a core resource and made available to the entire Hopkins research community. High throughput functional characterization of the bioenergetic profile of cells is an unmet need that has lagged behind the information explosion generated by genomics, proteomics, and metabolomics. Thus, the only hope of understanding the role of an expanding number of disease-associated changes in proteins involved in energy metabolism will be to replace the tedious and time-consuming classical respiration or metabolite assays with multiplexed time-resolved recordings that are enabled by the Seahorse Bioscience instrument.