Zero-mode waveguides for single-molecule analysis at high concentrations
Zero-mode waveguides for single-molecule analysis at high concentrations
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DOI：
10.1126/science.1079700
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发表时间：
2003-01-31
期刊：
影响因子：
56.9
通讯作者：
Webb, WW
中科院分区：
文献类型：
--
作者：
Levene, MJ;Korlach, J;Webb, WW
用于观察单个分子的动力学的光学方法需要皮科至纳摩尔浓度的荧光团以分离单个分子。然而，许多生物相关的过程发生在微摩尔配体浓度，需要减少在传统的观察体积的三个数量级。我们表明，阵列的零模波导组成的亚波长孔在金属膜提供了一个简单的和高度平行的手段，研究单分子动力学在微摩尔浓度与微秒的时间分辨率。我们目前的观察DNA聚合酶的活性作为一个例子的有效性的零模波导进行单分子实验在高浓度。
Optical approaches for observing the dynamics of single molecules have required pico- to nanomolar concentrations of fluorophore in order to isolate individual molecules. However, many biologically relevant processes occur at micromolar ligand concentrations, necessitating a reduction in the conventional observation volume by three orders of magnitude. We show that arrays of zero-mode waveguides consisting of subwavelength holes in a metal film provide a simple and highly parallel means for studying single-molecule dynamics at micromolar concentrations with microsecond temporal resolution. We present observations of DNA polymerase activity as an example of the effectiveness of zero-mode waveguides for performing single-molecule experiments at high concentrations.