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

项目总结/摘要 一氧化氮（NO）是生物系统中的关键信号分子，但由于缺乏对NO的研究，对其了解甚少。 用于测量NO的直接、快速和精确的传感器将 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 作为NO传感器在体外和体内都有潜力，但它们没有被研究界广泛使用 因为它们在易于使用的平台中不容易获得。该项目的目标是：1）开发新的 SWNT传感器平台，并修改当前系统以提高传感器特异性和处理，以及2） 使用高灵敏度和精确的传感器确定一氧化氮浓度和动力学。目标将是 通过四个具体项目的完成。在第一个项目中，我们将开发一个比率计量 传感器，以量化NO在很长一段时间内，第二个项目将涉及一个易于使用的开发 用于SWNT传感器的体内递送和稳定化的平台，所述SWNT传感器能够空间和时间两个方面的功能。 分析物定量。项目3和项目4将使用SWNT传感器来研究NO浓度， 对于项目3，我们将确定NO与细胞器和膜的关系， 项目4将涉及研究细胞外NO浓度值和梯度与 健康和患病的细胞。 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NO信号。通过开发这些 还将提供用于开发其他SWNT传感器系统的平台模板， 研究人员了解活性氧，小分子和蛋白质在一个空间和时间 在亚细胞水平的时尚。本项目中研究的细胞的NO信号传导的研究将 为研究人员提供了NO在细胞信号传导中的作用的基本理解，并为 研究其他细胞类型。

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.