PROGRESS W/ SELF REFERENCING POLAROGRAPHIC ELECTRODES

自参考极谱电极的进展

• 批准号: 6568418
• 负责人: D M PORTERFIELD
• 金额: $ 21.13万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6568418
• 关键词: animal tissue; bioengineering /biomedical engineering; biomedical equipment development; biomedical resource; human tissue; technology /technique

Upon completion of the prototype self-referencing polarographic (Serp) system for the detection of cellular oxygen flux, the groups' effort has shifted to testing and refinement. There has also been considerable work done on understanding the analysis of the resulting data. Two types of testing have been completed using either artificial or biological sources of oxygen concentration gradients. Within the oxygen gradient the difference in oxygen concentration between two points can be used to determine oxygen flux based on Fick's law. While this is straight forward, understanding the steady-state dynamics of oxygen concentration within a gradient around a glass micropipette acting as a source oxygen, has proved more difficult. The results are that we better understand this system and can now apply this knowledge to other Serp-electrode techniques. The results of the testing of the Serp-oxygen electrodes in this artificial environment have shown that the system reliably, and accurately, measures oxygen flux. Furthermore, the sensitivity of the system has been much improved by a recent upgrade of the main amplifier. This improved instrumentation is good enough to allow spatial resolution of oxygen flux associated with changes in mitochondrial localization within a single cell. We have been using this new technology to measure oxygen flux around a variety of different types of single cells. The addition of temperature control capabilities to the Serp system has allowed us to measure oxygen flux in a variety of mammalian cells at normal physiological temperatures. Specifically, studies have been completed that examined metabolic oxygen consumption by mouse oocytes and developing embryos. These studies have allowed us to demonstrate that the electrodes are robust enough, despite their delicate nature, to operate at temperatures as high as 37o C. This is of paramount importance if the system is to be reliably applied to biomedical research. Clearly this technique has passed through the prototype stage and is now a significant resource to investigators at the Center. We are already witnessing interest and applications to use this technology from biomedical researchers. Given that the system and supporting electronics have been perfected, we can now shift our focus to the development of new microelectrode construction techniques that will allow us to build improved oxygen microelectrodes, as well as beginning the development of the Serp-nitric oxide detection system.

在完成原型后，自参考极谱 (SERP)细胞氧通量检测系统，小组的 努力已转向测试和改进。也有过 在理解对结果的分析方面所做的大量工作 数据。已经完成了两种类型的测试，使用的是 人工或生物源的氧气浓度梯度。 在氧气梯度内，氧气浓度的差异 两点之间的距离可以用来确定氧通量 菲克定律。虽然这是直接的，但理解 周围梯度内氧浓度的稳态动力学 一种玻璃微吸管作为氧气来源，已经证明了 很难。结果是我们更好地了解了这个系统，并 现在可以将这一知识应用于其他蛇行电极技术。这个 本院血氧电极的检测结果 人工环境已经表明，该系统可靠，并且 准确地测量氧气流量。此外，这一事件的敏感性 通过最近对主要设备的升级，系统得到了很大的改进 放大器。这种改进的工具足够好，可以允许 氧通量与大气环境变化相关的空间分辨率 线粒体在单个细胞内的定位。我们一直在使用 这项新技术可以测量各种气体周围的氧通量 不同类型的单细胞。增加了温度控制功能 SERP系统的能力使我们能够测量氧通量 在正常生理温度下的各种哺乳动物细胞中。 具体地说，已经完成了检查新陈代谢的研究 小鼠卵母细胞和发育中胚胎的耗氧量。这些 研究已经使我们能够证明电极是坚固的 尽管它们具有微妙的性质，但足以在 高达37摄氏度，这是至关重要的如果系统要 可靠地应用于生物医学研究。显然，这项技术具有 经历了原型阶段，现在是一个重要的资源 给中心的调查人员。我们已经见证了人们的兴趣 以及来自生物医学研究人员的使用这项技术的应用。 鉴于系统和配套电子设备已经完善， 我们现在可以把重点转移到开发新的微电极上了 建造技术将使我们能够建造更好的氧气 微电极，以及开始发展的微电极 SERP-一氧化氮检测系统。