DNA conformation on surfaces measured by fluorescence self-interference
DNA conformation on surfaces measured by fluorescence self-interference
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DOI：
10.1073/pnas.0511214103
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发表时间：
2006-02-21
影响因子：
11.1
通讯作者：
Cantor, CR
中科院分区：
文献类型：
--
作者：
Moiseev, L;Ünlü, MS;Cantor, CR
附着在微阵列表面的 DNA 分子的构象可能会显着影响杂交的效率。尽管已经应用了多种方法来确定 DNA 层的结构，但它们对 DNA 分子形状的变化不是很敏感。在这里，我们描述了一种称为光谱自干涉荧光显微镜的干涉技术的应用，以精确测量 DNA 层中荧光标记相对于表面的平均位置，从而确定表面结合 DNA 分子构象的具体信息。利用光谱自干涉荧光显微镜，我们估计了卷曲单链DNA的形状、不同长度双链DNA的平均倾斜度以及杂交量。这些数据为表面 DNA 分子分布的新型光学表面分析方法的能力提供了重要的概念证明。表面 DNA 构象和杂交行为的测定提供了推动 DNA 界面应用向前发展所需的信息，从而影响新兴的临床和生物技术领域。
The conformation of DNA molecules tethered to the surface of a microarray may significantly affect the efficiency of hybridization. Although a number of methods have been applied to determine the structure of the DNA layer, they are not very sensitive to variations in the shape of DNA molecules. Here we describe the application of an interferometric technique called spectral self-interference fluorescence microscopy to the precise measurement of the average location of a fluorescent label in a DNA layer relative to the surface and thus determine specific information on the conformation of the surf ace-bound DNA molecules. Using spectral self-interference fluorescence microscopy, we have estimated the shape of coiled single-stranded DNA, the average tilt of double-stranded DNA of different lengths, and the amount of hybridization. The data provide important proofs of concept for the capabilities of novel optical surface analytical methods of the molecular disposition of DNA on surfaces. The determination of DNA conformations on surfaces and hybridization behavior provide information required to move DNA interfacial applications forward and thus impact emerging clinical and biotechnological fields.