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用于测量扭转和扭矩的磁性镊子。

Magnetic tweezers for the measurement of twist and torque.

作者信息

Lipfert Jan, Lee Mina, Ordu Orkide, Kerssemakers Jacob W J, Dekker Nynke H

机构信息

Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology.

Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology;

出版信息

J Vis Exp. 2014 May 19(87):51503. doi: 10.3791/51503.

Abstract

Single-molecule techniques make it possible to investigate the behavior of individual biological molecules in solution in real time. These techniques include so-called force spectroscopy approaches such as atomic force microscopy, optical tweezers, flow stretching, and magnetic tweezers. Amongst these approaches, magnetic tweezers have distinguished themselves by their ability to apply torque while maintaining a constant stretching force. Here, it is illustrated how such a "conventional" magnetic tweezers experimental configuration can, through a straightforward modification of its field configuration to minimize the magnitude of the transverse field, be adapted to measure the degree of twist in a biological molecule. The resulting configuration is termed the freely-orbiting magnetic tweezers. Additionally, it is shown how further modification of the field configuration can yield a transverse field with a magnitude intermediate between that of the "conventional" magnetic tweezers and the freely-orbiting magnetic tweezers, which makes it possible to directly measure the torque stored in a biological molecule. This configuration is termed the magnetic torque tweezers. The accompanying video explains in detail how the conversion of conventional magnetic tweezers into freely-orbiting magnetic tweezers and magnetic torque tweezers can be accomplished, and demonstrates the use of these techniques. These adaptations maintain all the strengths of conventional magnetic tweezers while greatly expanding the versatility of this powerful instrument.

摘要

单分子技术使实时研究溶液中单个生物分子的行为成为可能。这些技术包括所谓的力谱方法,如原子力显微镜、光镊、流动拉伸和磁镊。在这些方法中,磁镊因其在保持恒定拉伸力的同时施加扭矩的能力而脱颖而出。在此,说明了如何通过对其场配置进行直接修改以最小化横向场的大小,将这种“传统”磁镊实验配置调整为测量生物分子中的扭转程度。由此产生的配置被称为自由轨道磁镊。此外,还展示了如何进一步修改场配置以产生一个横向场,其大小介于“传统”磁镊和自由轨道磁镊之间,这使得直接测量生物分子中存储的扭矩成为可能。这种配置被称为磁扭矩镊。随附的视频详细解释了如何将传统磁镊转换为自由轨道磁镊和磁扭矩镊,并展示了这些技术的使用。这些改进保留了传统磁镊的所有优点,同时极大地扩展了这种强大仪器的多功能性。

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