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使用微量移液器检测微粒的DNA涂层。

Sensing DNA-coatings of microparticles using micropipettes.

作者信息

Steinbock L J, Stober G, Keyser U F

机构信息

Institute for Experimental Physics I, University of Leipzig, Linné Str. 5, 04103 Leipzig, Germany.

出版信息

Biosens Bioelectron. 2009 Apr 15;24(8):2423-7. doi: 10.1016/j.bios.2008.12.026. Epub 2008 Dec 25.

Abstract

The resistive pulse technique is widely used to detect the size of small particles in aqueous solutions. This work demonstrates that a few tens of DNA molecules and thus the charges on a particle can be simply detected by pressure-driven translocation through a microcapillary based Coulter counter. The typical opening of the capillaries ranges from 2 to 6 microm. The custom-built system gives optical access using a high numerical aperture objective allowing to observe colloids passing the sensing volume by optical means. We show the feasibility of our setup by distinguishing colloids with one and two micron diameters. Our measurements prove that a few ten strands of DNA bound to the colloids can be detected. This can be achieved by simple comparison of current amplitudes for blank and coated colloids at low salt concentrations (2-40 mmol [NaCl]). Our results clearly demonstrate that the Coulter counter can be used to detect the surface charges on colloids. Moreover, the results are in good agreement with a dynamical computer model taking into account the full geometry of the capillary.

摘要

电阻脉冲技术被广泛用于检测水溶液中小颗粒的大小。这项工作表明,通过基于微毛细管的库尔特计数器进行压力驱动的转运,可以简单地检测几十条DNA分子,从而检测颗粒上的电荷。毛细管的典型开口范围为2至6微米。定制系统使用高数值孔径物镜提供光学通道,从而能够通过光学手段观察胶体通过传感体积。我们通过区分直径为1微米和2微米的胶体来展示我们装置的可行性。我们的测量证明,可以检测到与胶体结合的几十条DNA链。这可以通过在低盐浓度(2 - 40 mmol [NaCl])下简单比较空白胶体和包覆胶体的电流幅度来实现。我们的结果清楚地表明,库尔特计数器可用于检测胶体上的表面电荷。此外,结果与考虑毛细管完整几何形状的动态计算机模型非常吻合。

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