Rafael J. Taboryski

Microfluidic dissolved oxygen gradient generator biochip as a useful tool in bacterial biofilm studies

A microfluidic chip for generation of gradients of dissolved oxygen was designed, fabricated and tested. The novel way of active oxygen depletion through a gas permeable membrane was applied. Numerical simulations for generation of O(2) gradients were correlated with measured oxygen concentrations. The developed microsystem was used to study growth patterns of the bacterium Pseudomonas aeruginosa in medium with different oxygen concentrations. The results showed that attachment of Pseudomonas aeruginosa to the substrate changed with oxygen concentration. This demonstrates that the device can be used for studies requiring controlled oxygen levels and for future studies of microaerobic and anaerobic conditions.

Characterization of potassium channel modulators with QPatch automated patch-clamp technology: system characteristics and performance.

Planar silicon chips with 1-2-microm etched holes (average resistance: 2.04 +/- 0.02 MOmega in physiological buffer, n = 274) have been developed for patch-clamp recordings of whole-cell currents from cells in suspension. An automated 16-channel parallel screening system, QPatch 16, has been developed using this technology. A single-channel prototype of the QPatch system was used for validation of the patch-clamp chip technology. We present here data on the quality of patch-clamp recordings and from actual drug screening studies of human potassium channels expressed in cultured cell lines. Using Chinese hamster ovary (CHO) and human embryonic kidney cells (HEK), gigaseals of 4.1 +/- 0.4 GOmega (n = 146) and high-quality whole-cell current recordings were obtained from hERG and KCNQ4 potassium channels. Success rates for gigaseal recordings varied from 40 to 95%, and 67% of the whole-cell configurations lasted for >20 min. Cells were maintained in suspension up to 4 h in a cell storage facility that is integrated in the QPatch 16. No decline in patchability was observed during this time course. A series of screens was conducted with known inhibitors of the hERG and KCNQ4 potassium channels. Dose-response relationship characterizations of verapamil and rBeKm-1 blockage of hERG currents provided IC(50) values similar to values reported in the literature.

Upscaling and automation of electrophysiology: toward high throughput screening in ion channel drug discovery.

Effective screening of large compound libraries in ion channel drug discovery requires the development of new electrophysiological techniques with substantially increased throughputs compared to the conventional patch clamp technique. Sophion Bioscience is aiming to meet this challenge by developing two lines of automated patch clamp products, a traditional pipette-based system called Apatchi-1, and a silicon chip-based system QPatch. The degree of automation spans from semi-automation (Apatchi-1) where a trained technician interacts with the system in a limited way, to a complete automation (QPatch 96) where the system works continuously and unattended until screening of a full compound library is completed. The performance of the systems range from medium to high throughputs.

Injection molded chips with integrated conducting polymer electrodes for electroporation of cells

We present the design-concept for an all polymer injection molded single use microfluidic device. The fabricated devices comprise integrated conducting polymer electrodes and Luer fitting ports to allow for liquid and electrical access. A case study of low voltage electroporation of biological cells in suspension is presented. The working principle of the electroporation device is based on a focusing of the electric field by means of a constriction in the flow channel for the cells. We demonstrate the use of AC voltage for electroporation by applying a 1 kHz, ±50 V square pulse train to the electrodes and show delivery of polynucleotide fluorescent dye in 46% of human acute monocytic leukemia cells passing the constriction.

Fabrication of combined-scale nano- and microfluidic polymer systems using a multilevel dry etching, electroplating and molding process

Microfabricated single-cell capture and DNA stretching devices have been produced by injection molding. The fabrication scheme employed deep reactive ion etching in a silicon substrate, electroplating in nickel and molding in cyclic olefin polymer. This work proposes technical solutions to fabrication challenges associated with chip sealing and demolding of polymer high-volume replication methods. UV-assisted thermal bonding was found to ensure a strong seal of the microstructures in the molded part without altering the geometry of the channels. In the DNA stretching device, a low aspect ratio nanoslit (1/200) connecting two larger micro-channels was used to stretch a 168.5 kbp DNA molecule, while in the other device single-HeLa cells were captured against a micro-aperture connecting two larger microfluidic channels. Different dry etching processes have been investigated for the master origination of the cell-capture device. The combination of a modified Bosch process and an isotropic polysilicon etch was found to ensure the ease of demolding by resulting in slightly positively tapered sidewalls with negligible undercut at the mask interface. (Some figures may appear in colour only in the online journal)

A Cassie-like law using triple phase boundary line fractions for faceted droplets on chemically heterogeneous surfaces.

We present experimental contact angle data for surfaces, which were surface-engineered with a hydrophobic micropattern of hexagonal geometry. The chemically heterogeneous surface of the same hexagonal pattern of defects resulted in faceted droplets of hexagonal shape. When measuring the advancing contact angles with a viewing position aligned parallel to rows of defects, we found that an area averaged Cassie-law failed in describing the data. By replacing the area fractions by line fractions of the triple phase boundary line segments in the Cassie equation, we found excellent agreement with data.

Characterization of poly(3,4-ethylenedioxythiophene):tosylate conductive polymer microelectrodes for transmitter detection

In this paper we investigate the physical and electrochemical properties of micropatterned poly(3,4-ethylenedioxythiophene):tosylate (PEDOT:tosylate) microelectrodes for neurochemical detection. PEDOT:tosylate is a promising conductive polymer electrode material for chip-based bioanalytical applications such as capillary electrophoresis, high-performance liquid chromatography, and constant potential amperometry at living cells. Band electrodes with widths down to 3 μm were fabricated on polymer substrates using UV lithographic methods. The electrodes are electrochemically stable in a range between -200 mV and 700 mV vs. Ag/AgCl and show a relatively low resistance. A wide range of transmitters is shown to oxidize readily on the electrodes. Kinetic rate constants and half wave potentials are reported. The capacitance per area was found to be high (1670 ± 130 μF cm(-2)) compared to other thin film microelectrode materials. Finally, we use constant potential amperometry to measure the release of transmitters from a group of PC 12 cells. The results show how the current response decreases for a series of stimulations with high K(+) buffer.

Observation of Supercurrent Enhancement in SNS Junctions by Nonequilibrium Injection into Supercurrent Carrying Bound Andreev States

We report for the first time enhancement of the supercurrent by means of injection in a mesoscopic three terminal planar SNSNS device made of Al on GaAs. When a current is injected from one of the superconducting Al electrodes at an injection bias

The Influence of Structure Heights and Opening Angles of Micro- and Nanocones on the Macroscopic Surface Wetting Properties

V=\Delta(T)/e

Modular microfluidic system as a model of cystic fibrosis airways

, the DC Josephson current between the other two superconducting electrodes has a maximum, giving evidence for an enhancement due to a non-equilibrium injection into bound Andreev states of the underlying semiconductor. The effect persists to temperatures where the equilibrium supercurrent has vanished.