LaserLaB Amsterdam and Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, Netherlands.
Methods Mol Biol. 2024;2694:403-420. doi: 10.1007/978-1-0716-3377-9_19.
Magnetic tweezers are a force spectroscopy single-molecule technique. They enable the mechanical manipulation of biomolecules via the means of a magnetic particle under an attractive force applied by a magnetic field source. The magnetic particle is tethered to the glass surface of a flow chamber by the biomolecule, and functionalization strategies have been developed to reduce the nonspecific interactions of either the magnetic particles or biomolecules with the surface. Here, we describe two complementary strategies to achieve a high tether density while reducing the interactions of both the magnetic particle and the biomolecule of interest with the glass surface. Using a large detector CMOS camera, the simultaneous observation of several hundreds of tethered magnetic beads is achievable, allowing high-throughput single-molecule measurements. We further describe here a simple procedure to perform the calibration in force of a magnetic tweezers assay.
磁镊是一种力谱单分子技术。它们通过磁场源施加的吸引力，使磁珠对生物分子进行机械操作。磁珠通过生物分子与流动室的玻璃表面连接，已经开发出功能化策略来减少磁珠或生物分子与表面的非特异性相互作用。在这里，我们描述了两种互补的策略，以在降低磁珠和感兴趣的生物分子与玻璃表面相互作用的同时，实现高连接密度。使用大探测器 CMOS 相机，可以同时观察几百个连接的磁珠，从而实现高通量单分子测量。我们还在这里描述了一种简单的程序来执行磁镊测定的力校准。
