A novel electromagnetic apparatus for rapid multiplex single molecule force spectroscopy.
A novel electromagnetic apparatus for rapid multiplex single molecule force spectroscopy.
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DOI：
10.1166/jnn.2013.6099
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发表时间：
2013-02
影响因子：
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通讯作者：
Yi Shen;D. Czajkowsky;Xiaowei Li;Jielin Sun;Jun Hu;Zhifeng Shao
中科院分区：
文献类型：
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作者：
Yi Shen;D. Czajkowsky;Xiaowei Li;Jielin Sun;Jun Hu;Zhifeng Shao
单分子力光谱学已经彻底改变了我们探测分子结构和相互作用细节的能力，但是为了获得统计上可靠的结果，所需的单个测量的数量有时会令人生畏。为了克服这个问题，最近开发了许多能够同时监测数十个单独生物分子行为的仪器。在这项工作中，我们构建了一种新型的电磁装置，利用磁性微球进行多路单分子力测量。在该系统中，磁场是由电子显微镜的电子透镜与高压闪光灯电路配合产生的，以快速获得稳定的磁场。我们表明，该仪器可以产生一个均匀的磁力高达-20 pN在5 ms内，在一个区域跨越1 mm。该装置的成功应用，以力依赖性的延伸dsDNA充分验证了这种方法。此外，极片的透镜状设计与光学成像完全兼容，从而允许整合单分子荧光能力，这将使该系统成为用于相互作用的单分子内的快速过程的多维表征的特别强大的装置。
Single-molecule force spectroscopy has revolutionized our ability to probe the details of molecular structures and interactions, but the numbers of individual measurements required for achieving a statistically reliable result can sometimes prove daunting. To overcome this problem, a number of instruments have recently been developed that are capable of monitoring the behavior of tens of individual biomolecules simultaneously. In this work, we have constructed a novel electromagnetic apparatus for multiplex single molecule force measurements utilizing magnetic microspheres. In this system, the magnetic field is generated with an electron-lens of an electron microscope mated with a high voltage flash light circuit to rapidly attain a stable magnetic field. We show that this instrument can generate a uniform magnetic force of up to -20 pN within 5 ms, over a region spanning 1 mm. The successful application of this apparatus to the force-dependent extension of dsDNA fully validates this approach. Furthermore, the lens-like design of the pole piece is fully compatible with optical imaging, thus allowing for the integration of single molecule fluorescence capabilities that should make this system a particularly powerful apparatus for multi-dimensional characterization of fast processes within interacting single molecules.