Biomedical subjects

S Ingebrandt

Publications and source records attributed to S Ingebrandt.

8 recordsLinked to original sources

PubMed2006

Label-free detection of single nucleotide polymorphisms utilizing the differential transfer function of field-effect transistors.

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.

Biosensing Techniques↗

PubMed2006

Field-effect sensors with charged macromolecules: characterisation by capacitance-voltage, constant-capacitance, impedance spectroscopy and atomic-force microscopy methods.

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.

DNA↗

PubMed2006

Influence of the first amplifier stage in MEA systems on extracellular signal shapes.

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.

Action Potentials↗

PubMed2004

Labelfree fully electronic nucleic acid detection system based on a field-effect transistor device.

The labelfree detection of nucleic acid sequences is one of the modern attempts to develop quick, cheap and miniaturised hand-held devices for the future genetic testing in biotechnology and medical diagnostics. We present an approach to detect the hybridisation of DNA sequences using electrolyte-oxide-semiconductor field-effect transistors (EOSFETs) with micrometer dimensions. These semiconductor devices are sensitive to electrical charge variations that occur at the surface/electrolyte interface, i.e. upon hybridisation of oligonucleotides with complementary single-stranded (ss) oligonucleotides, which are immobilised on the oxide surface of the transistor gate. This method allows direct, time-resolved and in situ detection of specific nucleic acid binding events without any labelling. We focus on the detection mechanism of our sensors by using oppositely charged polyelectrolytes (PAH and PSS) subsequently attached to the transistor structures. Our results indicate that the sensor output is charge sensitive and distance dependent from the gate surface, which pinpoints the need for very defined surface chemistry at the device surface. The hybridisation of natural 19 base-pair sequences has been successfully detected with the sensors. In combination with nano-transistors a PCR free detection system might be feasible in future.

Biosensing Techniques↗

PubMed2003

Backside contacted field effect transistor array for extracellular signal recording.

A new approach to the design of field-effect transistor (FET) sensors and the use of these FETs in detecting extracellular electrophysiological recordings is reported. Backside contacts were engineered by deep reactive ion etching and a gas phase boron doping process of the holes using planar diffusion sources. The metal contacts were designed to fit on top of the bonding pads of a standard industrial 22-pin DIL (dual inline) chip carrier. To minimise contact resistance, the metal backside contacts of the chips were electroless plated with gold. The chips were mounted on top of the bonding pads using a standard flip-chip process and a fineplacer unit previously described. Rat embryonic myocytes were cultured on these new devices (effective growth area 6 x 6 mm(2)) in order to confirm their validity in electrophysiological recording.

Action Potentials↗

PubMed2001

Cardiomyocyte-transistor-hybrids for sensor application.

An extracellular recording system has been designed for the detection of electrical cell signals using p-channel or n-channel field-effect transistor (FET) arrays with non-metallized gates. Signals from rat heart muscle cell were recorded by these devices and the results described on the basis of an equivalent circuit. This technique is sensitive enough to detect minute changes of the extracellular membrane voltage and has potential applications in drug screening. We show that known cardiac stimulants (isoproterenol, norepinephrine) and relaxants (verapamil, carbamylcholine) have characteristic effects on the heart cells in terms of the changes of beat frequencies in the absence or presence of corresponding agents.

Action Potentials↗

PubMed2001

Aligned microcontact printing of biomolecules on microelectronic device surfaces.

Microcontact printing (muCP) of extracellular matrix proteins is a fascinating approach to control cell positioning and outgrowth, which is essential in the development of applications ranging from cellular biosensors to tissue engineering. Microelectronic devices can be used to detect the activity from a large number of recording sites over the long term. However, signals from cells can only be recorded at small sensitive spots. In this paper, we present an innovative setup to perform aligned muCP of extracellular matrix proteins on microelectronic devices in order to guide the growth of electrogenic cells specifically to these sensitive spots. Our system is based on the combination of a fine-placer with redesigned micro stamps having a rigid glass cylinder as backbone for attachment in the alignment tool. Alignment is performed moving the device with an optical table under microscopic control of the superimposed images from stamp and device surface. After successful alignment, the stamp is brought into contact with the device surface by means of a high-precision lever. With our setup, we were able to pattern up to 40 devices per hour. A lateral alignment accuracy of < 2microm has been achieved. Aligned neuronal growth on patterned devices was demonstrated with dissociated hippocampal neurons.

Animals↗

PubMedDate not supplied

Validation of the use of field effect transistors for extracellular signal recording in pharmacological bioassays.

The use of field effect transistors (FETs) in biomedical research has been in rapid progression in recent years. The present study aims to demonstrate a quantitative use of these devices in pharmacological bioassays. FETs were made as a 4 x 4 matrix of gates with a width of 200 microm separating each gate. The surface of the FETs (silicon oxide), covered with a layer of laminin, fibronectin, or nitro cellulose was suitable for cell adhesion. The cultured dissociated cardiac myocytes were spontaneously active within 24 to 48 h after initial plating. Simultaneous intracellular patch clamp recordings were used to verify the electrophysiological signals of cells that were coupled to the gates. All positive chronotropes (isoproterenol, norepinephrine) and negative chronotropes (verapamil, carbamylcholine, SDZ PCO400) showed their characteristic effects on heart cells in terms of changes of beat frequency. As the myocytes were in a complete syncitium on each FET, the cells need not be directly coupled to the gate in order to detect any ionic changes. This enables global cellular responses to be analyzed. The system also offers an opportunity to study the interconnections and communications between different cells. Furthermore, the changes of signal shapes in the presence of different agents could also be detected. The present study demonstrates how versatile and sensitive this recording system is in distinguishing different ionic signal shapes. The authors believe that this system has the potential to replace some currently employed in vitro methods, offering an alternative, which can substantially reduce animal use in pharmacological experiments.

Action Potentials↗