DC corral trapping of single nanoparticles and macromolecules in solution.

Progress in sorting, separating, and characterizing ever smaller amounts of chemical and biological material depends on the availability of methods for the controlled interaction with nanoscale and molecular-size objects. Here, we report on the reversible, tunable trapping of single DNA molecules and other charged micro- and nanoparticles in aqueous solution using a direct-current (DC) corral trap setup. The trap consists of a circular, non-conductive void in a metal-coated surface that, when charged, generates an electrostatic potential well in the proximate solution. Our results demonstrate that stable, nanoscale confinement of charged objects is achievable over extended periods of time, that trap stiffness is controlled by the applied voltage, and that simultaneous trapping of multiple objects is feasible. The approach shows great promise for lab-on-a-chip systems and biomedical applications due to its simplicity, scalability, selectivity, and the capability to manipulate single DNA molecules in standard buffer solutions.

Effect of geometry on dielectrophoretic trap stiffness in microparticle trapping.

Dielectrophoresis, an electrokinetic technique, can be used for contactless manipulation of micro- and nano-size particles suspended in a fluid. We present a 3-D microfluidic DEP device with an orthogonal electrode configuration that uses negative dielectrophoresis to trap spherical polystyrene micro-particles. Traps with three different basic geometric shapes, i.e. triangular, square, and circular, and a fixed trap area of around 900 µm2 were investigated to determine the effect of trap shape on dynamics and strength of particle trapping. Effects of trap geometry were quantitatively investigated by means of trap stiffness, with applied electric potentials from 6 VP-P to 10 VP-P at 1 MHz. Analyzing the trap stiffness with a trapped 4.42 µm spherical particle showed that the triangular trap is the strongest, while the square shape trap is the weakest. The trap stiffness grew more than eight times in triangular traps and six times in both square and circular traps when the potential of the applied electric field was increased from 6 VP-P to 10 VP-P at 1 MHz. With the maximum applied potential, i.e. 10 VP-P at 1 MHz, the stiffness of the triangular trap was 60% and 26% stronger than the square and circular trap, respectively. A finite element model of the microfluidic DEP device was developed to numerically compute the DEP force for these trap shapes. The findings from the numerical computation demonstrate good agreement with the experimental analysis. The analysis of three different trap shapes provides important insights to predict trapping location, strength of the trapping zone, and optimized geometry for high throughput particle trapping.

Quantitative analysis of the three-dimensional trap stiffness of a dielectrophoretic corral trap.

Dielectrophoresis is a robust approach for the manipulation and separation of (bio)particles using microfluidic platforms. We developed a dielectrophoretic corral trap in a microfluidic device that utilizes negative dielectrophoresis to capture single spherical polystyrene particles. Circular-shaped micron-size traps were employed inside the device and the three-dimensional trap stiffness (restoring trapping force from equilibrium trapping location) was analyzed using 4.42 µm particles and 1 MHz of an alternating electric field from 6 VP-P to 10 VP-P . The trap stiffness increased exponentially in the x- and y-direction, and linearly in the z-direction. Image analysis of the trapped particle movements revealed that the trap stiffness is increased 608.4, 539.3, and 79.7% by increasing the voltage from 6 VP-P to 10 VP-P in the x-, y-, and z-direction, respectively. The trap stiffness calculated from a finite element simulation of the device confirmed the experimental results. This analysis provides important insights to predict the trapping location, strength of the trapping, and optimum geometry for single particle trapping and its applications such as single-molecule analysis and drug discovery.

Development of a confocal scanning microscope for fluorescence imaging and spectroscopy at variable temperatures.

We developed and tested a confocal scanning optical microscope that fits into a thermally controlled, commercial research cryostat designed for operation from ambient temperature down to below 4 K. The home-built microscope is a fiber-coupled, self-contained instrument based on readily available mechanical and optical components. Its sample module is sealed in a protective stainless steel tube that minimizes vibrations caused by the flow of cryogenic gas. A high numerical aperture microscope objective specifically designed for cryogenic and high-vacuum applications focuses the excitation light onto the sample, while the core of an optical fiber attached to an avalanche photodiode acts as the confocal detection pinhole. The sample is displaced using a piezotube scanner mounted on top of a three-axis, low-temperature nanopositioner assembly for coarse sample positioning. A broadband polarizing cube beam splitter in the emission path allows for polarization-resolved imaging and spectroscopy. Fluorescence excitation scans are acquired with custom-written software that correlates fluorescence photon counts with the output from a high precision wavelength meter, which is part of a narrow-band, tunable dye laser setup. The imaging and spectral data acquisition capabilities of the microscope were confirmed using a variety of samples and excitation wavelengths at temperatures ranging from 5 K to room temperature.

Realistic modeling of the illumination point spread function in confocal scanning optical microscopy.

In this paper we present a rigorous and general theoretical model for the illumination point spread function of a confocal microscope that correctly reproduces the optical setup. The model uses vectorial theory and assumes that monochromatic light with a Gaussian intensity distribution (such as from a laser or a single-mode fiber) is focused by a microscope objective with high numerical aperture and passes through stratified media on its way to the sample. This covers the important practical case of illumination through up to three layers, which is the situation most commonly encountered in biological microscopy (immersion oil, glass coverslip, aqueous sample medium). It also accounts for objectives that are corrected for a certain coverslip thickness and refractive index but operated under non-design conditions. Furthermore, illumination with linearly, circularly, or elliptically polarized light is covered by introducing a Babinet-Soleil compensator into the beam path. The model leads to a set of analytical equations that are readily evaluated. Two-dimensional intensity distributions for particular cases of interest are presented and discussed.

Fabrication of optical tips from photonic crystal fibers.

We present a procedure for fabricating optical tips from photonic crystal fibers which feature a solid core surrounded by a cladding with a hexagonal, multilayer arrangement of air channels running along the length of the fiber. Such optical tips may have unique advantages for the production of near-field optical aperture probes (i.e., metal-coated optical tips with a subwavelength aperture at the tip apex). With both cladding and core made of pure silica, these fibers are fluorescence-free; they support only a single mode over a broad wavelength range (covering the visible and near-infrared spectrum), which makes them useful for multicolor experiments; and they exhibit zero group velocity dispersion at visible wavelengths, which opens up the possibility of femtosecond applications in the near field. Our tip fabrication procedure leads to a sharp, protruding, central tip formed exclusively from the fiber core amidst a regular arrangement of smaller tips from the inner, microstructured region of the cladding. A mechanism for tip formation is proposed based on optical observations at various stages, which explains the self-centering nature of the process.