Simultaneous, coincident optical trapping and single-molecule fluorescence

被引：0
作者：
Lang M.J. [1 ,2 ,4 ,5 ,6 ]
Fordyce P.M. [1 ,3 ,4 ,6 ]
Fordyce A.M. [4 ]
Neuman K.C. [1 ,2 ,4 ,6 ]
Block S.M. [1 ,2 ,4 ,6 ]
机构：
[1] Department of Biological Sciences, Stanford University, Stanford, CA
[2] Department of Applied Physics, Stanford University, Stanford, CA
[3] Department of Physics, Stanford University, Stanford, CA
[4] Stanford University, Stanford, CA
[5] Biological Engineering and Department of Mechanical Engineering, Institute of Technology, Cambridge, MA, 02139-4307
[6] Department of Biological Sciences, Stanford University, Stanford, CA
来源：
Nature Methods | 2004年 / 1卷 / 2期
基金：
美国国家科学基金会; 美国国家卫生研究院;
关键词：
D O I：
10.1038/nmeth714
中图分类号：
学科分类号：
摘要：
We constructed a microscope-based instrument capable of simultaneous, spatially coincident optical trapping and single-molecule fluorescence. The capabilities of this apparatus were demonstrated by studying the force-induced strand separation of a dye-labeled, 15-base-pair region of double-stranded DNA (dsDNA), with force applied either parallel (‘unzipping’ mode) or perpendicular (‘shearing’ mode) to the long axis of the region. Mechanical transitions corresponding to DNA hybrid rupture occurred simultaneously with discontinuous changes in the fluorescence emission. The rupture force was strongly dependent on the direction of applied force, indicating the existence of distinct unbinding pathways for the two force-loading modes. From the rupture force histograms, we determined the distance to the thermodynamic transition state and the thermal off rates in the absence of load for both processes. © 2004 Nature Publishing Group.
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页码：133 / 139
页数：6
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