Continuous dielectrophoretic size-based particle sorting.

Continuous-flow dielectrophoretic (DEP) particle separation based on size is demonstrated in a microfluidic device. Polystyrene microspheres suspended in a neutrally buoyant aqueous solution are used as model particles to study DEP induced by an array of slanted, planar, interdigitated electrodes inside of a soft-lithography microchannel. The E-field gradients from the slanted electrodes impart a net transverse force component on the particles that causes them to "ratchet" across the channel. Over the length of the device, larger particles are deflected more than smaller particles according to the balance of hydrodynamic drag and DEP forces. Consequently, a flow-focused particle suspension containing different-sized particles is fractionated as the beads flow and separate down the length of the device. The flow behavior of spherical particles is modeled, and the total transverse particle displacement in the microfluidic device predicts fourth-order size and voltage and second-order inverse flow rate dependences. The model is verified experimentally for a range of flow rates, particle sizes, and E-field strengths.

Surfactant-enhanced liquid-liquid extraction in microfluidic channels with inline electric-field enhanced coalescence.

Continuous microfluidic liquid-liquid extraction is realized in a microfluidic device by generating emulsions with large interfacial areas for mass transfer, and subsequently breaking these emulsions using electric fields into easily separated segments of immiscible liquids (plugs). The microfluidic device employs insulated electrodes in a potassium hydroxide-etched channel to create large electric fields (100 kV m(-1)) that drive coalescence of the emulsion phase. The result is a transition from disperse to slug flow that can then readily be separated by gravity. Extractions of phenol and p-nitrophenol from an aqueous to hexane-surfactant solution serve as model systems. In addition to the increased surface area in the emulsion, extraction efficiency is enhanced by reverse micelles resulting from the presence of surfactants. The surfactant concentration is varied approximately 1-10 wt% and a general two-parameter model is developed to quantify the extraction behavior and demonstrate the effectiveness of reverse micelle enhanced extraction.

Micromixing of miscible liquids in segmented gas-liquid flow.

We present an integrated microfluidic system that achieves efficient mixing between two miscible liquid streams by introducing a gas phase, forming a segmented gas-liquid (slug) flow, and completely separating the mixed liquid and gas streams in a planar capillary separator. The recirculation motion associated with segmented flow enhances advection in straight microchannels without requiring additional fabrication steps. Instantaneous velocity fields are quantified by microscopic particle image velocimetry (muPIV). Velocities in the direction normal to the channel amount to approximately 30% of the bulk liquid velocity inside a liquid segment. This value depends only weakly on the length of a liquid segment. Spatial concentration fields and the extent of mixing (EOM) are obtained from pulsed-laser fluorescence microscopy and confocal scanning microscopy measurements. The mixing length is reduced 2-3-fold in comparison with previously reported chaotic micromixers that use three-dimensional microchannel networks or patterned walls. Segmented gas-liquid microflows allow mixing times to be varied over several orders of magnitude between milliseconds and second time scales.

A microfluidic electroporation device for cell lysis.

We demonstrate a micro-electroporation device for cell lysis prior to subcellular analysis. Simple circuit models show that electrical lysis method is advantageous because it is selective towards plasma membrane while leaving organelle membrane undamaged. In addition, miniaturization of this concept leads to negligible heat generation and bubble formation. The designed microdevices were fabricated using a combination of photolithography, metal-film deposition, and electroplating. We demonstrate the electro-lysis of human carcinoma cells in these devices to release the subcellular materials.

Catalyst surface characterization in microfabricated reactors using pulse chemisorption.

The metal dispersion of a Pt-Al2O3 catalyst was measured reproducibly using pulse CO chemisorption with 4 mg of sample in a silicon microfabricated packed-bed reactor, demonstrating the applicability of micoreactors for high-throughput catalyst characterization with quantitative comparison.

Infrared spectroscopy for chemically specific sensing in silicon-based microreactors.

Fourier transform infrared (FT-IR) spectroscopy in a multiple internal reflection (MIR) geometry is integrated with silicon-based microreactors to allow detection of a wide range of chemical species while taking advantage of inexpensive batch fabrication techniques applicable to silicon substrates. The microreactors are fabricated in silicon and glass using standard microfabrication and selective etching techniques. The small ( approximately 1 cm side) reactor size provides access to nearly the full mid-IR frequency region with MIR-FT-IR, allowing us to probe both solution-phase and surface-bound chemical transformations. The wide applicability of this approach is demonstrated with two representative test cases: kinetics of acid-catalyzed ethyl acetate hydrolysis and amidization of surface-tethered amine groups.

A microfabricated device for subcellular organelle sorting.

We report a microfabricated field flow fractionation device for continuous separation of subcellular organelles by isoelectric focusing. The microdevice provides fast separation in very small samples while avoiding large voltages and heating effects typically associated with conventional electrophoresis-based devices. The basis of the separation is the presence of membrane proteins that give rise to the effective isoelectric points of the organelles. Simulations of isoelectric focusing of mitochondria in microchannels are used to assess design parameters, such as dimensions and time scales. In addition, a model of Joule heating effects in the microdevice during operation indicates that there is no significant heating, even without active cooling. The device is fabricated using a combination of photolithography, thin-film metal deposition/patterning, and electroplating techniques. We demonstrate that in the microfluidic devices, mitochondria from cultured cells migrate under the influence of an electric field into a focused band in less than 6 min, consistent with model predictions. We also illustrate separation of mitochondria from whole cells and nuclei as well as the separation of two mitochondrial subpopulations. When automated and operated in parallel, these microdevices should facilitate high-throughput analysis in studies requiring separation of organelles.

Microfluidic synthesis of colloidal silica.

We demonstrate the design, fabrication, and operation of microfluidic chemical reactors for the synthesis of colloidal silica particles. Two reactor configurations are examined: laminar flow reactors and segmented flow reactors. We analyze particle sizes and size distributions and examine their change with varying linear flow velocity and mean residence time. Laminar flow reactors are affected by axial dispersion at high linear velocities, thus leading to wide particle size distributions under these conditions. Gas is used to create a segmented flow, consisting liquid plugs separated by inert gas bubbles. The internal recirculation created in the liquid plugs generates mixing, which eliminates the axial dispersion effects associated with laminar flow reactors and produces a narrow size distribution of silica nanoparticles.

A microfabrication-based dynamic array cytometer.

We have developed a microfabricated device for use in parallel luminescent single-cell assays that can sort populations upon the basis of dynamic functional responses to stimuli. This device is composed of a regular array of noncontact single-cell traps. These traps use dielectrophoresis to stably confine cells and hold them against disrupting fluid flows. Using quantitative modeling, we have designed traps with a novel asymmetric extruded-quadrupole geometry. This new trap can be physically arrayed and electrically addressed, enabling our cytometer. Situating an array of these traps in a microchannel, we have introduced cells into the array and demonstrated observation of fluorescent dynamic responses followed by sorting. Such a device has potential for use in investigating functional processes, as revealed by temporal behavior, in large numbers of single cells.