Oscillatory disturbance in force calibration of optical tweezers

In the calibration of the optical trap stiffness, it is found that there appears an attenuating oscillation as an oscillatory disturbance added to the trapped bead movement, when the scanner is driven by a triangular wave input.An equivalent oscillator model is put forward to explain the mechanism of the oscillatory disturbance. Both the measurements and calculations show that the attenuating oscillation comes from the oscillation of the scanner and the triangular wave drive causes this additional oscillation of the scanner. Furthermore, the analysis indicates that the oscillatory disturbance will become stronger, when the stiffness of the trap increases or the natural frequency of the scanner decreases. We adopt another driving way, i.e. a sinusoidal wave input is used instead of the triangular wave input. Our experiment has verified that in this case the oscillatory disturbance is eliminated completely.     

Fast position measurements with scanning line optical tweezers

Scanning line optical tweezers are a powerful tool for the study of colloidal or biomolecular systems in the low-force regime. We present a fast, high-resolution particle position measurement scheme that extends the capabilities of these instruments into the realm of dynamic measurements. The technique is based on synchronous detection of forward-scattered laser light during a line scan. We demonstrate a position resolution of better than 50 nm for bandwidths of as much as 40 kHz for pairs of microspheres trapped in a flat line potential at center-to-center separations of 1.7-6 microm.     

激光功率对光镊光阱刚度标定的影响

光阱刚度是描述光镊对粒子进行操控的重要力学指标,实际使用过程中会受到激光功率的影响。采用均方位移法及玻尔兹曼统计法对搭建的光镊系统进行光阱刚度的标定,利用图像采集方法进行微粒位移的测量,并对两种方法的测量结果进行了比较。为了提高光阱刚度的标定结果的准确性,分析了光路放大倍数、温度变化对最后标定结果的精度影响。结果表明,两种方法进行标定的结果基本相同;光阱刚度在低激光功率（1 mW ~20 mW）范围时随功率近似线性增加,在高功率情况下（25 mW~60 mW）随功率增加不再线性增加,而是趋于一个饱和值。此外,光路放大倍数标定的精确性对标定的精度影响较大,10%的相对误差时,标定结果产生23%的变化,温度对标定的精度影响较小,0.1 ℃的温度变化导致标定结果0.034%的变化。     

New calibration method for position detector for simultaneous measurements of force constants and local viscosity in optical tweezers

We present a new method to calibrate a quadrant photodiode used as position detector to monitor latex beads trapped in optical tweezers. The method combines the dragging Stoke’s force with the thermal noise analysis (power spectral density (PSD)) of the Brownian motion of the trapped bead. Position detector calibrations used in other similar methods normally utilise a bead attached to the coverslip: the voltage-position calibration factor is found by translating the bead across the waist of a laser beam. The so determined calibration factor is then assumed to be the same when beads are investigated in the optical trap. This procedure presents some drawbacks since attached beads can be affected by proximity effects due to the coverslip glass surface which alter the position sensor response itself. On the contrary, our method is able to provide, simultaneously, the calibration factor, the trap stiffness, and the local viscosity of the medium making use of a single trapped bead.     

远场光钳与近场光钳的发展

以经典光学为基础的光钳技术(又称"光镊")在生物、物理和化学等领域得到了广泛的发展和应用,但是该技术受到高倍显微物镜的尺寸和光学衍射极限等多种因素的制约,从而限制了其进一步发展。而远场光纤光钳和近场光钳技术,从不同方面克服了传统光钳的局限。回顾了传统光钳、远场光纤光钳和近场光钳的发展,着重讨论了各种方法的工作原理、实验方法和技术性能,对这几种光钳技术进行了深入地总结和细致地比较。     

Trap position control in the vicinity of reflecting surfaces in optical tweezers

Shift of the trap position from the laser beam waist of optical tweezers is studied experimentally in the presence of a reflecting surface in the vicinity of the focal plane. A standing wave is formed owing to the interference of waves forming the waist and reflected from the surface. The standing wave is shown to affect significantly the resulting trap position. The distance between the surface and the stable optical trap as a function of the trapped particle size is studied numerically. A new method to stabilize the position of the microparticle relative to the surface is proposed. The localization accuracy is determined by the Brownian fluctuations in optical tweezers and is about 10 nm for effective trap stiffness of 4 × 10^?5 N/m.     

Automated optical analysis of young's fringes— optical autocorrelation

Extraction of displacement or velocity data from laser speckle photography or particle image velocimetry transparencies requires measurement of the spacing and orientation of several thousand Young's fringe patterns. In contrast with classical interferometry, fringes formed in this way have poor visibility and it is generally desirable to compute the two-dimensional Fourier transform in order to extract the fringe frequency. The large amount of data produced makes digital acquisition and image processing computationally intensive and time consuming. This paper introduces an automated processor which utilises a photorefractive crystal of bismuth silicon oxide to perform this analysis in a fast and cost-effective manner.     

Substrate-dependent cell elasticity measured by optical tweezers indentation

In the last decade, cell elasticity has been widely investigated as a potential label free indicator for cellular alteration in different diseases, cancer included. Cell elasticity can be locally measured by pulling membrane tethers, stretching or indenting the cell using optical tweezers. In this paper, we propose a simple approach to perform cell indentation at pN forces by axially moving the cell against a trapped microbead. The elastic modulus is calculated using the Hertz-model. Besides the axial component, the setup also allows us to examine the lateral cell–bead interaction. This technique has been applied to measure the local elasticity of HBL-100 cells, an immortalized human cell line, originally derived from the milk of a woman with no evidence of breast cancer lesions. In addition, we have studied the influence of substrate stiffness on cell elasticity by performing experiments on cells cultured on two substrates, bare and collagen-coated, having different stiffness. The mean value of the cell elastic modulus measured during indentation was 26±9 Pa for the bare substrate, while for the collagen-coated substrate it diminished to 19±7 Pa. The same trend was obtained for the elastic modulus measured during the retraction of the cell: 23±10 Pa and 13±7 Pa, respectively. These results show the cells adapt their stiffness to that of the substrate and demonstrate the potential of this setup for low-force probing of modifications to cell mechanics induced by the surrounding environment (e.g. extracellular matrix or other cells).     

Velocimetry microsensors driven by linearly polarized optical tweezers

We report on a new type of single-point velocimetry microsensor that can be positioned in microfluidic devices by use of optical tweezers. The flag-shaped microsensor is readily made by a low-cost two-photon polymerization technique. At rest the linearly polarized optical tweezer traps the microsensor at the focal point, and the flag-plate gets aligned in the polarization direction. Under a fluid flow, the plate rotates to an equilibrium angle that is used to measure the fluid velocity with a micrometer-size spatial resolution. Experimental results are in good agreement with theoretical calculations of optical and hydrodynamic torques on such a flag-shaped microsensor.     

光镊力影响因素的计算分析

运用基于T矩阵算法的开源光镊计算工具包对可能影响光镊力的微粒尺寸、相对折射率以及光束模式进行了研究,计算结果表明,这三方面因素都会对光镊力产生显著影响,微粒直径与波长相等、相对折射率尽可能大时选择恰当的光束模式能够产生最佳的光镊捕获效果.     

Potential mapping of optical tweezers

Optical tweezers are very often used for measurement of piconewton range forces. Depending on the displacement of the trapped bead, the trap may become stiffer which causes considerable underestimation of the measured force. We have shown, both by theory and experiment, that such a stiffening occurs for beads larger than 0.5 μm in radius. For the first time, we have shown that the displacement at which the stiffening starts is size dependent and that the stiffening starts at higher forces for larger beads. We have shown that for the applications, which simultaneous force measurement and position sensing are on demand (such as biopolymer stretching), mid-range sized (～1.5 μm in radius) beads could be the best choice.     

Nanometer gold-silica composite particles manipulated by optical tweezers

We have studied the behavior of nano or micro size composite particles submitted to optical trapping forces and a comparison was made with homogeneous particles of similar dimension. The forces were measured using the power spectrum signal analysis. Most of the results presented were obtained using a lateral effect position sensitive detector (PSD), which allowed the fluctuations of the particle position in the optical trap to be monitored. A 4-quadrant photodiode was also used for the same purpose. We bring clear experimental evidence that the trapping force was increased by a factor of about 2-3 for composite particles made of a colloidal gold core encapsulated in a silica shell, with respect to homogeneous silica or latex beads. These results were discussed in the frame of the various approaches currently used for modeling optical tweezing forces.     

Polarization-induced stiffness asymmetry of optical tweezers

A tightly focused, linearly polarized laser beam, so-called optical tweezers, is proven to be a useful micromanipulation tool. It is known that there is a stiffness asymmetry in the direction perpendicular to the optical axis inherited from the polarization state of the laser. In this Letter, we report our experimental results of stiffness asymmetry for different bead sizes measured at the optimal trapping condition. We also provide the results of our generalized Lorenz-Mie based calculations, which are in good agreement with our experimental results. We also compare our results with previous reports.     

Multiple-beam optical tweezers generated by the generalized phase-contrast method

We demonstrate the application of the generalized phase-contrast (GPC) method to the implementation of a multiple-beam optical tweezer system. Experimental results show the generation of four optical tweezers from a fixed phase mask in conjunction with a GPC system to trap and hold 1-microm-sized polystyrene beads in solution.     

Phase-transition-like properties of double-beam optical tweezers

We report on double-beam optical tweezers that undergo previously unknown phase-transition-like behavior resulting in the formation of more optical traps than the number of beams used to create them. We classify the optical force fields which produce multiple traps for a double-beam system including the critical behavior. This effect is demonstrated experimentally in orthogonally polarized (noninterfering) dual-beam optical tweezers for a silica particle of 2.32 μm diameter. Phase transitions of multiple beam trapping systems have implications for hopping rates between traps and detection of forces between biomolecules using dual-beam optical tweezers. It is an example of a novel dynamic system with multiple states where force fields undergo a series of sign inversions as a function of parameters such as size and beam separation.     

Rotational control within optical tweezers by use of a rotating aperture

We demonstrate a simplified method of rotational control of objects trapped within optical tweezers that does not require high-order modes, interferometric precision, or computer-controlled optical modulators. Inserting a rectangular aperture into the optical beam results in a focused spot that also has rectangular symmetry. We show that an asymmetric object trapped in the beam has its angular orientation fixed such that rotation of the aperture results in a direct rotation of the particle.     

光镊在生命科学中的应用

由于光镊具有无直接接触、无损伤等诸多优点,且光镊产生的力在皮牛顿量级,正好落在生物大分子相互作用力的范围,所以光镊在生物大分子相互作用测量方面取得了越来越广泛的应用.文章详细介绍了光镊的出现、发展过程以及在生命科学中代表性应用结果.这些结果表明,将光镊应用在生命科学领域,可以揭示或证实许多以前不曾深刻了解的生物大分子的活动规律与机制.     

有限远共轭显微镜光镊设计和误差分析

光镊是研究单分子生物物理特性的独特工具, 因而光镊设备的研发是一个极为重要的课题. 本文根据矩阵光学, 对基于有限远共轭显微镜的光镊操控光路进行计算, 得出了阱位径向操控和轴向操控方程, 并分析了光束调控系统、 共焦系统后置透镜和耦合透镜安装位置误差及物镜轴向位置调整对光镊阱位径向及轴向操控精度的影响. 计算结果显示, 当物镜初级像面和耦合透镜像方焦面完全重合, 光束调控系统和耦合透镜的距离误差对阱位径向和轴向操控精度没有影响. 光镊系统元器件定位不准时, 基于无限远共轭显微镜光镊的阱位径向操控误差和轴向操控误差都小于基于有限远共轭显微镜光镊的阱位径向操控误差和轴向操控误差. 当光镊耦合透镜定位误差控制在小于10 mm时, 基于有限远共轭显微镜光镊的径向和轴向操控误差分别小于5.9%和11.4%, 有限远共轭显微镜仍然存在改造为光镊的价值.本文理论为基于有限远共轭显微镜的光镊设计、改造和操控提供理论和实验指导. 关键词： 光镊 光学设计 矩阵 误差     

光镊在生命科学中的应用

由于光镊具有无直接接触、无损伤等诸多优点,且光镊产生的力在皮牛顿量级,正好落在生物大分子相互作用力的范围,所以光镊在生物大分子相互作用测量方面取得了越来越广泛的应用.文章详细介绍了光镊的出现、发展过程以及在生命科学中代表性应用结果.这些结果表明,将光镊应用在生命科学领域,可以揭示或证实许多以前不曾深刻了解的生物大分子的活动规律与机制.