Generally Applicable Holographic Torque Measurement for Optically Trapped Particles.

We present a method to measure the optical torque applied to particles of arbitrary shape such as micrometer-sized micro-organisms or cells held in an optical trap, inferred from the change of angular momentum of light induced by the particle. All torque components can be determined from a single interference pattern recorded by a camera in the back focal plane of a high-NA condenser lens provided that most of the scattered light is collected. We derive explicit expressions mapping the measured complex field in this plane to the torque components. The required phase is retrieved by an iterative algorithm, using the known position of the optical traps as constraints. The torque pertaining to individual particles is accessible, as well as separate spin or orbital parts of the total torque.

Model-based compensation of pixel crosstalk in liquid crystal spatial light modulators.

Spatial light modulators (SLMs) based on liquid crystals are widely used for wavefront shaping. Their large number of pixels allows one to create complex wavefronts. The crosstalk between neighboring pixels, also known as fringing field effect, however, can lead to strong deviations. The realized wavefront may deviate significantly from the prediction based on the idealized assumption that the response across a pixel is uniform and independent of its neighbors. Detailed numerical simulations of the SLM response based on a full 3D physical model accurately match the measured response and properly model the pixel crosstalk. The full model is then used to validate a simplified model that enables much faster crosstalk evaluation and pattern optimization beyond standard performance. General conclusions on how to minimize crosstalk in liquid crystal on silicon (LCoS) SLM systems are derived, as well as a readily accessible estimation of the amount of fringing in a given SLM.

Acoustic force spectroscopy.

Force spectroscopy has become an indispensable tool to unravel the structural and mechanochemical properties of biomolecules. Here we extend the force spectroscopy toolbox with an acoustic manipulation device that can exert forces from subpiconewtons to hundreds of piconewtons on thousands of biomolecules in parallel, with submillisecond response time and inherent stability. This method can be readily integrated in lab-on-a-chip devices, allowing for cost-effective and massively parallel applications.

Direct measurement of axial optical forces.

Direct measurement of optical forces based on recording the change of momentum between the in- and outgoing light does not have specific requirements on particle size or shape, or on beam shape. Thus this approach overcomes many of the limitations of force measurements based on position measurements, which require frequent calibration. In this work we validate the achievable accuracy for direct force measurements in the axial direction for a single beam optical tweezers setup, based on numerical simulations and experimental investigations of situations, where the true force is known. We find that for typical experimental situations a good accuracy with an error of less than 1 % of the maximum force can be achieved, independent of particle size or refractive index, provided that the total amount of light scattered in the backward direction is also taken into account, which is easy to accomplish experimentally. Due to the inherent particle shape independence of the direct force measurement method, these findings support that it provides accurate results for 3D force measurements for particles of arbitrary shape.

How to use a phase-only spatial light modulator as a color display.

We demonstrate that a parallel aligned liquid crystal on silicon (PA-LCOS) spatial light modulator (SLM) without any attached color mask can be used as a full color display with white light illumination. The method is based on the wavelength dependence of the (voltage controlled) birefringence of the liquid crystal pixels. Modern SLMs offer a wide range over which the birefringence can be modulated, leading (in combination with a linear polarizer) to several intensity modulation periods of a reflected light wave as a function of the applied voltage. Because of dispersion, the oscillation period strongly depends on the wavelength. Thus each voltage applied to an SLM pixel corresponds to another reflected color spectrum. For SLMs with a sufficiently broad tuning range, one obtains a color palette (i.e., a "color lookup-table"), which allows one to display color images. An advantage over standard liquid crystal displays (LCDs), which use color masks in front of the individual pixels, is that the light efficiency and the display resolution are increased by a factor of three.

Speeding up liquid crystal SLMs using overdrive with phase change reduction.

Nematic liquid crystal spatial light modulators (SLMs) with fast switching times and high diffraction efficiency are important to various applications ranging from optical beam steering and adaptive optics to optical tweezers. Here we demonstrate the great benefits that can be derived in terms of speed enhancement without loss of diffraction efficiency from two mutually compatible approaches. The first technique involves the idea of overdrive, that is the calculation of intermediate patterns to speed up the transition to the target phase pattern. The second concerns optimization of the target pattern to reduce the required phase change applied to each pixel, which in addition leads to a substantial reduction of variations in the intensity of the diffracted light during the transition. When these methods are applied together, we observe transition times for the diffracted light fields of about 1 ms, which represents up to a tenfold improvement over current approaches. We experimentally demonstrate the improvements of the approach for applications such as holographic image projection, beam steering and switching, and real-time control loops.

Position clamping in a holographic counterpropagating optical trap.

Optical traps consisting of two counterpropagating, divergent beams of light allow relatively high forces to be exerted along the optical axis by turning off one beam, however the axial stiffness of the trap is generally low due to the lower numerical apertures typically used. Using a high speed spatial light modulator and CMOS camera, we demonstrate 3D servocontrol of a trapped particle, increasing the stiffness from 0.004 to 1.5 µN m(-1). This is achieved in the "macro-tweezers" geometry [Thalhammer, J. Opt. 13, 044024 (2011); Pitzek, Opt. Express 17, 19414 (2009)], which has a much larger field of view and working distance than single-beam tweezers due to its lower numerical aperture requirements. Using a 10×, 0.2 NA objective, active feedback produces a trap with similar effective stiffness to a conventional single-beam gradient trap, of order 1 µN m(-1) in 3D. Our control loop has a round-trip latency of 10 ms, leading to a resonance at 20 Hz. This is sufficient bandwidth to reduce the position fluctuations of a 10 µm bead due to Brownian motion by two orders of magnitude. This approach can be trivially extended to multiple particles, and we show three simultaneously position-clamped beads.

Contrast enhancement in widefield CARS microscopy by tailored phase matching using a spatial light modulator.

We introduce a widefield CARS microscope implementation that uses a spatial light modulator to obtain extremely precise control over the pump/probe-beam incidence geometry, which provides the possibility to enhance the image contrast at specific target resonances by fine-tuning the incidence angles. We show how this technique can be used to optimize the image contrast between objects of different size and to practically eliminate the undesired signal from the solvent that embeds small target specimens. Changing the numerical aperture of the illumination from 1.27 to 1.24 improved the ratio of the signals of 500 nm polystyrene beads and the agarose solvent by about 20 dB.

Controlled orientation and sustained rotation of biological samples in a sono-optical microfluidic device.

In cell biology, recently developed technologies for studying suspended cell clusters, such as organoids or cancer spheroids, hold great promise relative to traditional 2D cell cultures. There is, however, growing awareness that sample confinement, such as fixation on a surface or embedding in a gel, has substantial impact on cell clusters. This creates a need for contact-less tools for 3D manipulation and inspection. This work addresses this demand by presenting a reconfigurable, hybrid sono-optical system for contact-free 3D manipulation and imaging, which is suitable for biological samples up to a few hundreds of micrometers in liquid suspension. In our sono-optical device, three independently addressable MHz transducers, an optically transparent top-transducer for levitation and two side-transducers, provide ultrasound excitation from three orthogonal directions. Steerable holographic optical tweezers give us an additional means of manipulation of the acoustically trapped specimen with high spatial resolution. We demonstrate how to control the reorientation or the spinning of complex samples, for instance for 3D visual inspection or for volumetric reconstruction. Whether continuous rotation or transient reorientation takes place depends on the strength of the acoustic radiation torque, arising from pressure gradients, compared to the acoustic viscous torque, arising from the shear forces at the viscous boundary layer around the particle. Based on numerical simulations and experimental insights, we develop a strategy to achieve a desired alignment or continuous rotation around a chosen axis, by tuning the relative strengths of the transducers and thus adjusting the relative contributions of viscous and radiation torques. The approach is widely applicable, as we discuss in several generic examples, with limitations dictated by size and shape asymmetry of the samples.

Optical mirror trap with a large field of view.

Holographic optical tweezers typically require microscope objectives with high numerical aperture and thus usually suffer from the disadvantage of a small field of view and a small working distance. We experimentally investigate an optical mirror trap that is created after reflection of two holographically shaped collinear beams on a mirror. This approach combines a large field of view and a large working distance with the possibility to manipulate particles in a large size range, since it allows to use a microscope objective with a numerical aperture as low as 0.2. In this work we demonstrate robust optical three-dimensional trapping in a range of 1mm x 1mm x 2mm with particle sizes ranging from 1.4 mum up to 45 mum. The use of spatial light modulator based holographic methods to create the trapping beams allows to simultaneously trap many beads in complex, dynamic configurations. We present measurements that characterize the mirror traps in terms of trap stiffness, maximum trapping force and capture range.

Acoustic force mapping in a hybrid acoustic-optical micromanipulation device supporting high resolution optical imaging.

Many applications in the life-sciences demand non-contact manipulation tools for forceful but nevertheless delicate handling of various types of sample. Moreover, the system should support high-resolution optical imaging. Here we present a hybrid acoustic/optical manipulation system which utilizes a transparent transducer, making it compatible with high-NA imaging in a microfluidic environment. The powerful acoustic trapping within a layered resonator, which is suitable for highly parallel particle handling, is complemented by the flexibility and selectivity of holographic optical tweezers, with the specimens being under high quality optical monitoring at all times. The dual acoustic/optical nature of the system lends itself to optically measure the exact acoustic force map, by means of direct force measurements on an optically trapped particle. For applications with (ultra-)high demand on the precision of the force measurements, the position of the objective used for the high-NA imaging may have significant influence on the acoustic force map in the probe chamber. We have characterized this influence experimentally and the findings were confirmed by model simulations. We show that it is possible to design the chamber and to choose the operating point in such a way as to avoid perturbations due to the objective lens. Moreover, we found that measuring the electrical impedance of the transducer provides an easy indicator for the acoustic resonances.