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首页> 外文会议>NATO Advanced Research Workshop on Biological, Biophysical amp; Theoretical Aspects of Polymer Structure and Transport Jun 20-25, 1999 Bikal, Hungary >SINGLE MOLECULE NUCLEIC ACID ANALYSIS BY FLUORESCENCE FLOW CYTOMETRY
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SINGLE MOLECULE NUCLEIC ACID ANALYSIS BY FLUORESCENCE FLOW CYTOMETRY

机译：荧光流式细胞术分析单分子核酸

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摘要

During the past decade the sensitivity and selectivity of laser-induced fluorescence detection methods in liquids have been refined to the point where a molecule labeled with a single fluorophore dissolved in solution can be detected and identified as it flows through a focused excitation laser beam. Progress in this field is reviewed in a number of recent papers. Single molecule spectroscopy (SMS), the detection and measurement of properties of individual fluorescent molecules, allows assays to be done that would be difficult or impossible with bulk measurement methods. A useful result of SMS is the measurement of property distribution functions of single fluorescent molecules. Property distribution functions are accessible with SMS since the fluorescence properties of single molecules are measured individually. This information is often inaccessible to bulk measurements due to ensemble averaging. Photophysical properties that have been used for identification of fluorophores at the single molecule level include: gross differences in excitation and emission spectra, fluorescence emission lifetime, fluorescence emission intensity, and fluorescence emission polarization anisotropy. Two SMS assays under development in our laboratory, rapid sizing of single DNA fragments and the possibility of single molecule DNA sequencing, are discussed in sections 3 and 4 of this chapter.
机译：在过去的十年中，激光诱导的荧光检测方法在液体中的灵敏度和选择性已经提高到可以溶解并标记有溶解在溶液中的单个荧光团标记的分子流过聚焦激发激光束的程度。最近的一些论文对这一领域的进展进行了综述。单分子光谱法（SMS）是对单个荧光分子特性的检测和测量，它允许进行批量测量方法难以或不可能进行的测定。 SMS的有用结果是单个荧光分子特性分布函数的测量。 SMS可以访问属性分布函数，因为单个分子的荧光属性是单独测量的。由于集成平均，通常无法进行批量测量。用于鉴定单分子水平荧光团的光物理性质包括：激发和发射光谱的总体差异，荧光发射寿命，荧光发射强度和荧光发射偏振各向异性。本章第3和4节讨论了我们实验室中正在开发的两种SMS检测方法，即快速确定单个DNA片段的大小以及单分子DNA测序的可能性。

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