Department of Engineering, University of Cambridge, Cambridge, United Kingdom.
Department of Engineering, University of Cambridge, Cambridge, United Kingdom.
Biophys J. 2018 Jul 3;115(1):139-149. doi: 10.1016/j.bpj.2018.05.035.
Tracking Brownian particles is often employed to map the energy landscape they explore. Such measurements have been exploited to study many biological processes and interactions in soft materials. Yet video tracking is irremediably contaminated by localization errors originating from two imaging artifacts: the "static" errors come from signal noise, and the "dynamic" errors arise from the motion blur due to finite frame-acquisition time. We show that these errors result in systematic and nontrivial biases in the measured energy landscapes. We derive a relationship between the true and the measured potential that elucidates, among other aberrations, the presence of false double-well minima in the apparent potentials reported in recent studies. We further assess several canonical trapping and pair-interaction potentials by using our analytically derived results and Brownian dynamics simulations. In particular, we show that the apparent spring stiffness of harmonic potentials (such as optical traps) is increased by dynamic errors but decreased by static errors. Our formula allows for the development of efficient corrections schemes, and we also present in this work a provisional method for reconstructing true potentials from the measured ones.
追踪布朗粒子常被用于绘制它们所探索的能量景观。此类测量方法已被用于研究软物质中的许多生物过程和相互作用。然而,视频追踪不可避免地受到两种成像伪影引起的定位误差的污染:“静态”误差源于信号噪声,而“动态”误差则源于由于有限的帧采集时间导致的运动模糊。我们表明,这些误差会导致测量的能量景观中存在系统且非平凡的偏差。我们推导出了真实势能和测量势能之间的关系,该关系除了其他偏差之外,还阐明了在最近的研究中报告的明显势能中存在虚假的双峰最小值。我们进一步使用我们的分析推导结果和布朗动力学模拟来评估几个典型的捕获和对相互作用势能。特别是,我们表明,动态误差会增加谐波势(如光阱)的表观弹簧刚度,但会减小静态误差。我们的公式允许开发有效的校正方案,我们还在本文中提出了一种从测量势能重建真实势能的暂定方法。