Zero-Mode Waveguides for Single-Molecule Detection

用于单分子检测的零模式波导

• 批准号: 6693874
• 负责人: STEPHEN WHITFIELD TURNER
• 金额: $ 11.03万
• 依托单位: PACIFIC BIOSCIENCES OF CALIFORNIA, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6693874
• 关键词: bioimaging /biomedical imaging; biomedical equipment development; electric field; electron optics; fluorescence microscopy; fluorescence spectrometry; high throughput technology; method development; nanotechnology; optics

DESCRIPTION (provided by applicant): Zero-mode waveguides (ZMGs) are a new and powerful technique for reducing the volume of illuminated fluid in a fluorescence optical detection system. The method has been shown to allow unprecedented small observation volumes, resulting in new abilities in detecting and characterizing single molecules. Reduction of illuminated volume provides significant gains in signal-to-noise, signal-to-background and temporal resolution. Nanofluidics, Inc. intends to make these structures commercially available to industrial and academic users to enable research in and applications of single-molecule detection. One application with significant commercial potential has already been identified-- a single-molecule DNA sequencing strategy that requires a signal-to-noise that right now can only be attained with the ZMG technology. Other applications are expected to emerge when the research community is provided access the ZMGs. The performance of the ZMG has been proven, so the remaining technical risk parameters to commercialization are process consistency and production cost. The goals of this phase I program are to address the issues related to consistency in the production of ZMGs. First, it is essential to develop a tool to measure the interior dimensions of the ZMG, which is not possible with conventional inspection techniques. We propose to infer the dimensions of the structure from optical characteristics of the ZMG by comparison of optical measurements with simulation. We will use finite-element methods to simulate the electromagnetic fields in and around the ZMGs to tabulate both the fluorescence excitation profile and the efficiency of dipole radiation escape for a variety of geometries. Then a one-dimensional analytic diffusion model will be used to predict the fluorescence correlation spectra expected for each. Next, ZMGs will be fabricated and tested with the same geometries allowing validation of the models and measurement of fabrication variability. Success will be defined as demonstration of consistent attainability of sub 1-attoliter effective observation volumes, with variability in diameters of less than 25%. Upon completion of the phase I goals, Nanofluidics, Inc. will submit an SBIR phase II proposal under which we will make devices using the present methods and place them in beta test sites, while concurrently exploring methods of reducing the cost of manufacture. This exploration will range from incremental improvements of the process proven in Phase I to novel fabrication techniques that don't require expensive direct-write electron-beam lithography.

描述（由申请人提供）：零模式波导（ZMG）是一种新的功能强大技术，用于减少荧光光学检测系统中照明流体的体积。该方法已被证明可以允许前所未有的小观察体积，从而在检测和表征单分子方面具有新的能力。减少照明体积可在信噪，信噪比和时间分辨率方面显着增长。 Nanofluidics，Inc。打算让工业和学术用户在商业上使用这些结构，以实现对单分子检测的研究和应用。已经确定了一种具有巨大商业潜力的应用程序 - 一种单分子DNA测序策略，该策略需要一个信噪比，该策略只能使用ZMG技术才能实现。当提供研究社区访问ZMG时，预计其他应用程序将出现。 ZMG的性能已得到证明，因此商业化的剩余技术风险参数是过程一致性和生产成本。该阶段I计划的目标是解决与ZMG生产一致性有关的问题。首先，必须开发一种工具来测量ZMG的内部维度，而常规检查技术是不可能的。我们建议通过将光学测量与仿真进行比较，从ZMG的光学特征中推断出结构的尺寸。我们将使用有限元方法模拟ZMG中和周围的电磁场，以将荧光激发曲线和偶极辐射的效率同时制成各种几何形状。然后，将使用一维的分析扩散模型来预测每个预期的荧光相关光谱。接下来，将使用相同的几何形状制造和测试ZMG，允许验证模型并测量制造可变性。成功将定义为表明亚1近视有效观察量的一致可达到的能力，直径少于25％。 I阶段目标完成后，Nanofluidics，Inc。将提交SBIR II期提案，我们将使用当前方法制作设备并将其放置在Beta测试站点中，同时探索降低制造成本的方法。这种探索的范围从I阶段证明的工艺的增量改进到不需要昂贵的直接连接电子束光刻的新型制造技术。