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The design and construction of novel near-infrared time-correlated single photon counting devices for the identification of analytes in multiplexed applications.

机译：新型近红外时间相关的单光子计数装置的设计和构造，用于在多重应用中鉴定分析物。

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摘要

This manuscript details the design, construction, and application of novel near infrared time correlated single photon counting devices to the identification of analytes in analytical separations. The thrust of this research is to provide a simple, low cost technique for the high-speed identification of DNA sequencing bases that are labeled with a series of unique near infrared fluorophores. These fluorophores are unique because they possess the same emission and absorption maxima, but different fluorescence lifetimes. Consequently, they allow analytes to be discriminated by fluorescence lifetime as opposed to color.; The first goal of this dissertation research was to implement a time correlated single photon counting system with the use of single mode fiber optics. Utilizing a passively mode locked Ti: Sapphire Laser, a single photon avalanche diode, single mode fiber optics and a mechanical switch a fiber optic based time correlated single photon counting device with subnanosecond resolution was constructed. The experimental results showed that group velocity dispersion was low and that it was possible to perform multiple time correlated single photon counting experiments with a limited number of excitation sources and detectors. It was determined that the average instrumental response of each channel was 181 picoseconds. The fluorescence lifetime of a near infrared dye, aluminum tetrasulfonated naphthalocyanine was determined to be 3.08 nanoseconds.; The second phase of this doctoral research involved the construction and characterization of a near infrared time correlated single photon counting scanning device. This integrated device consisted of a pulsed diode laser, single photon avalanche diode, and a time correlated single photon counting board. The instrument response function of this system was determined to be less than 300 ps. The sensitivity and ability to discriminate between various fluorophores was determined. In addition to its application for scanning solid surfaces such as DNA microarrays, the device was utilized to detect analytes in a micro-capillary electrophoresis separation. The fluorescence lifetimes of these analytes were determined on-line.
机译：该手稿详细介绍了新型近红外时间相关的单光子计数设备的设计，构造和应用，以鉴定分析分离中的分析物。这项研究的目的是提供一种简单，低成本的技术，用于高速鉴定DNA测序碱基，该碱基被一系列独特的近红外荧光团标记。这些荧光团是独特的，因为它们具有相同的发射和吸收最大值，但荧光寿命不同。因此，它们可以通过荧光寿命而不是颜色来区分分析物。本论文研究的首要目标是利用单模光纤实现与时间相关的单光子计数系统。利用被动模式锁定的钛蓝宝石激光器，单光子雪崩二极管，单模光纤和机械开关，构建了基于光纤的时间相关的单光子计数装置，其纳秒分辨率。实验结果表明，群速度色散较低，并且可以使用有限数量的激发源和检测器进行多次与时间相关的单光子计数实验。确定每个通道的平均仪器响应为181皮秒。经测定，近红外染料四磺化萘酞菁铝的荧光寿命为3.08纳秒。该博士研究的第二阶段涉及近红外时间相关的单光子计数扫描设备的构造和表征。该集成设备由脉冲二极管激光器，单光子雪崩二极管和与时间相关的单光子计数板组成。确定该系统的仪器响应功能小于300 ps。确定了区分各种荧光团的灵敏度和能力。除了用于扫描固体表面（例如DNA微阵列）外，该设备还用于检测微毛细管电泳分离中的分析物。在线测定这些分析物的荧光寿命。

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  • 作者

    Waddell, Emanuel Austin, Jr.;

  • 作者单位

    Louisiana State University and Agricultural & Mechanical College.;

  • 授予单位 Louisiana State University and Agricultural & Mechanical College.;
  • 学科 Chemistry Analytical.
  • 学位 Ph.D.
  • 年度 2000
  • 页码 140 p.
  • 总页数 140
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 化学;
  • 关键词

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