title: Characterisation of optically driven microstructures for manipulating single DNA molecules under a fluorescence microscope.
abstract: 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 10(6) particles/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.
authors.affiliation: JST-PREST, Saitama 332-0012, Japan. terao@eng.kagawa-u.ac.jp.
authors.forename: Kyohei
authors.initials: K
authors.lastname: Terao
authors.affiliation: Department of Intelligent Mechanical Systems Engineering, Kagawa University, Takamatsu 761-0396, Japan.
authors.forename: Chihiro
authors.initials: C
authors.lastname: Masuda
authors.affiliation: Department of Intelligent Mechanical Systems Engineering, Kagawa University, Takamatsu 761-0396, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Inukai
authors.affiliation: CSIRO, Material Science and Engineering, Clayton, VIC 3030, Australia.
authors.forename: Murat
authors.initials: M
authors.lastname: Gel
authors.affiliation: Department of Mechanical Engineering, The University of Tokyo, Tokyo 113-8656, Japan.
authors.forename: Hidehiro
authors.initials: H
authors.lastname: Oana
authors.affiliation: Department of Mechanical Engineering, The University of Tokyo, Tokyo 113-8656, Japan.
authors.forename: Masao
authors.initials: M
authors.lastname: Washizu
authors.affiliation: Department of Intelligent Mechanical Systems Engineering, Kagawa University, Takamatsu 761-0396, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Suzuki
authors.affiliation: Department of Intelligent Mechanical Systems Engineering, Kagawa University, Takamatsu 761-0396, Japan.
authors.forename: Hidekuni
authors.initials: H
authors.lastname: Takao
authors.affiliation: Department of Intelligent Mechanical Systems Engineering, Kagawa University, Takamatsu 761-0396, Japan.
authors.forename: Fusao
authors.initials: F
authors.lastname: Shimokawa
authors.affiliation: Department of Intelligent Mechanical Systems Engineering, Kagawa University, Takamatsu 761-0396, Japan.
authors.forename: Fumikazu
authors.initials: F
authors.lastname: Oohira
mesh_terms: D004247:DNA / Q000737:chemistry*; D004867:Equipment Design; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D055616:Microtechnology / Q000295:instrumentation*; D052898:Optical Tweezers
chemical_list: D004247:DNA