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We present silicon nanowire (SiNW) devices, which were fabricated in a top-down process from silicon-on-insulator (SOI) substrates with a high yield. Electronic characterization proved reliable electronic properties. Here we report the versatile use of the devices in different applications like label-free sensing of DNA hybridization, electronic detection of cellular signals, and detection of proteins. Compared to micro-sized devices, the SiNWs show an improved sensitivity. Analyses of the electronic cell-nanowire contact showed that the important seal resistance is largely increased, which is a prerequisite for highly-sensitive detection of cellular activity. In all applications the SiNW devices show a reliable performance and are highly interesting for future biomedical products.

Substrate-bound gradients expressed in numerous spatio-temporal patterns play a crucial role during the development of complex neural circuits. A deeper understanding of the axon guidance mechanism is provided by studying the effect of a defined substrate-bound cue on a confined neural network. In this study, we constructed a discontinuous substrate-bound gradient to control neuronal cell position, the path of neurite growth, and axon directionality. A variety of gradient patterns, with slight changes in slope, width, and length were designed and fabricated by microcontact printing using laminin/poly-l-lysine (PLL) or PLL alone. The gradients were tested for neurite growth and their impact on axon guidance of embryonic rat cortical neurons. The neurite length was determined and the axon was evaluated by Tau-1 immunostaining. We found that the microgradients of laminin/PLL and PLL directed neurons' adhesion, differentially controlled the neurite growth, and guided up to 84% of the axons. The effect of the protein micropattern on axon guidance and neurite growth depended on the protein and geometric parameters used. Our approach proved to be very successful in guiding axons of single multipolar neurons with very high efficiency. It could thereby be useful to engineer defined neural networks for analyzing signal processing of functional circuits, as well as to unravel fundamental questions of the axon guidance mechanism.

A contactless conductivity detector integrated into a poly(dimethylsiloxane) microchip for electrophoresis is presented. It adopted the simplest configuration of electrodes commonly used in this detection mode for capillary electrophoresis microchips. Although the chip is based on a simple and effective design, it is able to obtain low detection levels due to the low noise of the detection circuit. A circuit based on a lock-in amplifier was designed on printed circuit boards to read out the signal. The property of the detection cell was studied by applying excitation signals of different frequencies and different amplitudes. It was found that the best detection limit could be achieved with a frequency of 50 kHz and amplitude of 20 V. The performance of the detector was demonstrated by successfully separating and detecting several inorganic ions and also a mixture of heavy metal ions. An average detection limit of 0.4 μM was obtained for inorganic cations. This value is significantly improved compared to similar microchip-based detectors. The presented detector could be promising for mass production due to its properties, such as simple construction, high degree of integration, high performance and low cost.

Here we describe a new analytical method for the detection of two influenza A viruses by nitrocellulose membrane and magnetic sensors that employ a special frequency mixing technique. The combination of the nitrocellulose membrane and magnetic bead detection permits a rapid assay procedure and excludes two steps (the development of color and the stop reaction) required for usual immunochemical detection methods such as ELISA. Quantitative virus detection was performed using magnetic beads conjugated with secondary antibody. The results were compared with conventional assay methods and with a dot-blot assay with fluorescence compound (FITC). Under optimum conditions, our new assay procedure is capable of detecting picograms of virus per well. This new method combining the nitrocellulose membrane and magnetic bead detection reduces analytical time and allows stable and repeatable analyses of samples in point-of-care applications.

Beilstein Journal of Nanotechnology 2, 104-109 (2011). doi:10.3762/bjnano.2.12, Published by Beilstein-Institut zur Förderung der Chemischen Wissenschaften, Frankfurt, M.

Low-frequency noise in an electrolyte-insulator-semiconductor (EIS) structure functionalized with multilayers of polyamidoamine (PAMAM) dendrimer and single-walled carbon nanotubes (SWNT) is studied. The noise spectral density exhibits dependence with the power factor of and for the bare and functionalized EIS sensor, respectively. The gate-voltage noise spectral density is practically independent of the pH value of the solution and increases with increasing gate voltage or gate-leakage current. It has been revealed that functionalization of an EIS structure with a PAMAM/SWNTs multilayer leads to an essential reduction of the noise. To interpret the noise behavior in bare and functionalized EIS devices, a gate-current noise model for capacitive EIS structures based on an equivalent flatband-voltage fluctuation concept has been developed.

A simple poly(dimethylsiloxane) (PDMS) microchip was fabricated with an integrated contactless conductivity detector. This microchip consists of a PDMS cover sheet with embedded channels and a glass substrate holding the electrodes. The electrodes are insulated from the microchannel by a PDMS membrane. n-Dodecyl β-D-maltoside was utilized to modify the channel surface to improve its hydrophilicity and to reduce the strong absorption of PDMS. A readout system based on a multiplier was set up for detection, and a commonly used rectifier circuit was added to this system for direct comparison. A mixture of inorganic cations (K+, Na+, Li+) was successfully separated in 100 × 30 μm microchannels and detected by applying a sinusoidal wave with a frequency of 100 kHz and 6 Vp-p. This microchip system was characterized in terms of sensitivity, reproducibility and linearity. It is suitable for mass production due to its robustness, high reproducibility, ease of fabrication and low cost. This work provides a valuable contribution to the realization of low-noise and high-sensitivity portable devices applicable to food and environmental detection fields.

There is a continuously growing scientific and technological interest to develop and improve the application of artificial sensors. Biological components which are capable to transduce neutral signals into specific, robust and reproducible indicators frame an attractive alternative to construct biohybrid sensors. Since naturally "occurring" biosensors are only sparsely compatible with artificial devices, genetic engineering of eukaryotic cells provides an attractive approach, where cells can be tailored such to detect target compounds with exquisite specificity and sensitivity. We have developed the prototype for a single-cell-based anion-selective biohybrid sensor. HEK293 cells were stably transfected with a gene encoding glycine receptor alpha(1) subunits. These cells were employed as transducers for glycine-evoked chloride currents in a concentration-dependent way. Cultured on substrate-integrated micro-devices, anionic membrane currents of cells were monitored extracellularly with field-effect transistors (FETs) and gold microelectrode arrays (MEAs). The results supported predictions of state-of-the-art models for cell-sensor coupling mechanisms and confirmed that extracellularly recorded anion currents cause similar signals, regardless whether obtained with field-effect transistors or microelectrodes. The whole-cell sensor successfully tracked glycine concentrations differing by three orders of magnitude. To our knowledge this contribution for the first time marks the functional characterization of an anion-selective biohybrid sensor.

Flexible and transparent electronic devices possess crucial advantages over conventional silicon based systems for bioelectronic applications since they are able to adapt to nonplanar surfaces, cause less chronic immunoreactivity, and facilitate easy optical inspection. Here, organic electrochemical transistors (OECTs) are embedded in a flexible matrix of polyimide to record cardiac action potentials. The wafer-scale fabricated devices exhibit transconductances (12 mS V-1 ) and drain-source on-to-off current ratios (≈105 ) comparable to state of the art nonflexible and superior to other reported flexible OECTs. The transfer characteristics of the devices are preserved even after experiencing extremely high bending strain and harsh crumpling. A sub-micrometer poly(3,4-ethylenedioxythiophene) doped with poly(styrenesulfonate) layer results in a fast transport of ions between the electrolyte and the polymer channel characterized by a cut-off frequency of 1200 Hz. Excellent device performance is proved by mapping the propagation of cardiac action potentials with high signal-to-noise ratio. These results demonstrate that the electrical performance of flexible OECTs can compete with hard-material-based OECTs and thus potentially be used for in vivo applications.

We report the fabrication of a silicon nanowire (SiNW) transistor array realized in a top-down process on 4" silicon-on-insulator (SOI) wafers. This process is a second, optimized version combining thermal nanoimprint lithography and wet chemical etching to define planar nanowires and contact lines out of the top silicon layer in one etching step. We used ion implantation to reduce series resistance of source and drain contact lines. In total 56 individually addressable SiNWs were aligned in two lines. Wires dimensions were down to 60 nm in width and 40 nm in height having lengths up to 40 μm. Due to an improved processing we obtained almost identical wire structures and electronic properties throughout the 4" wafer enabling direct comparison of signals from one chip. The wires were capped by a microfuidic structure, which was used to administer different analyte solutions. First pH characterization measurements and molecular detection experiments using charged polyelectrolytes (PEs) and poly(D)lysine were successful. In future these devices could be used to observe diffusion of analytes in a time-dependent, spatially- and quantitatively-resolved readout.

Mecular bridged nanocontacts allow direct electrical addressing of electroactive molecules, which is of interest for the development of molecular based electronic devices. In the present paper, the electroactive molecule 6-ferrocenyl-1-hexanethiol (Fc-HT) was integrated into metal―bridge―metal (MBM) junctions assembled in a scanning tunneling microscope (STM) setup. A diode-like behavior was observed from the current/bias (I t /V b ) signal through Au (substrate)/Fc-HT/Au (tip) junction, which presented an asymmetric current response due to the resonant tunneling between metal electrode and ferrocenylthiol molecules. With gate electrode modulation, the enhancement of the tunneling current can be controlled, which allows to tune the direction of the current rectification. Our investigations demonstrated that ferrocenylthiol bridged MBM nanostructure has potential applications in the future design of higher-order heterojunctions components in combination with electrochemical logic gates.

We present a novel approach for large-scale silicon nanowire (SiNW) array fabrication for bioelectronic applications. Nanoimprint lithography was combined with standard CMOS processing on 4 in. SOI wafers in order to produce highly integrated arrays of silicon nanowire field-effect transistors (SiNW-FET). With a very smooth surface due to wet anisotropic etching, SiNW-FET arrays show a good electronic performance with a subthreshold slope of about 85 mV/decade. When applying a front-gate control of the wires via an electrochemical reference electrode, reliable electronic performance inside an electrolyte solution can be achieved. Our SiNW-FET sensors exhibit almost no electronic hysteresis on forward and backward bias sweeps. In this article the fabrication process, electronic and electrochemical characterizations and first biomolecular detection experiments are presented. For biodetection experiments we used a differential readout between molecule-free wires and wires carrying covalently attached biomolecules such as short, single-stranded DNA or biotin. With our SiNW-FET arrays a reliable detection of biomolecular layers can be achieved.

In the present paper, we demonstrate the electrochemical rectification of a redox current which is transferred between redox probes (ferricyanide) in solution and a gold electrode functionalized with the biomolecular redox mediator microperoxidase-11 (MP-11). MP-11 is the redox active, heme-containing domain of the biological electron shuttle cytochrome c (cyt c). In our system, a unidirectional current develops due to selective electron transport from the bio-functionalized electrode to ferricyanide such that MP-11 controls the read-out of our coupled redox system. The electrode was functionalized by adding a monolayer of undecanethiol (UDT) to promote the physisorption of MP-11 and inhibit the direct electron transfer between redox probe and electrode. The relative position of redox donator, mediator, and acceptor equilibrium potentials defines the charge transport and a potential-dependent electrochemical current rectification. The results of our investigations demonstrate that functional building blocks of proteins can be reassembled into new conceptual devices with operation modes deviating from their native function, which could prove highly useful in future design of biosensors and bioelectronic systems.

Little is known about the chemical equilibrium formed between the micellar complex of oleylamine and aurous halide during the synthesis of Au nanoparticles employing an Au(I) precursor. In this paper, we determine for the first time the stability constant of the intermediate oleylamine/Au complex during the first stages of the reaction that produces Au nanoparticles using an amine compound as a reducing agent. We found that the intermediate complex has two stability constants that were dependent on the concentration of AuCl. These results were supported by Fourier transform infrared and mass spectroscopy analysis, which indicated that an unstable secondary amine was produced during the oxidation of oleylamine and reduction of Au(I). Our findings demonstrate that the reduction of Au(I) species promotes substeps of structural changes in the oleylamine molecule. We anticipate our assays to be a starting point for a more comprehensive understanding of the synthesis of Au nanoparticles based on a templateless ...

The transduction of electric signals from cells to electronic devices is mandatory for medical applications such as neuroprostheses and fundamental research on communication in neuronal networks. Here, the use of diamond with its advantages for biological applications as a new material for biohybrid devices for the detection of cell signals is investigated. Using the surface conductivity of hydrogen-terminated single-crystalline diamond substrates, arrays of solution-gate field-effect transistors were fabricated. The characterization of the transistors reveals a good stability in electrolyte solutions for at least 7 days. On these devices, cardiomyocyte-like HL-1 cells as well as human embryonic kidney cells (HEK293), which were stably transfected with potassium channels, are cultured. Both types of cells show healthy growth and good adhesion to the substrate. The diamond transistors are used to detect electrical signals from both types of cells by recording the extracellular potential. For the HL-1 cells, the shape of action potentials can be resolved and the propagation of the signal across the cell layer is visible. Potassium currents of HEK293 cells are activated with the patch-clamp technique in voltage-clamp mode and simultaneously measured with the field-effect transistors. The ion sensitivity of the diamond surface enables the detection of released potassium ions accumulated in the cleft between transistor and cell.

High-resolution low-field nuclear magnetic resonance (NMR) signals of selected liquid samples were recorded using a nitrogen-cooled superconducting quantum interference device (SQUID). The NMR measurements were performed at Larmor frequencies (f L) from 2 Hz to 40 kHz. The natural spectral linewidth of tap water could be measured in magnetic fields below 7 microtesla. To demonstrate the measurement sensitivity and resolution, J-coupling spectra of 2,2,2-trifluoroethanol were recorded at different measurement fields, with signals separated by several hundreds of Hertz. An additional nitrogen-cooled tuned LC-circuit and a signal recovery procedure involving a pi/2 AC pulse were applied in the higher f L region ( >10 kHz) to enhance the signal-to-noise ratio up to one order of magnitude.

The influence of low-frequency magnetic field disturbances on SQUID based low field (LF) nuclear magnetic resonance (NMR) measurements is investigated. Two types of sinusoidal fields, a homogenous field and a linear gradient field, were applied as artificial disturbance sources. The influences on the free induction decay (FID) signals as well as on the spectra are discussed. The homogeneous disturbance field caused a frequency modulation of the FID signal. The measured spectra were found to be in good agreement with calculated traces obtained from a solution to the Bloch equation. The gradient disturbance field yielded an amplitude-modulated FID signal. In both cases, frequency mixing lines were observed. It is shown that for disturbances at the power line frequency and harmonics, the influence on the NMR spectra is negligible.

Transparent polymers are considered as alternative low-cost mold materials in UV nanoimprint lithography (UV-NIL). Here, we demonstrate a nanoimprint process with molds made of rigid polymers novel for this application. These polymer molds are found to show high performance in the patterning with UV-NIL. Sub-50nm structures were fabricated with this process.

We describe the fabrication, electrical and electrochemical characterization of silicon nanowire arrays, which were processed in a top-down approach using combined nanoimprint lithography and wet chemical etching. We used the top silicon layer as contact line and observed an influence of implantation and subsequent annealing of these lines to the device performance. In addition we found a subthreshold slope dependence on wire size. When operated in a liquid environment, wires can be utilized as pH sensors. We characterized the pH sensitivity in the linear range and in the subthreshold operation regime. As a first proof-of-principle experiment for the later use of the sensors in bioassays, we monitored the buildup ofpolyelectrolyte multilayers on the wire surface.

In this work cytochrome c (cyt c), a heme-containing redox protein, was used as a model system for studying general aspects of wiring proteins into conceptual metal–biomolecule–metal nanoparticle junctions. Electron transfer kinetics of cyt c immobilized on Au(111) via MUA spacer has been investigated in the absence and presence of surface-bound gold nanoparticles. The combined effect of size and immobilization time of AuNPs as well as their binding site to cyt c on the tunnelling rate of the protein has been explored. It has been observed that electrostatic binding of 5 nm-in-diameter gold nanoparticles obviously accelerates the electron transfer rate of cyt c. Potential applications of gold nanoparticles as electric contacts in nanodevices as well as their beneficial or adverse health effects have been suggested. (© 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

A method for impedimetric detection of biomolecules with field-effect transistor (FET) arrays is presented. For biomolecular detection, the corresponding molecules such as single-stranded DNA or bovine serum albumine (BSA) were covalently immobilized to the gate structure of 16-channel open-gate FET devices. A micro spotter system was used to site-selectively attach biomolecules to selected channels out of the array. The differential transistor transfer function (TTF) of the FETs with attached biomolecules compared to free FETs was recorded. With this impedimetric technique it was possible to reliably detect biomolecules, because the recordings were not disturbed by the typical long-term drift of the sensors like in potentiometric readout mode. For transistor gates, where DNA sequences of different length were attached, small differences in the TTF spectra were detected. When BSA was covalently immobilized to the FETs clear differences in the TTF spectra were detected, which were independent on buffer pH variations around the isoelectric point of the protein. Based on the results presented in this article it can be concluded, that the TTF method detects passive components of the biomolecules like resistance and capacitance rather than surface charge effects.

In reported low-field nuclear magnetic resonance (NMR) measurements using Superconducting Quantum Interference Device (SQUID) detection, the pre-polarizing magnetic field has been usually oriented orthogonal to the measuring field, B(p) perpendicular B(m). Melton et al. systematically analyzed the consequences of B(p) decay in time after turnoff and showed that this decay should be nonadiabatic. We evaluated our measuring procedure in the light of that analysis, and found good quantitative agreement. It was showed that, when the decay time constant is comparable to the precession period of the magnetization of the sample, M, the optimum procedure is to orient B(p) parallel to B(m) and to apply a pi/2 pulse to flip M, similar as in the case of conventional NMR.

The microelectrode array (MEA) was used to evaluate the cardioprotective effects of adenosine triphosphate sensitive potassium (K(ATP)) channel activation using potassium channel openers (KCOs) on HL-1 cardiomyocytes subjected to acute chemically induced metabolic inhibition. Beat frequency and extracellular action potential (exAP) amplitude were measured in the presence of metabolic inhibitors (sodium azide (NaN(3)) or 2-deoxyglucose (2-DG)) or KCOs (pinacidil (PIN, a cyanoguanidine derivative, activates sarcolemmal K(ATP) channels) or SDZ PCO400 (SDZ, a benzopyran derivative, activates mitochondrial K(ATP) channels)). The protective effects of these KCOs on metabolically inhibited HL-1 cells were subsequently investigated. Signal shapes indicated that NaN(3) and 2-DG reduced the rate of the sodium (Na(+)) influx signal as reflected by a reduction in beat frequency. PIN and SDZ appeared to reduce both rate of depolarization and extent of the Na(+) influx signals. Pre-treating cardiomyocytes with PIN (0.1 mM), but not SDZ, prevented the reduction of beat frequency associated with NaN(3)- or 2-DG-induced metabolic inhibition. The exAP amplitude was not affected by either KCO. The cardioprotective effect of PIN relative to SDZ may be due to the opening of different K(ATP) channels. This metabolic inhibition model on the MEA may provide a stable platform for the study of cardiac pathophysiology in the future.

In many different bioelectronic applications silicon field-effect devices such as transistors or nanowires are used. Usually native or thermally grown silicon oxides serve as interfacing layer to the liquid. For an effective voltage to current conversion of the devices, the main demands for interface layers are low leakage current, low defect density, and high input capacitance. In this article we describe the fabrication and characterization of ultra-thin silicon oxide/high-if material stacks for bioelectronics. A combination of ultra-thin silicon oxide and DyScO 3 revealed the best results. This material stack is particularly interesting for future fabrication of field-effect devices for bioelectronic applications.

Three-dimensional surface reconstructions from high resolution image stacks of biological specimens, observed by confocal microscopy, have changed the perspective of morphological understanding. In the field of cell-cell or cell-substrate interfaces, combining these two techniques leads to new insights yet also creates a tremendous amount of data. In this article, we present a technique to reduce large, multidimensional data sets from confocal microscopy into one single curve: a membrane allocation profile. Reconstructed cells are represented in a three-dimensional surface from image sections of individual cells. We virtually cut segments of the reconstructed cell membrane parallel to the substrate and calculate the surface areas of each segment. The obtained membrane allocation profiles lead to morphological insights and yield an in vivo ratio of attached and free membrane areas without cell fixation. As an example, glass substrates were modified with different proteins (fibronectin, laminin, concavalin A, extracellular matrix gel, and both isomers of poly-lysine) and presented to HEK293 cells to examine differences in cell morphology and adhesion. We proved that proteins on a substrate could increase the attached portion of a cell membrane, facing the modified substrate, from an average of 32% (glass) to 45% (poly-lysine) of the total membrane surface area.

Nanostructured surfaces have recently gained in importance for electrochemical applications because of an enhanced surface area compared to planar substrates. Due to this property, structured substrates are well suited for electrochemical (bio-)sensors, capacitive coupling with electrogenic cells, and other bioelectronic applications. However, the relationship between electrolytically exposed and redox-active surface areas of nanostructured electrodes compared to planar electrodes is still under discussion. Here, we performed a series of comparative studies to elucidate processes taking place at the electrochemically active surface of gold nanopillars. The pillars, approximately 200 nm in height and 50 nm in diameter, were fabricated using template-assisted nanostructuring. The surface area increase compared to planar electrodes was determined by scanning electron microscopy (SEM), and the redox-active surface area of the same sample was derived from cyclovoltammetric studies. We found consistency between the SEM results and the electrochemically active surfaces determined by cyclic voltammetry of immobilized ferrocenylhexanethiol, immobilized cytochrome c, and oxidation/reduction of gold for small scan rates. Similar values were derived from the capacitance measured by cyclic voltammetry, whereas impedimetric measurements revealed values twice as high. Commonly used diffusion-controlled systems, such as hexacyanoferrate, showed a smaller increase of the electroactive surface area. Finally, we conclude that the sterically restricted diffusion of redox-active species leads to an inaccurate determination of the electroactive surface area of nanosized electrodes.

Combining a suitable microfabrication technology with surface modification techniques provides adherent binding sites for growing cells in spatially controlled regions with a nonfouling background. [5] Such biofunctional patterns on a protein-repellent background yield geometrically confined cellular networks on artificial surfaces. This in vitro technique provides a means of systematic manipulation of the extracellular environment for investigating cell‐surface and cell‐ cell interactions. Covalent immobilization of extracellular factors also provides pattern stability for long-term cell-based experiments. [6]

We present a complementary metal oxide semiconductor (CMOS) process with floating gate field-effect transistors (FG FETs) as sensor inputs for bioelectronic applications. In order to achieve a stable passivation of the devices against electrolyte solutions, we included additional fabrication steps and materials into our advanced process. Main changes to the standard CMOS process were the application of titanium silicide for all interconnects, a passivation layer formed by LPCVD deposition of silicon nitride and oxide, and an oxidation step for the sensing areas exposed to the electrolyte at the end of the fabrication process. Different arrays of FG FETs were fabricated. We present extracellular recordings from rat cardiac myocytes and from genetically modified human embryonic kidney (HEK) cells. The extracellular recordings showed a good signal-to-noise ratio. Due to the design, the chips were robust and showed an excellent long-term stability. They were able to withstand several cleaning procedures and subsequent cell cultures. The CMOS process allowed the implementation of simple integrated circuits. With two 5-bit decoders it was possible to address an array of 32 × 32 FG FETs. Our study paves the way for future fabrication of higher integrated chips with a stable and robust operation when used in bioelectronic applications.

Die Entwicklung von diagnostischen Methoden auf der Basis von DNA-Mikroarrays, bekannt als DNA- oder Gen-Chips, wird in Zukunft in vielen Bereichen neue Möglichkeiten eröffnen. Zurzeit basieren die meisten DNA-Chips auf Fluoreszenzmarkierung oder radioaktiver Markierung der zu analysierenden DNA in Verbindung mit entsprechenden Nachweismethoden für die jeweiligen Marker. In diesem Artikel stellen wir eine markerfreie und voll-elektronische Messmethode vor, die auf der Detektion der DNA mittels Feldeffekt-Sensoren beruht.

Electrogenic cells are able to generate electrical signals which can be measured by various invasive electrophysiological methods such as patch-clamp or sharp microelectrode recordings. Growing cells on the surfaces of e.g. metal microelectrodes or field-effect transistors allows the recording of an extracellular component of these signals. For an understanding of such extracellular signals it is mandatory to get detailed topographical as well as electrical information about the cell-sensor interface. In a first approximation, this interface can be described by a flat disk between cell membrane and sensor surface. For a correct description of the signals, the electrodiffusion of ions in this interface is modeled by using the stationary Poisson-Nernst-Planck equations. We solve the equations analytically, and derive expressions for the potential, the ionic charge densities, and the seal resistance. The results provide a method for determining the distance h between sensor surface and cell membrane. For human embryonic kidney cells, we receive h approximately 70 nm. Comparison with literature shows good agreement.

For magnetic immunoassay applications, a novel magnetic reader for quantification of magnetic particle concentrations has been developed. Upon magnetic excitation at two distinct frequencies incident on the sample, the response signal generated at a sum frequency is detected. The low-frequency field component periodically drives the magnetic particles into saturation, which is probed by the high-frequency field. The appearance of frequency mixing lines is highly specific to the nonlinearity of the superparamagnetic particles’ magnetization. The optimization of field coils, differential pickup coil, preamplifier, demodulators and filters are discussed. The output signal is linear for four orders of magnitude in iron concentration.

Field-effect-based capacitive electrolyte-insulator-semiconductor (EIS) sensors have been utilised for the deoxyribonucleic acid (DNA) immobilisation and hybridisation detection as well as for monitoring the layer-by-layer adsorption of polyelectrolytes (anionic poly(sodium 4-styrene sulfonate) (PSS) and cationic poly(allylamine hydrochloride) (PAH)). The EIS sensors with charged macromolecules have been systematically characterised by capacitance-voltage, constant-capacitance, impedance spectroscopy and atomic-force microscopy methods. The effect of the number and polarity of the polyelectrolyte layers on the shift of the capacitance-voltage curves has been investigated. Alternating potential shifts of about 30-90 mV have been observed after the adsorption of each polyanion and polycation layer, respectively. The DNA immobilisation and hybridisation signals were 35-55 and 24-33 mV, respectively. The possible mechanisms for the sensor responses are discussed.

Microelectrode arrays (MEAs) with evenly distributed multiple sensor spots have been designed for specific applications. Using the MEAs, we determined the relative profiles of potassium channel openers (KCOs) on cultured embryonic Sprague-Dawley rat cardiac myocytes. KCO, pinacidil (PIN), cromakalim (CROM), SDZ PCO400 (SDZ), or its vehicle, was added to the myocytes cumulatively. The action potential signal shapes in the presence of PIN and SDZ show that the changes in voltage over time and the magnitudes of the associated voltage change were reduced concentration-dependently. CROM affected sodium influx more than PIN and SDZ. The comparisons of changes in the rate of beating and propagation speed in the presence of KCOs were made using their corresponding pD(2) values (the negative log of EC(50)). All KCOs caused concentration-dependent reductions in the rate of beating and propagation speed, with SDZ being the most potent. In addition to the signal shapes, rate of beating, and propagation speed, the origin of excitation and the excitation pattern inside the culture can be also extracted. The results show that the present system can differentiate the effects of different KCOs on myocytes. It might be possible to utilise the MEA as a means to classify drug action based upon a combined interpretation of the three different datasets gained from the extracellular recordings. The combination of these observations might be used as 'drug signatures' when profiling drugs in the future.

Recording of extracellular signals with planar metal microelectrodes (ME) has already been presented more than 30 years ago. To date, microelectrode array (MEA) systems are able to measure extracellular signals at about 64 sites, simultaneously. This enables monitoring of electrical activity of many cells in a large area. The extracellular recording technique has become a widely used method for neurological, toxicological or pharmacological studies. It already proved its potential to supplement the classical methods in electrophysiology. The interpretation of the recorded signal shapes in order to extract electrophysiological meaningful data--however--is still under discussion. In this article, we analyse the preamplifier circuit for extracellular recording of cardiac myocyte signals. We use a circuit model for the cell-electrode contact including the first amplification stage. In test experiments, we observe different signal shapes, when different shunt resistors are introduced at the input of the preamplifier. According to the frequency spectra of the recordings, we evaluate the transfer function between the source signal and the readout signal. As a result of our studies, an optimum readout electronics for originally, preserved extracellular signal shapes is proposed. Our amplifier design will be most valuable, if the use of small microelectrodes with high input impedances for in vitro as well as for in vivo experiments is desired.

Lab on a chip 15(1), 237-243 (2015). doi:10.1039/C4LC00805G, Published by RSC, Cambridge

As a powerful and versatile scientific instrument, magnetic tweezers have been widely used in biophysical research areas, such as mechanical cell properties and single molecule manipulation. If one wants to steer bead position, the nonlinearity of magnetic properties and the strong position dependence of the magnetic field in most magnetic tweezers lead to quite a challenge in their control. In this article, we report multi-pole electromagnetic tweezers with high permeability cores yielding high force output, good maneuverability, and flexible design. For modeling, we adopted a piece-wise linear dependence of magnetization on field to characterize the magnetic beads. We implemented a bi-linear interpolation of magnetic field in the work space, based on a lookup table obtained from finite element simulation. The electronics and software were custom-made to achieve high performance. In addition, the effects of dimension and defect on structure of magnetic tips also were inspected. In a workspace with size of 0.1 × 0.1 mm(2), a force of up to 400 pN can be applied on a 2.8 μm superparamagnetic bead in any direction within the plane. Because the magnetic particle is always pulled towards a tip, the pulling forces from the pole tips have to be well balanced in order to achieve control of the particle's position. Active video tracking based feedback control is implemented, which is able to work at a speed of up to 1 kHz, yielding good maneuverability of the magnetic beads.

A novel effective bioelectrochemical sensor interface for enzyme biosensors is proposed. The method is based on in situ synthesis of gold nanostructures (5–15 nm) on the thin-film electrode surface using the oleylamine (OA) method, which provides a high-density, stable, electrode interface nanoarchitecture. New method to activate the surface of the OA-stabilized nanostructured electrochemical interface for further functionalization with biomolecules (glucose oxidase enzyme) using Meerwein's salt is proposed. Using this approach a new biosensor for glucose determination with improved analytical characteristics: wide working range of 0.06–18.5 mM with a sensitivity of 22.6 ± 0.5 μA mM− 1 cm− 2, limit of detection 0.02 mM, high reproducibility, and long lifetime (60 d, 93%) was developed. The surface morphology of the electrodes was characterized by scanning electron microscopy (SEM). The electrochemical properties of the interface were studied by cyclic voltammetry and electrochemical impedance spectroscopy using a Fe(II/III) redox couple. The studies revealed an increase in the electroactive surface area and a decrease in the charge transfer resistance following surface activation with Meerwein's reagent. A remarkably enhanced stability and reproducibility of the sensor was achieved using in situ synthesis of gold nanostructures on the electrode surface, while surface activation with Meerwein's salt proved indispensable in achieving an efficient bioelectrochemical interface.

Electronic DNA sensors based on field-effect transistor arrays operating in liquid environment, offer an alternative method for the detection of biomolecular binding events, without the requirement to label the target molecules. These semiconductor devices are sensitive to electrical charge variations that occur at the surface/electrolyte interface. Using such devices, the hybridization reaction of oligonucleotides with complementary single-stranded oligonucleotides, which are immobilized on the oxide surface of the transistor gate, can be detected. The detection principle is based on the intrinsic charge of the nucleic acid molecules in liquid environment. In this article we present a summary of the ‘DNA BioFET’ project, which has been conducted in our group in recent years. We compare an electrostatic and a covalent immobilization protocol for the probe DNA. Most important for reliable signal readout is the use of a differential approach, for which a DNA microarray represents an ideal bioassay. With our FET setup we can either use a chip-to-chip reference with two electrically identical chips, which are coated with different probe sequences. A more sophisticated method, which makes use of the DNA microarray technology, was established with an aligned microspotting system enabling the coating of different channels out of the microarray with different probe sequences. So far our current chip design uses 16 channels enabling first proof-of-principle experiments. The detection method itself, however, offers the possibility to be up scaled to many thousands of sensor spots like currently used in fluorescence based DNA microarray chips. (© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Capacitive EIS (electrolyte–insulator–semiconductor) sensors and ISFETs (ion-sensitive field-effect transistor) have been utilised for the detection of the layer-by-layer adsorption of charged macromolecules. The field-effect-based sensors with the adsorbed multilayers have been characterised by capacitance–voltage, constant-capacitance and constant-voltage-mode methods. The effect of the number and polarity of the polyelectrolyte layers on the shift of the capacitance–voltage curves along the capacitance and voltage axis has been investigated. Alternating potential shifts of about 30–50 mV have been observed after the adsorption of each polyanion and polycation layer, respectively. The possible mechanisms for the sensor response are discussed.

A spatially resolved delivery of substances integrated with cell culture substrates shows promise for application in pharmacological assays, bioanalytical studies on cell signaling pathways and cell-based biosensors, where control over the extracellular biochemical environment with a cellular resolution is desirable. In this work, we studied a biohybrid system where rat embryonic cortical neuronal networks are reconstructed on microstructured silicon chips and interfaced to microfluidics. The design of cell–cell and cell–medium interactions in confined geometries is presented. We developed an aligned microcontact printing technique (AμCP) for poly(lysine)-extracellular matrix proteins on microstructured chips, which allows a high degree of geometrical control over the network architecture and alignment of the neuronal network with the microfluidic features of a substrate. Spatially resolved on-chip delivery of compounds with a cellular resolution is demonstrated by chemical stimulation of patterned rat cortical neurons within a network with a number of solutions of excitatory neurotransmitter glutamate delivered via microfluidics. The combination of the system described with a patch-clamp technique allowed both modulation of the biochemical environment on a cellular level and the monitoring of electrophysiological properties in the reconstructed rat embryonic cortical networks changed by this microenvironment.

Silanization protocols for glass slides and silicon oxide substrates usually include acid rinsing steps to activate the surfaces prior to silanization. In our group, field-effect transistor devices and electrolyte–insulator–semiconductor structures are used to electronically record signals from cells or to detect biomolecular interactions at the solid–liquid interface. A miniaturized, high sensitive, field-effect-based semiconductor device should expose at its input stage just a thin oxide (

Electronic DNA sensors based on field-effect transistor (FET) arrays operating in liquid environments offer an alternative method for the detection of biomolecular binding events without the requirement to label the analyte molecules. In order to obtain reproducible signals from such sensors, the attachment of the probe molecules to the gate area of the transistor chips requires a well-controlled supramolecular architecture. The FET chips used in this work are non-metallized, 8-channel transistor devices with micrometer dimensions of the gate structures, which are encapsulated to be dipped into an analyte solution. Two identical chips were functionalized with DNA catcher molecules of different sequences and read out in a differential measurement approach. In this article, we describe the details of the surface modification and covalent attachment of the catcher DNA. The surface modification was characterized by XPS, Fourier transform infrared spectroscopy (FT-IR), fluorescence microscopy and imaging ellipsometry. Electrical FET recordings of the immobilization and hybridization of DNA molecules are presented.

Understanding the interface between cells or tissues and artificial materials is of critical importance for a broad range of areas. For example, in neurotechnology, the interfaces between neurons and external devices create a link between technical and the nervous systems by stimulating or recording from neural tissue. Here, a more effective interface is required to enhance the electrical characteristics of neuronal recordings and stimulations. Up to now, the lack of a systematic characterization of cell-electrode interaction turns out to be the major bottleneck. In this work, we employed a recently developed surface plasmon microscope (SPM) to monitor in real-time the cell-metal interface and to measure in situ the gap distance of the cleft with the spatial resolution reaching to the optical diffraction limit. The SPM allowed determination of the distance of human embryonic kidney 293 cells cultured on gold surfaces coated with various peptides or proteins without any labeling. This method can dramatically simplify the interaction investigation at metal-living cell interface and should be incorporated into systematic characterization methods.

It is very challenging to study the behavior of neuronal cells in a network due to the multiple connections between the cells. Our idea is then to simplify such a network with a configuration where cells can have just a fixed number of connections in order to create a well-defined and ordered network. Here, we report about guiding primary cortical neurons with three-dimensional gold microspines selectively functionalized with an amino-terminated molecule.

Due to a number of advances in molecular biology, cell and tissue culture in combination with more sensitive methods to transduce biological signals, it has become increasingly feasible to detect unknown toxicity or pharmacological effects by using biological systems which are electrically coupled to micro- or nanoelectrodes or field-effect transistors (FETs). The coupling of biomolecules with electronic devices is demonstrated. In order to identify the contributions of the various cell signals we have investigated the coupling of cardiac myocytes with FETs. On the other side such systems can also be used to study the very basics of distributed information processing by interfacing cultured neuronal networks with microelectronic devices.

A critical evaluation of the possibilities and limitations of the label-free detection of deoxyribonucleic acid (DNA) hybridization by means of field-effect-based devices is discussed. A new DNA-detection method is introduced, which utilizes an ion-sensitive field-effect device as transducer. The upon the DNA hybridization induced redistribution of the ion concentration within the intermolecular spaces and/or the alteration of the ion sensitivity of the device is proposed as detection mechanism. The theoretical calculations predict a substantial change in the average ion concentration within the intermolecular spaces induced upon hybridization that are enough to obtain a detectable sensor signal.