Highly birefringent vaterite microspheres: production, characterization and applications for optical micromanipulation.

This paper reports on a simple synthesis and characterization of highly birefringent vaterite microspheres, which are composed of 20-30 nm sized nanocrystalls. Scanning electron microscopy shows a quite disordered assembly of nanocrystals within the microspheres. However, using optical tweezers, the effective birefringence of the microspheres was measured to be Deltan = 0.06, which compares to Deltan = 0.1 of vaterite single crystals. This suggests a very high orientation of the nanocrystals within the microspheres. A hyperbolic model of the direction of the optical axis throughout the vaterite spherulite best fits the experimental data. Results from polarized light microscopy further confirm the hyperbolic model.

Synthesis and surface modification of birefringent vaterite microspheres.

This paper reports on the synthesis of birefringent vaterite microspheres with narrow size distribution using a seeded growth method. In a post-treatment the microspheres were stabilized and functionalized through coating with a combination of organosilica and silica. The coating vastly enhanced the stability of the vaterite microspheres in biological buffers and allowed the attachment of biomolecules such as DNA or proteins. As an example, streptavidin was attached to the surface of the functionalized microspheres. These results pave the way for the use of birefringent vaterite particles for the micromanipulation of single biological molecules such as DNA or specific proteins in an optical trap capable of exerting and measuring torques. The stabilized birefringent microspheres may also find use for biosensor and biological screening applications.

Constant power optical tweezers with controllable torque.

We describe a means for controlling the spin angular-momentum flux of a laser beam at constant power, without introducing any elliptical or linear polarization. This allows a controllable torque, acting to spin the particle uniformly, to be exerted on a birefringent particle in optical tweezers. The constant power means that transverse and axial trapping, and heating due to absorption, are unaffected by changing the torque. The torque can be computer controlled and rapidly changed. In addition, the lateral trapping is kept constant. Very low torques can be obtained such that rotational Brownian motion of birefringent particles can be observed. This has the potential to greatly extend the quantitative applications of the rotation of birefringent objects in optical tweezers.

The effect of Mie resonances on trapping in optical tweezers.

We calculate trapping forces, trap stiffness and interference effects for spherical particles in optical tweezers using electromagnetic theory. We show the dependence of these on relative refractive index and particle size. We investigate resonance effects, especially in high refractive index particles where interference effects are expected to be strongest. We also show how these simulations can be used to assist in the optimal design of traps.

Forces in optical tweezers with radially and azimuthally polarized trapping beams.

It has been suggested that radially polarized beams can be used to improve the performance of optical tweezers, with reduced scattering force resulting from both the polarization and the dark center of the beam [Opt. Lett. 32, 1839 (2007)]. We calculate the forces on particles in such traps, using rigorous electromagnetic theory, comparing the results with azimuthally polarized beam, circularly polarized LG 01 beams, and Gaussian beams. Our results agree qualitatively with Opt. Lett. 32, 1839 (2007), but differ quantitatively.

Picoliter viscometry using optically rotated particles.

Important aspects in the field of microrheology are studies of the viscosity of fluids within structures with micrometer dimensions and fluid samples where only microliter volumes are available. We have quantitatively investigated the performance and accuracy of a microviscometer based on rotating optical tweezers, which requires as little as one microliter of sample. We have characterized our microviscometer, including effects due to heating, and demonstrated its ability to perform measurements over a large dynamic range of viscosities (at least two orders of magnitude). We have also inserted a probe particle through the membrane of a cell and measured the viscosity of the intramembranous contents. Viscosity measurements of tears have also been made with our microviscometer, which demonstrate its potential use to study unstimulated eye fluid.

Optical torque on microscopic objects.

We outline in general the role and potential areas of application for the use of optical torque in optical tweezers. Optically induced torque is always a result of transfer of angular momentum from light to a particle with conservation of momentum as an underlying principle. Consequently, rotation can be induced by a beam of light that carries angular momentum (AM) or by a beam that carries no AM but where AM is induced in the beam by the particle. First, we analyze some techniques to exert torque with optical tweezers such as dual beam traps. We also discuss the alignment and rotation which is achieved using laser beams carrying intrinsic AM-either spin or orbital AM, or both. We then discuss the types of particles that can be trapped and rotated in such beams such as absorbing or birefringent particles. We present a systematic study of the alignment of particles with respect to the beam axis and the beam's polarization as a way of inducing optical torque by studying crystals of the protein lysozyme. We present the theory behind quantitative measurements of both spin and orbital momentum transfer. Finally, we discuss the applications of rotation in optically driven micromachines, microrheology, flow field measurements, and microfluidics.

Physics of optical tweezers.

We outline the basic principles of optical tweezers as well as the fundamental theory underlying optical tweezers. The optical forces responsible for trapping result from the transfer of momentum from the trapping beam to the particle and are explained in terms of the momenta of incoming and reflected or refracted rays. We also consider the angular momentum flux of the beam in order to understand and explain optical torques. In order to provide a qualitative picture of the trapping, we treat the particle as a weak positive lens and the forces on the lens are shown. However, this representation does not provide quantitative results for the force. We, therefore, present results of applying exact electromagnetic theory to optical trapping. First, we consider a tightly focused laser beam. We give results for trapping of spherical particles and examine the limits of trappability in terms of type and size of the particles. We also study the effect of a particle on the beam. This exact solution reproduces the same qualitative effect as when treating the particle as a lens where changes in the convergence or divergence and in the direction of the trapping beam result in restoring forces acting on the particle. Finally, we review the fundamental theory of optical tweezers.

Collecting single molecules with conventional optical tweezers.

The size of particles that can be trapped in optical tweezers ranges from tens of nanometers to tens of micrometers. This size regime also includes large single molecules. Here we present experiments demonstrating that optical tweezers can be used to collect polyethylene oxide molecules suspended in water. The molecules that accumulate in the focal volume do not aggregate and therefore represent a region of increased molecule concentration, which can be controlled by the trapping potential. We also present a model that relates the change in concentration to the trapping potential. Since many protein molecules have molecular weights for which this method is applicable the effect may be useful in assisting nucleation of protein crystals.

Integrated optomechanical microelements.

We integrate the optical elements required to generate optical orbital angular momentum into a microdevice. This allows the rotation of either naturally occuring microparticles or specially fabricated optical rotors. We use a two photon photopolymerization process to create microscopic diffractive optical elements, customized to a wavelength of choice, which are integrated with micromachines in microfluidic devices. This enables the application of high optical torques with off-the-shelf optical tweezers systems.

Measurement of the index of refraction of single microparticles.

The refractive index of single microparticles is derived from precise measurement and rigorous modeling of the stiffness of a laser trap. We demonstrate the method for particles of four different materials with diameters from 1.6 to 5.2 microm and achieve an accuracy of better than 1%. The method greatly contributes as a new characterization technique because it works best under conditions (small particle size, polydispersion) where other methods, such as absorption spectroscopy, start to fail. Particles need not be transferred to a particular fluid, which prevents particle degradation or alteration common in index matching techniques. Our results also show that advanced modeling of laser traps accurately reproduces experimental reality.

Mechanics of cellular adhesion to artificial artery templates.

We are using polymer templates to grow artificial artery grafts in vivo for the replacement of diseased blood vessels. We have previously shown that adhesion of macrophages to the template starts the graft formation. We present a study of the mechanics of macrophage adhesion to these templates on a single cell and single bond level with optical tweezers. For whole cells, in vitro cell adhesion densities decreased significantly from polymer templates polyethylene to silicone to Tygon (167, 135, and 65 cells/mm(2)). These cell densities were correlated with the graft formation success rate (50%, 25%, and 0%). Single-bond rupture forces at a loading rate of 450 pN/s were quantified by adhesion of trapped 2-microm spheres to macrophages. Rupture force distributions were dominated by nonspecific adhesion (forces <40 pN). On polystyrene, preadsorption of fibronectin or presence of serum proteins in the cell medium significantly enhanced adhesion strength from a mean rupture force of 20 pN to 28 pN or 33 pN, respectively. The enhancement of adhesion by fibronectin and serum is additive (mean rupture force of 43 pN). The fraction of specific binding forces in the presence of serum was similar for polystyrene and polymethyl-methacrylate, but specific binding forces were not observed for silica. Again, we found correlation to in vivo experiments, where the density of adherent cells is higher on polystyrene than on silica templates, and can be further enhanced by fibronectin adsorption. These findings show that in vitro adhesion testing can be used for template optimization and to substitute for in-vivo experiments.

Orientation of optically trapped nonspherical birefringent particles.

While the alignment and rotation of microparticles in optical traps have received increased attention recently, one of the earliest examples has been almost totally neglected--the alignment of particles relative to the beam axis, as opposed to about the beam axis. However, since the alignment torques determine how particles align in a trap, they are directly relevant to practical applications. Lysozyme crystals are an ideal model system to study factors determining the orientation of nonspherical birefringent particles in a trap. Both their size and their aspect ratio can be controlled by the growth parameters, and their regular shape makes computational modeling feasible. We show that both external (shape) and internal (birefringence) anisotropy contribute to the alignment torque. Three-dimensionally trapped elongated objects either align with their long axis parallel or perpendicular to the beam axis depending on their size. The shape-dependent torque can exceed the torque due to birefringence, and can align negative uniaxial particles with their optic axis parallel to the electric field, allowing an application of optical torque about the beam axis.

Measurement of action spectra of light-activated processes.

We report on a new experimental technique suitable for measurement of light-activated processes, such as fluorophore transport. The usefulness of this technique is derived from its capacity to decouple the imaging and activation processes, allowing fluorescent imaging of fluorophore transport at a convenient activation wavelength. We demonstrate the efficiency of this new technique in determination of the action spectrum of the light mediated transport of rhodamine 123 into the parasitic protozoan Giardia duodenalis.

Optical force field mapping in microdevices.

We present a method for characterizing microscopic optical force fields. Two dimensional vector force maps are generated by measuring the optical force applied to a probe particle for a grid of particle positions. The method is used to map out the force field created by the beam from a lensed fiber inside a liquid filled microdevice. We find transverse gradient forces and axial scattering forces on the order of 2 pN per 10 mW laser power which are constant over a considerable axial range (>35 microm). These findings suggest future useful applications of lensed fibers for particle guiding/sorting. The propulsion of a small particle at a constant velocity of 200 microm s(-1) is shown.

Measurement of the total optical angular momentum transfer in optical tweezers.

We describe a way to determine the total angular momentum, both spin and orbital, transferred to a particle trapped in optical tweezers. As an example an LG(02) mode of a laser beam with varying degrees of circular polarisation is used to trap and rotate an elongated particle with a well defined geometry. The method successfully estimates the total optical torque applied to the particle. For this technique, there is no need to measure the viscous drag on the particle, as it is an optical measurement. Therefore, knowledge of the particle's size and shape, as well as the fluid's viscosity, is not required.

Characterization of optically driven fluid stress fields with optical tweezers.

We present a controlled stress microviscometer with applications to complex fluids. It generates and measures microscopic fluid velocity fields, based on dual beam optical tweezers. This allows an investigation of bulk viscous properties and local inhomogeneities at the probe particle surface. The accuracy of the method is demonstrated in water. In a complex fluid model (hyaluronic acid), we observe a strong deviation of the flow field from classical behavior. Knowledge of the deviation together with an optical torque measurement is used to determine the bulk viscosity. Furthermore, we model the observed deviation and derive microscopic parameters.

Time-resolved photoluminescence spectroscopy of ligand-capped PbS nanocrystals.

PbS nanocrystals are synthesized using colloidal techniques and have their surfaces capped with oleic acid. The absorption band edge of the PbS nanocrystals is tuned between 900 and 580 nm. The PbS nanocrystals exhibit tuneable photoluminescence with large non-resonant Stokes shifts of up to 500 meV. The magnitude of the Stokes shift is found to be dependent upon the size of PbS nanocrystals. Time-resolved photoluminescence spectroscopy of the PbS nanocrystals reveals that the photoluminescence has an extraordinarily long lifetime of 1 µs. This long fluorescence lifetime is attributed to the effect of dielectric screening similar to that observed in other IV-VI semiconductor nanocrystals.

Optical microrheology using rotating laser-trapped particles.

We demonstrate an optical system that can apply and accurately measure the torque exerted by the trapping beam on a rotating birefringent probe particle. This allows the viscosity and surface effects within liquid media to be measured quantitatively on a micron-size scale using a trapped rotating spherical probe particle. We use the system to measure the viscosity inside a prototype cellular structure.