IDBR: Development of an Optical Sensor for Biological S-Nitrosothiols

IDBR：生物 S-亚硝基硫醇光学传感器的开发

• 批准号: 0964178
• 负责人: Kevin Lehmann
• 金额: $ 15万
• 依托单位: University of Virginia Main Campus
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0964178&HistoricalAwards=false
• 关键词: IDBR; Development; Optical; Sensor; Biological

A new mechanism for cellular communication, S-nitrosylation, has recently been discovered. This mechanism involves changes in proteins caused by chemical reactions between sulfur atoms and a compound known as nitrosonium; the products of these reactions are known as S-nitrosothiols. These chemical groups have been demonstrated to be important post-translational protein modifications and are part of many signaling pathways in living organisms. However, research in this area has been hampered by the lack of instrumentation that can measure the level of S-nitrosothiols with sufficient sensitivity and speed for many cases of interest to biologists. New instrumentation, based upon continuous wave, cavity ring-down spectroscopy (CRDS) is being developed with a goal to detect Nitric Oxide (NO) in an inert gas at the single digit parts per trillion level. NO can be liberated from tissue either chemically or photochemically and the resulting gas flushed out into analyzer. The instrument measure the optical loss is an optical cavity made from mirrors with 99.97% reflectivity, which will result in an effective absorption pathlength of over 3000 passes through the sample. A recently introduced laser, external cavity, quantum cascade laser (ec-QCL) will be used to efficiently excite the optical resonances of the cavity and will be tuned across the rotational transitions in the strong stretching fundamental transition of the NO molecule near 5.2 micro-meter. The concentration of NO is determined by an increase in the decay rate of intensity of light in the cavity when the laser wavelength is tuned to a NO absorption line. Based upon the established sensitivity of similar CRDS instruments working in the near-InfraRed, a lower detection limit of approximately three orders of magnitude higher than for the existing technology is predicted, which is based upon chemiluminescence. Further, this instrument should give dramatically improved temporal resolution, require reduced sample sizes, and will distinguish isotopic forms of NO, allowing labeling experiments to be done to determine the chemical origin of the released NO. The first year of the project will be devoted towards design, construction, and testing of the CRDS based instrument. The second will be devoted towards interfacing the CRDS instrument with the biological applications. In addition to applications in biological research, the instrumentation could potentially be used for medical diagnostics. Progress on this project will be reported at http://faculty.virginia.edu/lehmannlab/NO_Detection.html.

最近发现了一种新的细胞通讯机制——s -亚硝基化。这种机制涉及到由硫原子和一种被称为亚硝基铵的化合物之间的化学反应引起的蛋白质变化；这些反应的产物被称为s -亚硝基硫醇。这些化学基团已被证明是重要的翻译后蛋白质修饰，是生物体中许多信号通路的一部分。然而，这一领域的研究一直受到阻碍，因为缺乏能够以足够的灵敏度和速度测量生物学家感兴趣的许多病例的s -亚硝基硫醇水平的仪器。基于连续波腔衰荡光谱（CRDS）的新仪器正在开发中，其目标是检测惰性气体中万亿分之一的一氧化氮（NO）。NO可以通过化学或光化学方法从组织中释放出来，产生的气体被冲洗到分析仪中。仪器测量的光损耗是由反射率为99.97%的镜面制成的光腔，这将导致有效吸收光路长度超过3000次通过样品。最近推出的激光，外腔，量子级联激光器（ec-QCL）将用于有效地激发腔的光学共振，并将在5.2微米附近的NO分子的强拉伸基本跃迁中进行旋转跃迁调谐。当激光波长调谐到NO吸收线时，由腔内光强度衰减率的增加决定了NO的浓度。基于在近红外工作的类似CRDS仪器的灵敏度，预测了一个比现有技术高约三个数量级的低检测极限，这是基于化学发光的。此外，该仪器将大大提高时间分辨率，减少样本量，并将区分NO的同位素形式，允许进行标记实验，以确定释放的NO的化学来源。项目的第一年将致力于基于CRDS的仪器的设计、建造和测试。第二个将致力于将CRDS仪器与生物应用相结合。除了在生物学研究中的应用外，该仪器还可能用于医学诊断。该项目的进展情况将在http://faculty.virginia.edu/lehmannlab/NO_Detection.html上报告。