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The semiconductor field-effect platform represents a powerful tool for detecting the adsorption and binding of charged macromolecules with direct electrical readout. In this work, a capacitive electrolyte-insulator-semiconductor (EIS) field-effect sensor consisting of an Al-p-Si-SiO2 structure has been applied for real-time in situ electrical monitoring of the layer-by-layer formation of polyelectrolyte (PE) multilayers (PEM). The PEMs were deposited directly onto the SiO2 surface without any precursor layer or drying procedures. Anionic poly(sodium 4-styrene sulfonate) and cationic weak polyelectrolyte poly(allylamine hydrochloride) have been chosen as a model system. The effect of the ionic strength of the solution, polyelectrolyte concentration, number and polarity of the PE layers on the characteristics of the PEM-modified EIS sensors have been studied by means of capacitance-voltage and constant-capacitance methods. In addition, the thickness, surface morphology, roughness and wettabilityof the PE mono- and multilayers have been characterised by ellipsometry, atomic force microscopy and water contact-angle methods, respectively. To explain potential oscillations on the gate surface and signal behaviour of the capacitive field-effect EIS sensor modified with a PEM, a simplified electrostatic model that takes into account the reduced electrostatic screening of PE charges by mobile ions within the PEM has been proposed and discussed.
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We present a label-free method for the detection of DNA hybridization, which is monitored by non-metallized silicon field-effect transistors (FET) in a microarray approach. The described method enables a fast and fully electronic readout of ex situ binding assays. The label-free detection utilizing the field-effect is based on the intrinsic charge of the DNA molecules and/or on changes of the solid-liquid interface impedance, when biomolecules bind to the sensor surface. With our sensor system, usually a time-resolved, dc readout is used. In general, this FET signal suffers from sensor drift, temperature drift, changes in electrolyte composition or pH value, influence of the reference electrode, etc. In this article, we present a differential ac readout concept for FET microarrays, which enables a stable operation of the sensor against many of these side-parameters, reliable readout and a possibility for a quick screening of large sensor arrays. We present the detection of point mutations in short DNA samples with this method in an ex situ binding assay.
Assuntos
Técnicas Biossensoriais/instrumentação , DNA/análise , Polimorfismo de Nucleotídeo Único , Transistores Eletrônicos , Técnicas Biossensoriais/métodos , DNA/genética , Hibridização de Ácido Nucleico , PotenciometriaRESUMO
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.
Assuntos
DNA/química , Capacitância Elétrica , Microscopia de Força Atômica , Hibridização de Ácido Nucleico , Impedância Elétrica , Poliaminas , Polímeros , Ácidos SulfônicosRESUMO
Bonding of polymer-based microfluidics to polymer substrates still poses a challenge for Lab-On-a-Chip applications. Especially, when sensing elements are incorporated, patterned deposition of adhesives with curing at ambient conditions is required. Here, we demonstrate a fabrication method for fully printed microfluidic systems with sensing elements using inkjet and stereolithographic 3D-printing.
Assuntos
Adesivos , Tinta , Dispositivos Lab-On-A-Chip , Técnicas Analíticas Microfluídicas , Impressão , Raios Ultravioleta , Técnicas Analíticas Microfluídicas/instrumentação , Polímeros/químicaRESUMO
Capacitive field-effect sensors modified with a multi-enzyme membrane have been applied for an electronic transduction of biochemical signals processed by enzyme-based AND-Reset and OR-Reset logic gates. The local pH change at the sensor surface induced by the enzymatic reaction was used for the activation of the Reset function for the first time.
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Computadores Moleculares , Enzimas/metabolismo , Lógica , Membranas Artificiais , TransdutoresRESUMO
Insect-based BioFETs (biologically sensitive field-effect transistors) with improved signal characteristics have been developed. These BioFETs require a specifically adapted signal interfacing between a FET as signal transducer and an intact insect antenna as biocomponent. Therefore, different field-effect transistors have been fabricated in order to study the signal transfer at the bioelectronic interface. As relevant features of the BioFET, its current-voltage characteristics, the transconductance and the signal-to-noise ratio have been investigated as affected by the choice of gate insulator materials and gate dimensions (width-to-length ratio, thickness of the dielectric layers). The performance of the improved FET arrangement in the isolated-antenna BioFET was validated by employing dilution series of the plant odour component Z-3-hexen-1-ol.
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Técnicas Biossensoriais , Processamento de Sinais Assistido por Computador , Transistores Eletrônicos , Animais , Calibragem , Besouros , Órgãos dos SentidosRESUMO
A microfluidic chip integrating amperometric enzyme sensors for the detection of glucose, glutamate and glutamine in cell-culture fermentation processes has been developed. The enzymes glucose oxidase, glutamate oxidase and glutaminase were immobilized by means of cross-linking with glutaraldehyde on platinum thin-film electrodes integrated within a microfluidic channel. The biosensor chip was coupled to a flow-injection analysis system for electrochemical characterization of the sensors. The sensors have been characterized in terms of sensitivity, linear working range and detection limit. The sensitivity evaluated from the respective peak areas was 1.47, 3.68 and 0.28 µAs/mM for the glucose, glutamate and glutamine sensor, respectively. The calibration curves were linear up to a concentration of 20 mM glucose and glutamine and up to 10 mM for glutamate. The lower detection limit amounted to be 0.05 mM for the glucose and glutamate sensor, respectively, and 0.1 mM for the glutamine sensor. Experiments in cell-culture medium have demonstrated a good correlation between the glutamate, glutamine and glucose concentrations measured with the chip-based biosensors in a differential-mode and the commercially available instrumentation. The obtained results demonstrate the feasibility of the realized microfluidic biosensor chip for monitoring of bioprocesses.
Assuntos
Técnicas Biossensoriais , Catalase/metabolismo , Análise de Injeção de Fluxo/métodos , Glucose Oxidase/metabolismo , Glutaminase/metabolismo , Técnicas Biossensoriais/economia , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Calibragem , Técnicas Eletroquímicas/economia , Técnicas Eletroquímicas/instrumentação , Eletrodos , Fermentação , Análise de Injeção de Fluxo/economia , Glucose/análise , Ácido Glutâmico/análise , Glutamina/análiseRESUMO
An array of individually addressable nanoplate field-effect capacitive (bio-)chemical sensors based on an SOI (silicon-on-insulator) structure has been developed. The isolation of the individual capacitors was achieved by forming a trench in the top Si layer with a thickness of 350 nm. The realized sensor array allows addressable biasing and electrical readout of multiple nanoplate EISOI (electrolyte-insulator-silicon-on-insulator) capacitive biosensors on the same SOI chip as well as differential-mode measurements. The feasibility of the proposed approach has been demonstrated by realizing sensors for the pH and penicillin concentration detection as well as for the label-free electrical monitoring of polyelectrolyte multilayers formation and DNA (deoxyribonucleic acid)-hybridization event. A potential change of â¼ 120 mV has been registered after the DNA hybridization for the sensor immobilized with perfectly matched single-strand DNA, while practically no signal changes have been observed for a sensor with fully mismatched DNA. The realized examples demonstrate the potential of the nanoplate SOI capacitors as a new basic structural element for the development of different types of field-effect biosensors.
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Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Nanoestruturas/química , Silício , Sequência de Bases , DNA/análise , DNA/genética , Sondas de DNA/genética , Capacitância Elétrica , Técnicas Eletroquímicas , Concentração de Íons de Hidrogênio , Microtecnologia , Nanotecnologia , Hibridização de Ácido Nucleico , Penicilinas/análiseRESUMO
In this article, we report on the electronic monitoring of DNA denaturation by NaOH using electrochemical impedance spectroscopy in combination with fluorescence imaging as a reference technique. The probe DNA consisting of a 36-mer fragment was covalently immobilized on nanocrystalline-diamond electrodes and hybridized with different types of 29-mer target DNA (complementary, single-nucleotide defects at two different positions, and a non-complementary random sequence). The mathematical separation of the impedimetric signals into the time constant for NaOH exposure and the intrinsic denaturation-time constants gives clear evidence that the denaturation times reflect the intrinsic stability of the DNA duplexes. The intrinsic time constants correlate with calculated DNA-melting temperatures. The impedimetric method requires minimal instrumentation, is label-free and fast with a typical time scale of minutes and is highly reproducible. The sensor electrodes can be used repetitively. These elements suggest that the monitoring of chemically induced denaturation at room temperature is an interesting approach to measure DNA duplex stability as an alternative to thermal denaturation at elevated temperatures, used in DNA-melting experiments and single nucleotide polymorphism (SNP) analysis.
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DNA/química , Espectroscopia Dielétrica/métodos , Hibridização de Ácido Nucleico/métodos , DNA/metabolismo , Sondas de DNA/química , Sondas de DNA/metabolismo , Microscopia Confocal , Desnaturação de Ácido Nucleico , Hidróxido de Sódio/química , Temperatura de TransiçãoRESUMO
The integration of living cells together with silicon field-effect devices challenges a new generation of biosensors and bioelectronic devices. Cells are representing highly organised complex systems, optimised by millions of years of evolution and offering a broad spectrum of bioanalytical receptor "tools" such as enzymes, nucleic acids proteins, etc. Their combination with semiconductor-based electronic chips allows the construction of functional hybrid systems with unique functional and electronic properties for both fundamental studies and biosensoric applications. This review article summarises recent advances and trends in research and development of cell/transistor hybrids (cell-based field-effect transistors) as well as light-addressable potentiometric sensors.
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Técnicas Biossensoriais/métodos , Células/metabolismo , Animais , Adesão Celular , Sobrevivência Celular , Humanos , Concentração de Íons de Hidrogênio , Luz , PotenciometriaRESUMO
The feasibility of a capacitive field-effect EDIS (electrolyte-diamond-insulator-semiconductor) platform for multi-parameter sensing is demonstrated by realising EDIS sensors with an O-terminated nanocrystalline-diamond (NCD) film as transducer material for the detection of pH and penicillin concentration as well as for the label-free electrical monitoring of adsorption and binding of charged macromolecules, like polyelectrolytes. The NCD films were grown on p-Si-SiO(2) substrates by microwave plasma-enhanced chemical vapour deposition. To obtain O-terminated surfaces, the NCD films were treated in an oxidising medium. The NCD-based field-effect sensors have been characterised by means of constant-capacitance method. The average pH sensitivity of the O-terminated NCD film was 40 mV/pH. A low detection limit of 5 microM and a high penicillin G sensitivity of 65-70 mV/decade has been obtained for an EDIS penicillin biosensor with the adsorptively immobilised enzyme penicillinase. Alternating potential changes, having tendency to decrease with increasing the number of adsorbed polyelectrolyte layers, have been observed after the layer-by-layer deposition of polyelectrolyte multilayers, using positively charged PAH (poly (allylamine hydrochloride)) and a negatively charged PSS (poly (sodium 4-styrene sulfonate)) as a model system. The response mechanism of the developed EDIS sensors is discussed.
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Técnicas Biossensoriais/instrumentação , Diamante/química , Eletroquímica/instrumentação , Microeletrodos , Nanoestruturas/química , Penicilinase/química , Penicilinas/análise , Transdutores , Técnicas Biossensoriais/métodos , Capacitância Elétrica , Desenho de Equipamento , Análise de Falha de Equipamento , Membranas Artificiais , Nanoestruturas/ultraestrutura , Penicilinas/química , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The light-addressable potentiometric sensor (LAPS) is a semiconductor-based chemical sensor with an electrolyte-insulator-semiconductor structure. The LAPS can have many measuring points integrated on the sensing surface, which are individually accessed by a light beam. By modifying the measuring points with different materials, a single sensor plate can be used as a multi-analyte sensor. In this paper, instrumentation and application of LAPS to multi-ion sensing and imaging are described. As a new application of LAPS, potentiometric imaging of a microfluidic channel is proposed.
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Técnicas Biossensoriais/métodos , Íons/análise , Semicondutores , Técnicas Biossensoriais/instrumentação , Concentração de Íons de Hidrogênio , Luz , Lítio/análise , Potássio/análise , PotenciometriaRESUMO
A new electrochemical detection principle is described for the trace analysis of dissolved species which can be deposited at polycrystalline thin-film metal electrodes and which change the surface resistance of the electrode. Because the latter parameter is measured in dependence on the applied electrode potential this method is called voltohmmetry. The preparation of the required thin-film electrodes and the experimental set-up is introduced and discussed. Typical voltohmmetric experiments are illustrated by measurements of Tl+/Tl at polycrystalline gold electrodes with a thickness of 15 nm. The analytical capabilities of this new approach are discussed. It is already possible to determine heavy metals such as Tl+, Pb2+ or Cd2+ in the range of a few microgram/L by surface resistance-potential measurements at thin-film electrodes with a simple cyclic technique. Further developments of voltohmmetry are envisaged.
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In the present work a double ion sensor based on a laser scanned semiconductor transducer (LSST) for the simultaneous determination of K(+)- and Ca(2+)-ions in solutions has been developed. Specially elaborated ion-sensitive membrane compositions based on valinomycin and calcium ionophore calcium bis[4-(1,1,3,3-tetramethylbutyl)phenyl] phosphate (t-HDOPP-Ca) were deposited as separate layers on a silanized surface of the Si/SiO(2)/Si(3)N(4)-transducer. The proposed multi-sensor exhibits theoretical sensitivities and the detection limits of the sensor were found to be 2 x 10(-6) mol l(-1) for K(+) and 5 x 10(-6) mol l(-1) for Ca(2+). The elaborated double sensor is proposed for the first time as a prototype of a new type of multi-sensor systems for chemical analysis.