Ivan R. Perch-Nielsen

Measurements of scattered light on a microchip flow cytometer with integrated polymer based optical elements.

Flow cytometry is widely used for analyzing microparticles, such as cells and bacteria. In this paper, we report an innovative microsystem, in which several different optical elements (waveguides, lens and fiber-to-waveguide couplers) are integrated with microfluidic channels to form a complete microchip flow cytometer. All the optical elements, the microfluidic system, and the fiber-to-waveguide couplers were defined in one layer of polymer (SU-8, negative photoresist) by standard photolithography. With only a single mask procedure required, all the fabrication and packaging processes can be finished in one day. Polystyrene beads were measured in the microchip flow cytometer, and three signals (forward scattering, large angle scattering and extinction) were measured simultaneously for each bead. To our knowledge this is the first time forward scattered light and incident light extinction were measured in a microsystem using integrated optics. The microsystem can be applied for analyzing different kinds of particles and cells, and can easily be integrated with other microfluidic components.

Actuation of microfabricated tools using multiple GPC-based counterpropagating-beam traps.

We explore the functionalities of a generalized phase contrast (GPC) -based multiple-beam trapping system for the actuation of various microfabricated SiO2 structures in liquid suspension. The arrays of optical traps are formed using two counterpropagating light fields, each of which is spatially reconfigurable in both cross-sectional geometry and intensity distribution, either in a user-interactive manner or under computer supervision. Design of microtools includes multiple appendages with rounded endings by which optical traps hold and three-dimensionally actuate individual tools. Proof-of-principle demonstrations show the collective and user-coordinated utility of multiple beams for driving microstructured objects. The potential to integrate these optically powered microtools may lead to more complex miniaturized machineries - a closely achievable goal with the real-time reconfigurable optical traps employed in this work.

Real-time interactive 3D manipulation of particles viewed in two orthogonal observation planes.

The generalized phase contrast (GPC) method has been applied to transform a single TEM00 beam into a manifold of counterpropagating-beam traps capable of real-time interactive manipulation of multiple microparticles in three dimensions (3D). This paper reports on the use of low numerical aperture (NA), non-immersion, objective lenses in an implementation of the GPC-based 3D trapping system. Contrary to high-NA based optical tweezers, the GPC trapping system demonstrated here operates with long working distance (>10 mm), and offers a wider manipulation region and a larger field of view for imaging through each of the two opposing objective lenses. As a consequence of the large working distance, simultaneous monitoring of the trapped particles in a second orthogonal observation plane is demonstrated.

Numerical simulation of travelling wave induced electrothermal fluid flow

Many microdevices for manipulating particles and cells use electric fields to produce a motive force on the particles. The movement of particles in non-uniform electric fields is called dielectrophoresis, and the usual method of applying this effect is to pass the particle suspension over a microelectrode structure. If the suspension has a noticeable conductivity, one important side effect is that the electric field drives a substantial conduction current through the fluid, causing localized Joule-heating. The resulting thermal gradient produces local conductivity and permittivity changes in the fluid. dielectrophoretic forces acting upon these pockets of fluid will then produce motion of both the fluid and the particles. This paper presents a numerical solution of the electrical force and the resulting electrothermal driven fluid flow on a travelling wave structure. This common electrode geometry consists of interdigitated electrodes laid down in a long array, with the phase of the applied potential shifted by 90° on each subsequent electrode. The resulting travelling electric field was simulated and the thermal field and electrical body force on the fluid calculated, for devices constructed from two typical materials: silicon and glass. The electrothermal fluid flow in the electrolyte over the electrode array was then numerically simulated. The model predicts that the thermal field depends on the conductivity and applied voltage, but more importantly on the geometry of the system and the material used in the construction of the device. The velocity of the fluid flow depends critically on the same parameters, with slight differences in the thermal field for glass and silicon leading to diametrically opposite flow direction with respect to the travelling field for the two materials. In addition, the imposition of slight external temperature gradients is shown to have a large effect on the fluid flow in the device, under certain conditions leading to a reversal of the fluid flow direction.

2D optical manipulation and assembly of shape-complementary planar microstructures

Optical trapping and manipulation offer great flexibility as a non-contact microassembly tool. Its application to the assembly of microscale building blocks may open new doors for micromachine technology. In this work, we demonstrate all-optical assembly of microscopic puzzle pieces in a fluidic environment using programmable arrays of trapping beams. Identical shape-complimentary pieces are optically fabricated with submicron resolution using two-photon polymerization (2PP) technique. These are efficiently assembled into space-filling tessellations by a multiple-beam optical micromanipulation system. The flexibility of the system allows us to demonstrate both user-interactive and computer-automated modes of serial and parallel assembly of microscale objects with high spatial and angular positioning precision.

Three-dimensional imaging in three-dimensional optical multi-beam micromanipulation.

In this work, we present a method providing real-time, low cost, three-dimensional imaging in a three-dimensional optical micromanipulation system. The three-dimensional imaging system is based on a small form factor LED based projector. The projector is used to dynamically shape the rear illumination light in a counter-propagating beam-trapping setup. This allows us to produce stereoscopic images, from which the human brain can construct a three-dimensional image, or alternatively image analysis can be applied by a computer, thereby obtaining true three-dimensional coordinates in real-time for the trapped objects.

Autonomous and 3D real-time multi-beam manipulation in a microfluidic environment

The Generalized Phase Contrast (GPC) method of optical 3D manipulation has previously been used for controlled spatial manipulation of live biological specimen in real-time. These biological experiments were carried out over a time-span of several hours while an operator intermittently optimized the optical system. Here we present GPC-based optical micromanipulation in a microfluidic system where trapping experiments are computer-automated and thereby capable of running with only limited supervision. The system is able to dynamically detect living yeast cells using a computer-interfaced CCD camera, and respond to this by instantly creating traps at positions of the spotted cells streaming at flow velocities that would be difficult for a human operator to handle. With the added ability to control flow rates, experiments were also carried out to confirm the theoretically predicted axially dependent lateral stiffness of GPC-based optical traps.

Three-dimensional forces in GPC-based counterpropagating-beam traps

We theoretically investigate the three-dimensional (3D) trapping force acting on a microsphere held in a pair of counterpropagating beams produced by the generalized phase contrast (GPC) method. In the case of opposing beams of equal power, we identify the range of beam waist separation s that results in a stable 3D optical potential-well by assessing the dependence of the axial and transverse force curves on s. We also examine how the force curves are influenced by other parameters such as size and refractive index of the microsphere. Aside from force curves of beam tandems with equal powers, we also numerically calculate force curves for cases of beam pairs having disparate relative strengths. These calculations enable us to elucidate the large dynamic range for axial position control of microparticles in GPC-based counterpropagting-beam traps.

GPC-based optical micromanipulation in 3D real-time using a single spatial light modulator

Using a novel dual-beam readout with the generalized phase contrast (GPC) method, a multiple-beam 3D real-time micromanipulation system requiring only one spatial light modulator (SLM) has been realized. A theoretical framework for the new GPC scheme with two parallel illumination beams is presented and corroborated with an experimental demonstration. Three-dimensional arrays of polystyrene microbeads were assembled in the newly described system. The use of air immersion objective lenses with GPC-based optical trapping allowed the simultaneous viewing of the assemblies in two orthogonal bright-field imaging perspectives.

Direct immobilization of DNA probes on non-modified plastics by UV irradiation and integration in microfluidic devices for rapid bioassay

DNA microarrays have become one of the most powerful tools in the field of genomics and medical diagnosis. Recently, there has been increased interest in combining microfluidics with microarrays since this approach offers advantages in terms of portability, reduced analysis time, low consumption of reagents, and increased system integration. Polymers are widely used for microfluidic systems, but fabrication of microarrays on such materials often requires complicated chemical surface modifications, which hinders the integration of microarrays into microfluidic systems. In this paper, we demonstrate that simple UV irradiation can be used to directly immobilize poly(T)poly(C)-tagged DNA oligonucleotide probes on many different types of plastics without any surface modification. On average, five- and fourfold improvement in immobilization and hybridization efficiency have been achieved compared to surface-modified slides with aminated DNA probes. Moreover, the TC tag only costs 30% of the commonly used amino group modifications. Using this microarray fabrication technique, a portable cyclic olefin copolymer biochip containing eight individually addressable microfluidic channels was developed and used for rapid and parallel identification of Avian Influenza Virus by DNA hybridization. The one-step, cost-effective DNA-linking method on non-modified polymers significantly simplifies microarray fabrication procedures and permits great flexibility to plastic material selection, thus making it convenient to integrate microarrays into plastic microfluidic systems.