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Three-Dimensional Tracking of Tethered Particles for Probing Nanometer-Scale Single-Molecule Dynamics Using a Plasmonic Microscope
ACS Sensors ( IF 8.2 ) Pub Date : 2021-11-17 , DOI: 10.1021/acssensors.1c01927
Guangzhong Ma 1 , Zijian Wan 1, 2 , Yunze Yang 1 , Wenwen Jing 1, 3 , Shaopeng Wang 1, 4
Affiliation  

Three-dimensional (3D) tracking of surface-tethered single particles reveals the dynamics of the molecular tether. However, most 3D tracking techniques lack precision, especially in the axial direction, for measuring the dynamics of biomolecules with a spatial scale of several nanometers. Here, we present a plasmonic imaging technique that can track the motion of ∼100 tethered particles in 3D simultaneously with sub-nanometer axial precision and single-digit nanometer lateral precision at millisecond time resolution. By tracking the 3D coordinates of a tethered particle with high spatial resolution, we are able to determine the dynamics of single short DNA and study its interaction with enzymes. We further show that the particle motion pattern can be used to identify specific and nonspecific interactions in immunoassays. We anticipate that our 3D tracking technique can contribute to the understanding of molecular dynamics and interactions at the single-molecule level.

中文翻译:

使用等离子显微镜对系留粒子进行三维跟踪以探测纳米级单分子动力学

表面束缚的单个粒子的三维 (3D) 跟踪揭示了分子束缚的动力学。然而,大多数 3D 跟踪技术缺乏精度,尤其是在轴向方向上,无法测量具有几纳米空间尺度的生物分子的动力学。在这里,我们提出了一种等离子体成像技术,可以在 3D 中同时跟踪 ~100 个系留粒子的运动,具有亚纳米级轴向精度和单位数纳米横向精度,时间分辨率为毫秒。通过跟踪具有高空间分辨率的系留粒子的 3D 坐标,我们能够确定单个短 DNA 的动力学并研究其与酶的相互作用。我们进一步表明,粒子运动模式可用于识别免疫测定中的特异性和非特异性相互作用。
更新日期:2021-11-26
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