New and Improved Sensor Platforms and Quantification of Nitric Oxide for In Vitro and In Vivo Systems

项目摘要/摘要 一氧化氮(NO)是生物系统中的一种关键信号分子，但由于缺乏对NO的了解，人们对其了解较少。 时间和空间传感器测量NO。一种直接、快速、准确地测量NO的传感器将是 对正在进行的和未来的研究具有巨大的价值。单壁碳纳米管(SWNT)具有优异的性能 在体外和体内都有可能用作NO传感器，但它们并没有被研究界广泛使用 因为它们在易于使用的平台中并不容易获得。这个项目的目标是1)开发新的 SWNT传感器平台和修改当前的系统，以提高传感器的专用性和处理能力，以及2) 用高灵敏度、高精度的传感器测定一氧化氮浓度和动态。目标将是 通过完成四个具体项目完成。在第一个项目中，我们将开发一个比率计量器 传感器没有经过长时间的量化，第二个项目将涉及开发一款简单易用的 用于体内传递和稳定能够同时具有空间和时间能力的SWNT传感器的平台 分析物定量。项目3和4将使用SWNT传感器来调查NO浓度，特别是 在项目3中，我们将确定细胞内NO与细胞器和细胞膜的动态关系，以及 项目4将涉及调查细胞外NO浓度的值和与 健康的和患病的细胞。 这项工作将利用艾弗森实验室的专业知识来设计新的和改进的传感器交付 平台，并使用传感器来研究细胞内和细胞外的NO信号。通过开发这些 平台还将提供用于开发其他SWNT传感器系统的模板，允许 研究人员将在空间和时间上了解活性氧物种、小分子和蛋白质 在亚细胞层面上的时尚。本项目中被调查细胞的NO信号的研究将 使研究人员对NO在细胞信号转导中的作用有一个基本的了解，并为 对其他细胞类型的研究。

PROJECT SUMMARY/ABSTRACT Nitric oxide (NO) is a key signaling molecule in biological systems, but it is poorly understood due to a lack of temporal and spatial sensors that measure NO. A direct, fast and accurate sensor for measuring NO would be of immense value to ongoing and future research. Single walled carbon nanotubes (SWNT) have excellent potential for use as NO sensors both in vitro and in vivo, but they are not widely used by the research community because they are not readily available in an easy-to-use platform. The goals of this project are to 1) develop new SWNT sensor platforms and modify the current systems to improve sensor specificity and handling, and 2) determine nitric oxide concentration and dynamics with a highly sensitive and accurate sensor. The goals will be accomplished through the completion of four specific projects. In the first project we will develop a ratiometric sensor to quantify NO over long time periods, the second project will involve the development of an easy-to-use platform for the in vivo delivery and stabilization of SWNT sensors that are capable of both spatial and temporal analyte quantification. Projects 3 and 4 will use the SWNT sensors to investigate NO concentrations, specifically for project 3 we will determine the intracellular dynamics of NO in relation to organelles and membranes and project 4 will involve the investigation of extracellular NO concentration values and gradients associated with healthy and diseased cells. This work will leverage the expertise of the Iverson Laboratory to engineer new and improved sensor delivery platforms and use the sensors to investigate intracellular and extracellular NO signaling. By developing these platforms a template for the development of other SWNT sensor systems will also be provided, allowing researchers to learn about reactive oxygen species, small molecules and proteins in a spatial and temporal fashion at the sub-cellular level. The investigation of the NO signaling for the cells investigated in this project will provide researchers with a basic understanding of NO’s role in cell signaling and provide a template for investigation of other cell types.