基于电动现象的细胞光镊阱力测量

提出了一种新的细胞光阱力标定的实验方法，利用细胞电泳原理，设计并研制了一套符合用光镊测力的电动样品池系统，用它替代单光镊操作系统中的普通样品池和高精密压电位移驱动平台，可以用电压调控细胞运动速度，系统相对简单、经济．实验测量结果为：在2～18V／cm电压梯度范围内，酵母细胞的运动速度与电压梯度成正线性关系，即在皮牛量级以下，最大光阱力与光镊光源的功率成线性增加关系．测量结果表明，该方法可实现对细胞光阱力的测量，还可用来测量细胞表面电量．     

Pilus retraction powers bacterial twitching motility

Twitching and social gliding motility allow many gram negative bacteria to crawl along surfaces, and are implicated in a wide range of biological functions. Type IV pili (Tfp) are required for twitching and social gliding, but the mechanism by which these filaments promote motility has remained enigmatic. Here we use laser tweezers to show that Tfp forcefully retract. Neisseria gonorrhoeae cells that produce Tfp actively crawl on a glass surface and form adherent microcolonies. When laser tweezers are used to place and hold cells near a microcolony, retractile forces pull the cells toward the microcolony. In quantitative experiments, the Tfp of immobilized bacteria bind to latex beads and retract, pulling beads from the tweezers at forces that can exceed 80 pN. Episodes of retraction terminate with release or breakage of the Tfp tether. Both motility and retraction mediated by Tfp occur at about 1 microm s(-1) and require protein synthesis and function of the PilT protein. Our experiments establish that Tfp filaments retract, generate substantial force and directly mediate cell movement.     

Simultaneous micromanipulation in multiple planes using a self-reconstructing light beam

Optical tweezers are commonly used for manipulating microscopic particles, with applications in cell manipulation, colloid research, manipulation of micromachines and studies of the properties of light beams. Such tweezers work by the transfer of momentum from a tightly focused laser to the particle, which refracts and scatters the light and distorts the profile of the beam. The forces produced by this process cause the particle to be trapped near the beam focus. Conventional tweezers use gaussian light beams, which cannot trap particles in multiple locations more than a few micrometres apart in the axial direction, because of beam distortion by the particle and subsequent strong divergence from the focal plane. Bessel beams, however, do not diverge and, furthermore, if part of the beam is obstructed or distorted the beam reconstructs itself after a characteristic propagation distance. Here we show how this reconstructive property may be utilized within optical tweezers to trap particles in multiple, spatially separated sample cells with a single beam. Owing to the diffractionless nature of the Bessel beam, secondary trapped particles can reside in a second sample cell far removed ( approximately 3 mm) from the first cell. Such tweezers could be used for the simultaneous study of identically prepared ensembles of colloids and biological matter, and potentially offer enhanced control of 'lab-on-a-chip' and optically driven microstructures.     

The bacteriophage straight phi29 portal motor can package DNA against a large internal force

As part of the viral infection cycle, viruses must package their newly replicated genomes for delivery to other host cells. Bacteriophage straight phi29 packages its 6.6-microm long, double-stranded DNA into a 42 x 54 nm capsid by means of a portal complex that hydrolyses ATP. This process is remarkable because entropic, electrostatic and bending energies of the DNA must be overcome to package the DNA to near-crystalline density. Here we use optical tweezers to pull on single DNA molecules as they are packaged, thus demonstrating that the portal complex is a force-generating motor. This motor can work against loads of up to 57 pN on average, making it one of the strongest molecular motors reported to date. Movements of over 5 microm are observed, indicating high processivity. Pauses and slips also occur, particularly at higher forces. We establish the force-velocity relationship of the motor and find that the rate-limiting step of the motor's cycle is force dependent even at low loads. Notably, the packaging rate decreases as the prohead is filled, indicating that an internal force builds up to approximately 50 pN owing to DNA confinement. Our data suggest that this force may be available for initiating the ejection of the DNA from the capsid during infection.     

磁性纳米颗粒的二维光操纵及团聚效应

我们利用暗场显微镜系统地研究了水剂磁性液体中Fe3O4磁性纳米颗粒在高度聚焦激光作用下的动力学过程.实验研究表明:当激光聚焦在样品池中间时,由于受到沿激光入射方向强散射力和吸收力的作用,磁性纳米颗粒会被排开至光斑的外围;当激光聚焦在样品池上表面时,在光力和样品池上玻片的共同作用下,磁性纳米颗粒会被捕获在样品池的上玻片的下表面并发生团聚效应;此外,我们还研究了不同激光功率作用下形成的磁性纳米团簇对入射白光的透过谱的影响,随着入射激光功率的增加,透过磁性液体的白光的透过率会急剧下降,而且透射谱的中心波长会随着入射激光功率的增加往长波方向移动.     

表面等离激元光学力研究进展

当一束光照射在物质上,光子与物质发生动量交换,部分动量转移到物质,等效于对物质产生作用力,称为光学力.这一作用力非常弱,一般在pN甚至更小的量级,但一定条件下,仍足以捕获和操纵纳米、微米尺度的物体.在金属纳米结构中,由于表面等离激元共振效应,诱导的局域电场可以产生增强的光学力,可以在亚波长尺度实现光操纵,并且由此衍生出一个极具吸引力的研究方向——表面等离激元光学力.本文介绍了利用金属纳米结构进行表面等离激元光学力操纵的最新研究进展.     

Bead movement by single kinesin molecules studied with optical tweezers

Kinesin, a mechanoenzyme that couples ATP hydrolysis to movement along microtubules, is thought to power vesicle transport and other forms of microtubule-based motility. Here, microscopic silica beads were precoated with carrier protein, exposed to low concentrations of kinesin, and individually manipulated with a single-beam gradient-force optical particle trap ('optical tweezers') directly onto microtubules. Optical tweezers greatly improved the efficiency of the bead assay, particularly at the lowest kinesin concentrations (corresponding to approximately 1 molecule per bead). Beads incubated with excess kinesin moved smoothly along a microtubule for many micrometres, but beads carrying from 0.17-3 kinesin molecules per bead, moved, on average, only about 1.4 microns and then spontaneously released from the microtubule. Application of the optical trap directly behind such moving beads often pulled them off the microtubule and back into the centre of the trap. This did not occur when a bead was bound by an AMP.PNP-induced rigor linkage, or when beads were propelled by several kinesin molecules. Our results are consistent with a model in which kinesin detaches briefly from the microtubule during a part of each mechanochemical cycle, rather than a model in which kinesin remains bound at all times.     

单光刀与单光镊激光微束系统

提出了构建一种用于细胞非接触操作的激光微束系统。该系统由 Nd:YAG激光束经声压、热膨胀、汽化等综合效应实现的光刀和 He- Ne激光束经光学动力学效应实现的光镊组成。将两激光束耦合到显微镜中 ,实现了生物细胞的捕获、移动、翻转、打孔等一系列操作。在此基础上 ,分析了形成光镊所必需产生梯度力场的条件和形成光刀对能量的要求 ,进行了系统的总体设计、关键部件设计和选择 ,构建了一套激光微束操作实验系统 ,得到了预期的试验结果。在该系统上成功地实现了非接触细胞操作 ,并对染色体进行了切割。     

Studying the interaction between gyrase and DNA using magnetic tweezers

The DNA gyrase of Escherichia coli plays an essential role in the life of this microorganism.It is unique among all topoisomerases because of its ability to introduce negative supercoils into DNA.This study investigated the single molecular interaction of E.coli gyrase with DNA using magnetic tweezers.The results showed that,in the absence of ATP,gyrase weakly binds the G and T segments.The stretched force of 0.7 pN can gradually destroy the binding,whereas that of 5.9 pN directly destroys it.Addition of high concentrations of norfloxacin enhances gyrase binding to both segments,making them adapt to 5.9 pN.DNA gyrase reduces the plectonemic dimension,which was determined by the bacterial enzyme and not by the pull force.Moreover,it has different affinities for positive supercoils,which it prefers,and negative supercoils.The time distribution of the dissociation of gyrase from DNA has a double-exponential form.We herein propose a model to explain this distribution and compare the results with those of other models.     

纳米ZnO改性PVDF超滤膜的性能

 为改善聚偏氟乙烯(PVDF)超滤膜的亲水性,增强其在水处理中的应用能力,采用相转化法制备了纳米ZnO 改性PVDF超滤膜。分析了纳米ZnO 的添加量对膜结构及性能的影响。通过孔隙率测定、接触角测量、扫描电子显微镜、原子力显微镜、材料试验、超滤实验分别对膜的孔隙率、亲水性、微观结构、机械强度、纯水通量、蛋白截留率及水通量恢复率进行表征。结果表明,纳米ZnO 的质量分数为0.2%时,膜的接触角从改性前的76.3°降至63.4°,亲水性得到明显改善;孔隙率由53.4%升至54.1%;拉伸强度由2.09 MPa 升至2.82 MPa;纯水通量、蛋白截留率及水通量恢复率均有一定程度的提高;膜的断面结构规整,指状孔的尺寸较大,膜表面较光滑。     

水溶液中聚苯乙烯小球的单光束光操纵及有序结构的形成

利用单束强聚焦激光对悬浮于水中的聚苯乙烯小球进行了二维的光操纵,并研究了聚苯乙烯小球在光阱中的动力学过程。实验果表明:在切向光梯度力的作用下,聚苯乙烯小球会在样品盒的衬底上发生自组装行为形成二维有序的六角密排的周期性结构。利用这种单光束光组装的方法,在玻璃基底上制备了聚苯乙烯小球二维有序周期性结构。该研究结果将为利用单光强聚焦激光来制备微纳光子多功能器件提供理论依据及实验参考。     

光阱pN量级阱力的流体力学法测量系统

根据流体力学的分析 ,研究并计算了适合光镊操作生物粒子的无限大平板流场特性和流场分布 ,设计并研制了一套符合光镊pN(皮牛 )量级力测量的液体微循环系统和样品池 ,其液体微流量分流器和缓冲器解决了蠕动泵脉冲缺点 .实际测量和光阱力测试结果显示该系统满足光阱pN量级力的测量要求 .     

不同应力路径下砂岩的宏细观力学响应特性

为了研究应力路径对砂岩的宏观、细观力学响应的影响,基于颗粒离散元方法构建了空心圆柱试样并开展了6组不同应力路径的试验,分别研究了砂岩试样在不同应力路径下的宏观应力-应变响应,以及细观法向与切向接触力的分布规律。研究结果表明：当试样内围压相同时,随着外围压的增加,试样破坏时的体积应变也随之增加,即试样的膨胀现象愈加明显;当试样的外围压相同时,随着试样内围压的增加,试样破坏时的体积应变随之先增长后减小。在中主应力系数不为0时,中主应力系数越大,试样的径向应变与环向应变之和的绝对值越大。在内围压相等的情况下,随着外围压的增加,试样的法向接触力增加。当外围压相等,内围压增大时,试样的法向接触力减小。     

光镊--光的力学效应的研究及应用

激光的发明使得光的力学效应走向了实际应用，光镊是光的力学效应的典型事例。简述了光镊技术的原理和特点——无损伤操控功能和微小力的传感器，介绍具有代表性的研究成果，并展望了光的力学效应研究和应用的发展。     

光阱中多个金纳米棒的双光子荧光效应及热熔合过程

本文利用单束、波长对应金纳米棒长轴表面等离子共振的飞秒脉冲激光对多个长度为40 nm,直径为10 nm的金纳米棒颗粒进行了光捕获,系统研究了金纳米棒颗粒在共振激光作用下的双光子荧光及光致热熔合效应.实验结果表明,在光阱捕获过程中金纳米棒颗粒会激发出明显的双光子荧光.当多个金纳米棒被光力捕获在光斑中心时,金纳米棒发生热熔化并熔合成大尺寸的金纳米团簇.利用这种单光束光镊熔合技术,我们在玻璃衬底上制备了二维有序的金纳米团簇阵列.这一研究对利用金纳米棒颗粒来制备微纳光子结构及多功能光子器件等具有重要的指导意义.     

Massively parallel manipulation of single cells and microparticles using optical images

The ability to manipulate biological cells and micrometre-scale particles plays an important role in many biological and colloidal science applications. However, conventional manipulation techniques--including optical tweezers, electrokinetic forces (electrophoresis, dielectrophoresis, travelling-wave dielectrophoresis), magnetic tweezers, acoustic traps and hydrodynamic flows--cannot achieve high resolution and high throughput at the same time. Optical tweezers offer high resolution for trapping single particles, but have a limited manipulation area owing to tight focusing requirements; on the other hand, electrokinetic forces and other mechanisms provide high throughput, but lack the flexibility or the spatial resolution necessary for controlling individual cells. Here we present an optical image-driven dielectrophoresis technique that permits high-resolution patterning of electric fields on a photoconductive surface for manipulating single particles. It requires 100,000 times less optical intensity than optical tweezers. Using an incoherent light source (a light-emitting diode or a halogen lamp) and a digital micromirror spatial light modulator, we have demonstrated parallel manipulation of 15,000 particle traps on a 1.3 x 1.0 mm2 area. With direct optical imaging control, multiple manipulation functions are combined to achieve complex, multi-step manipulation protocols.     

微粒在飞秒激光光场中的受力特性分析

在分析微粒处于连续高斯光束光场和飞秒脉冲激光光场中轴向光阱力特性的基础上,用数值方法计算了两种情况下微粒所受的轴向力,比较了微粒在两种光场中所受轴向力和热效应的异同,导出了飞秒激光光镊实现对微粒捕获的条件．     

Cytoplasmic dynein functions as a gear in response to load

Cytoskeletal molecular motors belonging to the kinesin and dynein families transport cargos (for example, messenger RNA, endosomes, virus) on polymerized linear structures called microtubules in the cell. These 'nanomachines' use energy obtained from ATP hydrolysis to generate force, and move in a step-like manner on microtubules. Dynein has a complex and fundamentally different structure from other motor families. Thus, understanding dynein's force generation can yield new insight into the architecture and function of nanomachines. Here, we use an optical trap to quantify motion of polystyrene beads driven along microtubules by single cytoplasmic dynein motors. Under no load, dynein moves predominantly with a mixture of 24-nm and 32-nm steps. When moving against load applied by an optical trap, dynein can decrease step size to 8 nm and produce force up to 1.1 pN. This correlation between step size and force production is consistent with a molecular gear mechanism. The ability to take smaller but more powerful strokes under load--that is, to shift gears--depends on the availability of ATP. We propose a model whereby the gear is downshifted through load-induced binding of ATP at secondary sites in the dynein head.     

拉盖尔-高斯光束光镊捕获性质

利用T矩阵法研究光镊中微粒大小与入射光束波长相近时,光镊捕获效率与入射光束的阶数、微粒的折射率和尺寸大小的关系.对拉盖尔-高斯光束光镊和高斯光束光镊的轴向和横向捕获效率进行比较.计算结果表明:不同阶数的拉盖尔-高斯光束对微粒捕获效率的影响不同,阶数不超过4的拉盖尔-高斯光束的捕获效率高;微粒半径增加时,拉盖尔-高斯光束的轴向捕获效率逐渐增大,且捕获域也增加,高斯光束的最大捕获效率基本保持不变但捕获域逐渐增大;微粒折射率增加时,拉盖尔-高斯光束和高斯光束的轴向和横向捕获效率均先增加后递减,捕获效率出现了一个峰值,微粒折射率约在1.39~1.69是稳定捕获的最佳数值.     

Tying a molecular knot with optical tweezers.

Filamentous structures are abundant in cells. Relatively rigid filaments, such as microtubules and actin, serve as intracellular scaffolds that support movement and force, and their mechanical properties are crucial to their function in the cell. Some aspects of the behaviour of DNA, meanwhile, depend critically on its flexibility-for example, DNA-binding proteins can induce sharp bends in the helix. The mechanical characterization of such filaments has generally been conducted without controlling the filament shape, by the observation of thermal motions or of the response to external forces or flows. Controlled buckling of a microtubule has been reported, but the analysis of the buckled shape was complicated. Here we report the continuous control of the radius of curvature of a molecular strand by tying a knot in it, using optical tweezers to manipulate the strand's ends. We find that actin filaments break at the knot when the knot diameter falls below 0.4 microm. The pulling force at breakage is around 1 pN, two orders of magnitude smaller than the tensile stress of a straight filament. The flexural rigidity of the filament remained unchanged down to this diameter. We have also knotted a single DNA molecule, opening up the possibility of studying curvature-dependent interactions with associated proteins. We find that the knotted DNA is stronger than actin.