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In spiral phase contrast (SPC) microscopy the edge-enhancement is typically independent of the helicity of the phase vortex filter. Here we show that for layered specimens containing screw-dislocations, as are e.g. present in mica or some crystallized organic substances, the intensity distribution in the filtered image acquires a dependence on the rotational direction of the filter. This allows one to map the distribution of phase singularities in the topography of the sample, by taking the intensity difference between two images recorded with opposite handedness. For the demonstration of this feature in a microscopy set-up, we encode the vortex filter as a binary off-axis hologram displayed on a spatial light modulator (SLM) placed in a Fourier plane. Using a binary grating, the diffraction efficiencies for the plus and minus first diffraction orders are equal, giving rise to two image waves which travel in different directions and are Fourier filtered with opposite helicity. The corresponding two images can be recorded simultaneously in two separate regions of the camera chip. This enables mapping of dislocations in the sample in a single camera exposure, as was demonstrated for various transparent samples.

Microscopic imaging with a setup consisting of a pseudo-random phase mask, and an open CMOS camera, without an imaging objective, is demonstrated. The pseudo random phase mask acts as a diffuser for an incoming laser beam, scattering a speckle pattern to a CMOS chip, which is recorded once as a reference. A sample which is afterwards inserted somewhere in the optical beam path changes the speckle pattern. A single (non-iterative) image processing step, comparing the modified speckle pattern with the previously recorded one, generates a sharp image of the sample. After a first calibration the method works in real-time and allows quantitative imaging of complex (amplitude and phase) samples in an extended three-dimensional volume. Since no lenses are used, the method is free from lens abberations. Compared to standard inline holography the diffuse sample illumination improves the axial sectioning capability by increasing the effective numerical aperture in the illumination path, and it suppresses the undesired so-called twin images. For demonstration, a high resolution spatial light modulator (SLM) is programmed to act as the pseudo-random phase mask. We show experimental results, imaging microscopic biological samples, e.g. insects, within an extended volume at a distance of 15 cm with a transverse and longitudinal resolution of about 60 μm and 400 μm, respectively.

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.

A method to remove undesired diffraction orders of computer-generated binary phase holograms is demonstrated. Normally, the reconstruction of binary Fourier holograms, made from just two phase levels, results in an undesired inverted image from the minus first diffraction order, which is superposed with the desired one. This can be avoided by reconstructing the hologram with a diffuse light field with a pseudorandom, but known, phase distribution, which is taken into account for the hologram computation. As a consequence, only the desired image is reconstructed, whereas all residual light is dispersed, propagating as a diffuse background wave. The method may be advantageous to employ ferroelectric spatial light modulators as holographic display devices, which can display only binary phase holograms, but which have the advantage of fast switching rates.

Recently we demonstrated the applicability of a holographic method for shaping complex wavefronts to spatial light modulator (SLM) systems. Here we examine the potential of this approach for optical micromanipulation. Since the method allows one to shape both amplitude and phase of a trapping light field independently and thus provides full control over scattering and gradient forces, it extends the possibilities of commonly used holographic tweezers systems. We utilize two cascaded phase-diffractive elements which can actually be display side-by-side on a single programmable phase modulator. Theoretically the obtainable light efficiency is close to 100%, in our case the major practical limitation arises from absorption in the SLM. We present data which demonstrate the ability to create user-defined "light pathways" for microparticles driven by transverse radiation pressure.

Abstract: We propose a non-holographic method to create line traps of arbitrary shape in the sample plane. Setting the phase gradient along theses lines gives control over the transverse forces acting on the confined particles. Phase structures, displayed on a spatial light modulator, are optically processed by a spiral phase filter and imaged onto the object plane of a microscope objective. The resulting bright line structures can be used to trap microparticles. Additionally, they exert transverse scattering forces, which can be exploited for inducing orbital motions or for creating “attracting” or “repelling” points, respectively. We give theoretical and experimental evidence that these scattering forces are proportional to the curvature of the line tweezers. [Copyright &y& Elsevier]