Sorting of micron-sized particles using holographic optical Raman tweezers in aqueous medium

We have constructed a holographic optical tweezers system combined with Raman spectroscopy to sort trapped particles. Our software automatically moves the trapped objects to the measurement positions to obtain individual Raman signals from multiple trapped particles. We performed the sorting by comparing their spectra with the previously measured training dataset using the correlation coefficients. We used yeast cells and polystyrene beads as test particles. This study aims to show that biological particles can be separated using single cell analysis with combined holographic optical tweezers and Raman spectroscopy system.     

Axial optical trapping forces on two particles trapped simultaneously by optical tweezers

Optical tweezers, which utilize radiation pressure to control and manipulate microscopic particles, are used for a large number of applications in biology and colloid science. In most applications a single optical tweezers is used to control one single particle. However, two or more particles can be trapped simultaneously. Although this characteristic has been used in applications, no theoretical analysis of the trapping force or the status of the trapped particles is available to our knowledge. We present our calculation, using a ray optics model, of the axial trapping forces on two rigid particles trapped in optical tweezers. The spherical aberration that results from a mismatch of the refractive indices of oil and water is also considered. The results show that the forces exerted by the optical tweezers on the two particles will cause the two particles to touch each other, and the two particles can be stably trapped at a joint equilibrium point. We also discuss the stability of axial trapping. The calculation will be useful in applications of optical tweezers to trap multiple particles.     

Uni- and Bidirectional Rotation and Speed Control in Chiral Photonic Micromotors Powered by Light

Liquid crystalline polymers are attractive materials for untethered miniature soft robots. When they contain azo dyes, they acquire light-responsive actuation properties. However, the manipulation of such photoresponsive polymers at the micrometer scale remains largely unexplored. Here, uni- and bidirectional rotation and speed control of polymerized azo-containing chiral liquid crystalline photonic microparticles powered by light is reported. The rotation of these polymer particles is first studied in an optical trap experimentally and theoretically. The micro-sized polymer particles respond to the handedness of a circularly polarized trapping laser due to their chirality and exhibit uni- and bidirectional rotation depending on their alignment within the optical tweezers. The attained optical torque causes the particles to spin with a rotation rate of several hertz. The angular speed can be controlled by small structural changes, induced by ultraviolet (UV) light absorption. After switching off the UV illumination, the particle recovers its rotation speed. The results provide evidence of uni- and bidirectional motion and speed control in light-responsive polymer particles and offer a new way to devise light-controlled rotary microengines at the micrometer scale.     

Laser Heating Tunability by Off‐Resonant Irradiation of Gold Nanoparticles

Temperature changes in the vicinity of a single absorptive nanostructure caused by local heating have strong implications in technologies such as integrated electronics or biomedicine. Herein, the temperature changes in the vicinity of a single optically trapped spherical Au nanoparticle encapsulated in a thermo‐responsive poly(N‐isopropylacrylamide) shell (Au@pNIPAM) are studied in detail. Individual beads are trapped in a counter‐propagating optical tweezers setup at various laser powers, which allows the overall particle size to be tuned through the phase transition of the thermo‐responsive shell. The experimentally obtained sizes measured at different irradiation powers are compared with average size values obtained by dynamic light scattering (DLS) from an ensemble of beads at different temperatures. The size range and the tendency to shrink upon increasing the laser power in the optical trap or by increasing the temperature for DLS agree with reasonable accuracy for both approaches. Discrepancies are evaluated by means of simple models accounting for variations in the thermal conductivity of the polymer, the viscosity of the aqueous solution and the absorption cross section of the coated Au nanoparticle. These results show that these parameters must be taken into account when considering local laser heating experiments in aqueous solution at the nanoscale. Analysis of the stability of the Au@pNIPAM particles in the trap is also theoretically carried out for different particle sizes.     

Measuring microscopic viscosity with optical tweezers as a confocal probe

We demonstrate, what is to the best our knowledge, a new method for studying the motion of a particle trapped by optical tweezers; in this method the trapping beam itself is used as a confocal probe. By studying the response of the particle to periodic motion of the tweezers, we obtain information about the medium viscosity, particle properties, and trap stiffness. We develop the mathematical model, demonstrate experimentally its validity for our system, and discuss advantages of using this method as a new form of scanning photonic force microscopy for applications in which a high spatial and temporal resolution of the medium viscosity is desired.     

Manipulation of live mouse embryonic stem cells using holographic optical tweezers

We report the ability to move and arrange patterns of live embryonic stem cells using holographic optical tweezers. Single cell suspensions of mouse embryonic stem cells were manipulated with holographic optical tweezers into a variety of patterns including lines, curves and circles. Individual cells were also lifted out of the sample plane highlighting the potential for 3D positional control. Trypan blue dye exclusion and Live/Dead? staining (CMFDA^?1, EthHD^?1) showed that the cells were still viable after manipulation with the optical tweezers. The ability to move individual stem cells into specific, pre-defined patterns provides a method to study how arrangement and associated small-scale interactions occur between neighbouring cells.     

Microparticle trapping in an ultrasonic Bessel beam

This paper describes an acoustic trap consisting of a multi-foci Fresnel lens on 127?μm thick lead zirconate titanate sheet. The multi-foci Fresnel lens was designed to have similar working mechanism to an Axicon lens and generates an acoustic Bessel beam, and has negative axial radiation force capable of trapping one or more microparticle(s). The fabricated acoustic tweezers trapped lipid particles ranging in diameter from 50 to 200?μm and microspheres ranging in diameter from 70 to 90?μm at a distance of 2 to 5?mm from the tweezers without any contact between the transducer and microparticles.     

Optical manipulation in combination with multiphoton microscopy for single-cell studies

We demonstrate how optical tweezers can be incorporated into a multiphoton microscope to achieve three-dimensional imaging of trapped cells. The optical tweezers, formed by a cw 1064 nm Nd:YVO4 laser, were used to trap live yeast cells in suspension while the 4',6-diamidino-2-phenylindole-stained nucleus was imaged in three dimensions by use of a pulsed femtosecond laser. The trapped cell was moved in the axial direction by changing the position of an external lens, which was used to control the divergence of the trapping laser beam. This gives us a simple method to use optical tweezers in the laser scanning of confocal and multiphoton microscopes. It is further shown that the same femtosecond laser as used for the multiphoton imaging could also be used as laser scissors, allowing us to drill holes in the membrane of trapped spermatozoa.     

Mapping the restoring force of an optical trap in three dimensions using a two laser dual-trap approach

A two laser optical tweezers set-up is developed and used to measure deflections of a microsphere trapped in a calibrated spatial light modulator steered probe trap as it is stepped through a three dimensional grid about a fixed test trap. These measurements are used to map the restoring force of the test trap on the microsphere in three dimensions. Results are validated over a common range by comparison to drag force measurements for both silica and polystyrene microspheres.     

Optimization of probe-laser focal offsets for single-particle tracking

In optical tweezers applications, tracking a trapped particle is essential for force measurement. One of the most popular techniques for single-particle tracking is achieved by analyzing the forward and backward light pattern, scattered by the target particle trapped by a trap laser beam, of an additional probe-laser beam with different wavelength whose focus is slightly apart from the trapping center. However, the optimized focal offset has never been discussed. In this paper, we investigate the tracking range and sensitivity as a function of the focal offset between the trapping and the probe-laser beams. As a result, the optimized focal offsets are a 3.3-fold radius ahead and a 2.0-fold radius behind the trapping laser focus in the forward tracking and the backward tracking, respectively. The experimental result agrees well with a theoretical prediction using the Mie scattering theory.     

Single-Microparticle Measurements: Laser Trapping-Absorption Microspectroscopy under Solution-Flow Conditions

A laser trapping-microspectroscopy system combined with a fluid manifold was developed to manipulate and analyze "single" microparticles. A sample solution containing microparticles was introduced to a flow cell set on a microscope stage, and a single particle was trapped by a 1064-nm laser beam. With the particle being trapped, the other particles were pumped out by flowing water to hold the unique microparticle in the flow cell. Under solution-flow conditions, a single microparticle was laser trapped in balance with the gradient (F(g)) and Stokes forces (F(s)) experienced by the particle, and thus, the trapped position was shifted to the downstream side of the 1064-nm laser beam focus. Flow rate and particle size dependencies of this particular positional displacement of the particle were discussed in terms of F(g) and F(s). On the basis of these studies, optical requirements to conduct absorption microspectroscopy of a laser-trapped particle were optimized, and the technique was applied to study a time course of dye adsorption processes in single microparticles. The adsorption rate of Rhodamine B was determined for individual microparticles for the first time.     

Controlled modulation of laser beam and dynamic patterning of colloidal particles using optical tweezers

We present controlled generation of complex-structured beam profiles using diffractive optical element and demonstrate multiple dynamic trapping of colloidal particles. The phase element is programmed to generate various tailored optical fields having structures, similar to that of number three, spiral, and circle but in a tractable manner. Thus, the generated spatially tailored optical fields are confined to focal volume in optical tweezers. This enabled real-time trapping of multiple microscopic objects whereby its transverse organization was controlled in a dynamic manner from one structure to another with the help of spatial light modulator. Such a controlled beam shaping finds potential applications in biophotonics, super resolution imaging, and measurement of biophysical parameters, cell sorting, and micro-manipulation of colloidal particles.     

Three‐Dimensional Exploration and Mechano‐Biophysical Analysis of the Inner Structure of Living Cells

A novel mechanobiological method is presented to explore qualitatively and quantitatively the inside of living biological cells in three dimensions, paving the way to sense intracellular changes during dynamic cellular processes. For this purpose, holographic optical tweezers, which allow the versatile manipulation of nanoscopic and microscopic particles by means of tailored light fields, are combined with self‐interference digital holographic microscopy. This biophotonic holographic workstation enables non‐contact, minimally invasive, flexible, high‐precision optical manipulation and accurate 3D tracking of probe particles that are incorporated by phagocytosis in cells, while simultaneously quantitatively phase imaging the cell morphology. In a first model experiment, internalized polystyrene microspheres with 1 μm diameter are three‐dimensionally moved and tracked in order to quantify distances within the intracellular volume with submicrometer accuracy. Results from investigations on cell swelling provoked by osmotic stimulation demonstrate the homogeneous stretching of the cytoskeleton network, and thus that the proposed method provides a new way for the quantitative 3D analysis of the dynamic intracellular morphology.     

Optical Particle Trapping with Higher-order Doughnut Beams Produced Using High Efficiency Computer Generated Holograms

Abstract

Laser beams containing higher-order phase singularities can be produced with high efficiency computer generated holograms made with very simple equipment. Using such holograms in an optical tweezers experiment we have successfully trapped reflective and absorptive particles in the dark central spot of a focused charge 3 singularity beam. Angular momentum absorbed from the beam can set particles into rotation.     

Using polarization-shaped optical vortex traps for single-cell nanosurgery

Single-cell nanosurgery and the ability to manipulate nanometer-sized subcellular structures with optical tweezers has widespread applications in biology but so far has been limited by difficulties in maintaining the functionality of the transported subcellular organelles. This difficulty arises because of the propensity of optical tweezers to photodamage the trapped object. To address this issue, this paper describes the use of a polarization-shaped optical vortex trap, which exerts less photodamage on the trapped particle than conventional optical tweezers, for carrying out single-cell nanosurgical procedures. This method is also anticipated to find broad use in the trapping of any nanoparticles that are adversely affected by high-intensity laser light.     

Significantly improved trapping lifetime of nanoparticles in an optical trap using feedback control

We demonstrate an increase in trapping lifetime for optically trapped nanoparticles by more than an order of magnitude using feedback control, with no corresponding increase in beam power. Langevin dynamics simulations were used to design the control law, and this technique was then demonstrated experimentally using 100 nm gold particles and 350 nm silica particles. No particle escapes were detected with the controller on, leading to lower limits on the increase in lifetime for 100 nm gold particles of 26 times (at constant average beam power) and 22 times for 350 nm silica particles (with average beam power reduced by one-third). The approach described here can be combined with other techniques, such as counter propagating beams or higher-order optical modes, to trap the smallest nanoparticles and can be used to reduce optical heating of particles that are susceptible to photodamage, such as biological systems.     

Picoliter rheology of gaseous media using a rotating optically trapped birefringent microparticle

An optically trapped birefringent microparticle is rotated by a circularly polarized beam in a confined gaseous medium. By recording the terminal rotation velocity and the change in polarization of the incident trapping beam, we determine the viscosity by probing a picoliter volume of air, carbon dioxide, and argon in the vicinity of the microparticle. We also characterize the optical force acting on a trapped particle in air using the generalized Lorenz-Mie theory taking into account the aberrations present. This opens up a new potential application of optical tweezers for the accurate measurement of gas viscosity in confined geometries.     

基于激光捕捉微粒子实现微细加工的新概念研究

利用高倍显微物镜所聚合的激光在粒子表面所产生的辐射压可捕捉几个微米大小的高强度粒子,这些被捕捉的粒子在激光驱动下,能移动、转动及振动,可成为微加工工具,在被加工表面上形成微米乃至纳米级的表面微细切削。基于几何光学模型对捕捉进行数字仿真,所设计形状粒子的加工力最大可达到mW/N101.210-。     

Design for fully steerable dual-trap optical tweezers

A design for complete beam steering (in three dimensions) of one or two optical tweezers traps is presented. The two most important requirements for efficient and stable movement of an optical trap are identified. A detailed recipe for the construction of a movable optical tweezers trap that fulfills these requirements is given (exemplified with an inverted microscope). The system has been found to allow for precise and free movements of both traps in all three dimensions in a dual-trap optical tweezers configuration and to be robust and reliable, as well as forgiving of small misalignments in the optical system.