Micro-fabricated Electrodes Research Articles

Impedance Modelling of FET-Based Biosensors

In designing microfabricated electrodes or field-effect DNA biosensors, the response to the oligonucleotides is often quantified in terms of static changes in the sensed current or potential. More recently, changes to the AC impedance of the device have been used for a variety of reasons. We present here a small-signal, physics-based AC model for biological Field-Effect Transistors (BioFETs), used for detecting DNA molecules. Our model includes the kinetics of the ions in the electrolyte, as well as the electrons and holes in the semiconductor. Using finite-element analysis, we are able to model impedance changes in BioFET structures. Such a model can be used as a quantitative indicator for DNA hybridization experiments.

Nanotextured Electrical Immunoassays for Ultrasensitive Protein Detection

ABSTRACTThis paper describes the development of nanomonitors, which are electrical immunoassays for detection of multiple protein biomarkers. These devices are hybrid sensors with micro-fabricated electrode arrays on a silicon substrate, and integrated nanoporous alumina membranes to provide protein confinement and signal amplification. The disease biomarkers C-reactive protein and Myeloperoxidase have been detected by the nanomonitors in ultra-low concentrations. Proteins were detected in pure samples, human serum, and patient blood samples. The detection accuracy and sensitivity of the nanomonitors in patient samples was comparable to the Enzyme Linked Immunosorbent Assay (ELISA) method of protein detection. Nanomonitors provide the additional benefits of being rapid, label-free, sensitive, and cost effective, providing improvements over traditional protein detection methods, and having potential applications in disease diagnosis.

Mechanical properties of mammalian cells in suspension measured by electro-deformation

We describe a planar, micro-fabricated device for generating fringing non-uniform electric fields. We used it to measure the mechanical properties of individual mammalian cells in suspension by deforming them in time-varying, non-uniform electric fields. Electrical stresses generated by the planar microelectrodes were used to trap and stretch cells, while cell deformation was observed using optical microscopy. Two distinct cell types were compared after fitting strain data with a three-parameter ‘standard linear solid’ model of visco-elasticity, and with a two-parameter power-law method. Chinese hamster ovary (CHO) cells were approximately twice as stiff as U937 human promonocytes, and CHO cells displayed an elastic behaviour with recovery of initial shape, while U937 strain data bore witness to plastic deformation. Our results demonstrate that electrical stresses generated by micro-fabricated electrodes permit mechanical characterization of distinct mammalian cell types.

Amplified Electrokinetic Response by Concentration Polarization near Nanofluidic Channel

Ion concentration polarization is the fundamental transport phenomenon that occurs near ion-selective membranes, but this important membrane phenomenon has been poorly understood due to theoretical and experimental challenges. Here, we report the first direct measurements of detailed flow and electric potential profiles within and near the depletion region. This work is an important step toward a full characterization of this coupled transport problem. Using microfabricated electrodes integrated with the microfluidic device, we measured and confirmed that the electric field inside an ion depletion region is amplified more than 30-fold compared to outside of the depletion zone due to the highly nonuniform ion concentration distribution along the microchannel. As a result, the electrokinetic motion of both fluid (electroosmosis) and particle (electrophoresis) was significantly amplified. The detailed flow profile within the depletion zone was also measured for the first time by optically tracking photobleached neutral dye molecules. We further showed that the amplified electrokinetic flows generated in this device may be used as a field-controlled, microfluidic fluid pump and switch.

Differential pH measurements of metabolic cellular activity in nl culture volumes using microfabricated iridium oxide electrodes

In this paper we describe a new approach to measure pH differences in microfluidic devices and demonstrated acidification rate measurements in on-chip cell culture systems with nl wells. We use two miniaturized identical iridium oxide (IrOx) thin film electrodes (20 micromx400 microm), one as a quasi-reference electrode, the other as a sensing electrode, placed in two confluent compartments on chip. The IrOx electrodes were deposited onto microfabricated platinum (Pt) electrodes simultaneously using electrodeposition. Incorporating the electrodes into a microfluidic device allowed us to expose each electrode to a different solution with a pH difference of one pH unit maintaining a confluent connection between the electrodes. In this configuration, we obtained a reproducible voltage difference between the two IrOx thin film electrodes, which corresponds to the electrode sensitivities of -70 mV/pH at 22 degrees C. In order to measure the acidification rate of cells in nl cell culture volumes we placed one IrOx thin film electrode in the perfusion channel as a quasi-reference electrode and the other in the cell culture volume. We obtained an acidification rate of 0.19+/-0.02 pH/min for fibroblast cells using a stop flow protocol. These results show that we can use two identical miniaturized microfabricated IrOx electrodes to measure pH differences to monitor the metabolic activity of cell cultures on chip. Furthermore, our approach can also be applied in biosensor or bioanalytical applications.

Measuring surface potential components necessary for transmembrane current computation using microfabricated arrays

This study was designed to test the feasibility of using microfabricated electrodes to record surface potentials with sufficiently fine spatial resolution to measure the potential gradients necessary for improved computation of transmembrane current density. To assess that feasibility, we recorded unipolar electrograms from perfused rabbit right ventricular free wall epicardium (n = 6) using electrode arrays that included 25-microm sensors fabricated onto a flexible substrate with 75-microm interelectrode spacing. Electrode spacing was therefore on the size scale of an individual myocyte. Signal conditioning adjacent to the sensors to control lead noise was achieved by routing traces from the electrodes to the back side of the substrate where buffer amplifiers were located. For comparison, recordings were also made using arrays built from chloridized silver wire electrodes of either 50-microm (fine wire) or 250-microm (coarse wire) diameters. Electrode separations were necessarily wider than with microfabricated arrays. Comparable signal-to-noise ratios (SNRs) of 21.2 +/- 2.2, 32.5 +/- 4.1, and 22.9 +/- 0.7 for electrograms recorded using microfabricated sensors (n = 78), fine wires (n = 78), and coarse wires (n = 78), respectively, were found. High SNRs were maintained in bipolar electrograms assembled using spatial combinations of the unipolar electrograms necessary for the potential gradient measurements and in second-difference electrograms assembled using spatial combinations of the bipolar electrograms necessary for surface Laplacian (SL) measurements. Simulations incorporating a bidomain representation of tissue structure and a two-dimensional network of guinea pig myocytes prescribed following the Luo and Rudy dynamic membrane equations were completed using 12.5-microm spatial resolution to assess contributions of electrode spacing to the potential gradient and SL measurements. In those simulations, increases in electrode separation from 12.5 to 75.0, 237.5, and 875.0 microm, which were separations comparable to the finest available with our microfabricated, fine wire, and coarse wire arrays, led to 10%, 42%, and 81% reductions in maximum potential gradients and 33%, 76%, and 96% reductions in peak-to-peak SLs. Maintenance of comparable SNRs for source electrograms was therefore important because microfabrication provides a highly attractive methods to achieve spatial resolutions necessary for improved computation of transmembrane current density.

Trapping single electrons on liquid helium

Surface-state electrons on liquid helium, localised in quantum dots, have been proposed as condensed matter qubits. We now demonstrate experimentally that small numbers of electrons, including a single isolated electron, can be held in a novel electrostatic trap above the surface of superfluid helium. A potential well is created using microfabricated electrodes in a 5 μm diameter pool of helium. Electrons are injected into the trap from an electron reservoir on a helium microchannel. They are individually detected using a superconducting single-electron transistor (SET) as an electrometer. A Coulomb staircase is observed as electrons leave the trap one-by-one until the trap is empty. A design for a prototype quantum information processor using an array of electron traps on liquid helium is presented.

Counting Individual Trapped Electrons on Liquid Helium

We show that small numbers of electrons, including a single isolated electron, can be held in an electrostatic trap above the surface of superfluid helium. A potential well is created using microfabricated electrodes in a 5 μm diameter pool of helium. Electrons are injected into the trap from an electron reservoir on a helium microchannel. They are individually detected using a superconducting single-electron transistor as an electrometer. A Coulomb staircase is observed as electrons leave the trap one–by–one until the trap is empty. A design for a scalable quantum information processor using an array of electron traps is presented.

Analysis of parasites by electrorotation.

The application of the AC electrokinetic technique of electrorotation for studying eukaryotic parasite transmission stages is reviewed. Electrorotation is a noninvasive technique that utilizes electrically energized microelectrode structures within micro-fluidic chambers to probe the physiological structure of micro-organisms. Application of the technique to the transmission life cycle stages of three separate genera of protozoan parasites, Cryptosporidium, Giardia and Cyclospora, and one nematode genus Ascaris, each of significant public health importance, is described. Standard electrorotation apparatus, consisting of micro-fabricated electrodes in a fluidic chip, quadrature sinusoidal signal generator, microscope and image capture system, was used to study each organism. Spectra of cellular rotation rate were recorded as a function of applied electric field frequency and compared with standardized biological tests, where appropriate, to illustrate the effectiveness and versatility of the electrorotation technique. Electrorotational determination of the viability of individual G. intestinalis cysts, Cryptosporidium parvum and Cyclospora cayetanensis oocysts has been achieved. The sporulation state of Cyclospora cayetanensis oocysts was also readily determined, as was the fertilization state of A. suum ova. Electrorotation is a simple, noninvasive and versatile analytical technique suited to a wide range of particle types and capable of incorporation into integrated Lab-on-a-chip devices.

Electric manipulation of bioparticles and macromolecules on microfabricated electrodes.

Bioparticle separation, bioparticle enrichment, and electric field-mediated immune detection were carried out on microfabricated semiconductor chips utilizing ac and dc electric fields. Microscale separation on a chip surface having an active area of approximately 16 mm2 was demonstrated for a mixture of Bacillus globigii spores and Escherichia coli bacteria. Dielectrophoretic enrichment was performed by collecting target bioparticles from a flow stream in flow cells of 47.5 microL, achieving a 20-fold increase in the concentration of E. coli bacteria from a diluted sample, a 28-fold enrichment for peripheral blood mononuclear cells from red blood cells, and a 30-fold increase in white blood cells from diluted whole blood. The ability to manipulate and collect bioparticles and macromolecules at microfabricated electrodes with ac and dc fields was further illustrated in electric field-mediated immunoassays for analyzing the biological identities of E. coli bacteria and B. globigii spores. According to these results, the electric methods for manipulating bioparticles present themselves as viable techniques for novel biomedical applications in sample preparations and biochemical assays on microelectrode arrays.

Cultivation of cells under strong ac-electric field—differentiation between heating and trans-membrane potential effects

Adherently growing mouse fibroblasts were cultivated for days in highly conductive culture media between micro-fabricated electrodes under strong ac-electric fields. Miniature electrodes have improved heat dissipation which allows the use of media with conductivity of more than 1 S/m at field strengths of up to 100 kV/m. Cell division rates, cell motility, cell viability and physiological parameters such as vesiculation were monitored and the actin and β-tubuline structures of cyto-skeleton were imaged by laser scanning fluorescence. The specific effects of polarisation could be differentiated from unspecific effects such as heating. We estimated the real field strength acting on cells. In the kHz-range, field application was clearly limited by membrane dielectric breakdown while temperature increases were less than 3°C. In the MHz-range, much stronger fields could be applied and heating became the limiting factor. Above an induced trans-membrane potential of 130–150 mV cells no longer proliferated under prolonged field application. In the MHz-range (above 5 MHz) cells could be exposed to surprisingly high field strengths (40 kV/m) for days. Therefore, there is a frequency window (up to several 100 MHz) which can be used for cell positioning, manipulation and characterisation techniques without significant loading of cells.

Dielectrophoretic investigations of sub-micrometre latex spheres

A non-uniform AC electric field induces a motion in polarizable particles, called dielectrophoresis. The force responsible for this motion is governed by the dielectric properties both of the suspending medium and of the particles, as well as the geometry of the field. The dielectrophoretic properties of sub-micrometre latex spheres have been studied using micro-fabricated electrode structures. The electric field geometry for electrodes used in the measurements has been solved using numerical analysis. Measurements of the dielectrophoretic properties of the spheres have been made over a range of medium conductivities and applied field frequencies and strengths. Comparisons between the observed behaviour and that expected from theory are presented.

Moleculer Surgery of DNA Using Enzyme-Immobilized Particles

This paper proposes, and experimentally demonstrates, a novel method of 'molecular surgery' of DNA, where a enzyme-labeled particle or probe is pressed against DNA, which is immobilized on the substrate with known conformation and orientation, to make chemical modifications at arbitrary positions of the DNA molecule. In the method presented in this paper, DNA is subjected to high-intensity high-frequency electric field (≥1MV/m, ≈1MHz) created in micro-fabricated electrodes, so that it is 1) stretched to a straight conformation parallel to the field by the electrostatic orientation effect, and 2) positioned onto a substrate by the dielectrophoresis. Once DNA is thus stretch-and-positioned, one has access to arbitrary position of any one of aligned molecules. A latex particle, 2.8μm in diameter, is labeled with DNA-cutting enzyme, DNaseII, and is laser-manipulated and contacted with DNA. It is observed that DNA is instantaneously cut at the contact point. This result is considered to be the first demonstration of space-resolved biochemical modification.