Characterization of optically-driven microstructures for manipulating single DNA molecules under a fluorescence microscope
Characterization of optically-driven microstructures for manipulating single DNA molecules under a fluorescence microscope
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荧光显微镜下操纵单个 DNA 分子的光学驱动微观结构的表征
DOI：
10.1049/iet-nbt.2015.0036
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发表时间：
2015
影响因子：
2.3
通讯作者：
F. Oohira
中科院分区：
文献类型：
--
作者：
K. Terao*;C. Masuda;R. Inukai;M. Gel;H. Oana;M. Washizu;T. Suzuki;H. Takao;F. Shimokawa;F. Oohira
光学镊子是使用荧光显微镜操纵单个DNA分子的强大工具，特别是在基于纳米技术的DNA分析中。我们之前提出了一种使用光学镊子驱动的微结构的操作技术，该技术允许处理传统技术无法操作的毫米长度的单个巨大DNA分子。为了进一步发展这一技术，作者在荧光显微镜下从制造和DNA操作效率的角度定量地表征了微观结构。并对其制作的成功率和精度进行了评价。结果表明，在浓度为106个粒子/ml的水溶液中，在精度为~ 50 nm的条件下获得了微观结构。这些微结构在荧光显微镜下的可见性也进行了表征，并阐明了微调可见性所需的制造参数。用微观结构操纵酵母染色体DNA分子说明了操作效率与微观结构几何形状之间的关系。该报告为基于现场DNA操作的单DNA分子分析中使用的微结构设计提供了指导，并有望在未来扩大该技术的应用。
Optical tweezers are powerful tools for manipulating single DNA molecules using fluorescence microscopy, particularly in nanotechnology‐based DNA analysis. We previously proposed a manipulation technique using microstructures driven by optical tweezers that allows the handling of single giant DNA molecules of millimetre length that cannot be manipulated by conventional techniques. To further develop this technique, the authors characterised the microstructures quantitatively from the view point of fabrication and efficiency of DNA manipulation under a fluorescence microscope. The success rate and precision of the fabrications were evaluated. The results indicate that the microstructures are obtained in an aqueous solution with a precision ∼50 nm at concentrations in the order of 106particles/ml. The visibility of these microstructures under a fluorescence microscope was also characterised, along with the elucidation of the fabrication parameters needed to fine tune visibility. Manipulating yeast chromosomal DNA molecules with the microstructures illustrated the relationship between the efficiency of manipulation and the geometrical shape of the microstructure. This report provides the guidelines for designing microstructures used in single DNA molecule analysis based on on‐site DNA manipulation, and is expected to broaden the applications of this technique in the future.