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）：零模波导（ZMG）是一种用于减少荧光光学检测系统中被照射流体体积的强大的新技术。该方法已被证明可以实现前所未有的小观察体积，从而带来检测和表征单分子的新能力。照明体积的减少可显着提高信噪比、信号背景比和时间分辨率。 Nanofluidics, Inc. 打算将这些结构商业化提供给工业和学术用户，以实现单分子检测的研究和应用。一项具有巨大商业潜力的应用已经被确定——一种需要信噪比的单分子 DNA 测序策略，而目前只能通过 ZMG 技术来实现。当研究界能够访问 ZMG 时，预计会出现其他应用程序。 ZMG的性能已经得到证实，因此商业化的剩余技术风险参数是工艺一致性和生产成本。第一阶段计划的目标是解决与 ZMG 生产一致性相关的问题。首先，必须开发一种工具来测量 ZMG 的内部尺寸，这是传统检测技术无法实现的。我们建议通过光学测量与模拟的比较，从 ZMG 的光学特性推断结构的尺寸。我们将使用有限元方法来模拟 ZMG 内部和周围的电磁场，以将各种几何形状的荧光激发曲线和偶极子辐射逃逸效率制成表格。然后，将使用一维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来制造和测试 ZMG，从而验证模型并测量制造变异性。成功将被定义为证明能够一致实现低于 1 阿托升的有效观测体积，且直径变异性小于 25%。完成第一阶段目标后，Nanfluidics, Inc. 将提交 SBIR 第二阶段提案，根据该提案，我们将使用现有方法制造设备并将其放置在 beta 测试站点，同时探索降低制造成本的方法。这项探索的范围包括从第一阶段验证的工艺的渐进改进到不需要昂贵的直写电子束光刻的新颖制造技术。