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Biomedical subjects

T Thundat

Publications and source records attributed to T Thundat.

At least 19 recordsLinked to original sources
PubMed2003

Thermal transpiration at the microscale: a Crookes cantilever.

Local temperature inhomogeneities in systems containing micron and submicron objects can result in unexpected consequences. If the mean free path of the host gas constituents is comparable to a characteristic length of the system, then a net exchange of momentum occurs between the constituents and the involved surfaces. For a given temperature gradient and a given pressure range, this results in the presence of Knudsen and Knudsen-like forces (KF). The pressure dependence of these forces has been studied using a microelectromechanical system composed of a microcantilever near a substrate surface. Nano-Newton scale KF are observed by the bending of the microcantilever as monitored by the charge variation on the microcantilever-substrate assembly in a capacitive mode.

Journal Article↗
PubMed2002

Micromechanical measurement of active sites on silicon nitride using surface free energy variation.

We have investigated chemical reactions between adsorbed water and active sites on a silicon nitride surface as a function of temperature and relative humidity using microcantilevers. Effects that might produce a change in the response of the microcantilever, such as a mass adsorption, surface tension of the adsorbed water, and changes in thermal conductivity, were systematically investigated. It is shown that the judicious choice of experimental conditions could make these effects essentially inconsequential in comparison with the instrument response produced by the change in free surface energy of the microcantilever due to the chemical reactions. Using this method, the variation in free surface energy when changing from dry to high humidity conditions was found and the number of active sites that reacted was estimated. This method may be extended to other problems for use in determining surface free energy change and thus the density of reactant sites under different conditions of interest.

Journal Article↗
PubMed2002

An atomic force microscope-based investigation of vertical transport through GaAs/GaAlAs/InAlAs/GaAs step-barrier heterostructures.

Study of vertical transport through heterostructures consisting of single, double, or multiple quantum barriers is of both fundamental and technological interest. While extensive data regarding electron transport is available for single- and double-barrier structures, relatively less information is available for transport through step-barrier structures. In this paper, we present results from a study of room temperature vertical transport through a GaAs/GaAlAs/InAlAs/GaAs multistep-barrier heterostructure. A typical atomic force microscope has been adapted to perform transport measurements, thus allowing precise control of the physical location of the region of measurement. I-V measurements reveal negative differential resistance (NDR) peaks, thus confirming the formation of resonant states in a triangular well created when a voltage bias is applied across the step barrier. I-V curves have also been calculated by numerically solving the Schrödinger wave equation for this step-barrier structure. Comparison between the measured and calculated I-V curves shows reasonable agreement in the number of NDR peaks. However, discrepancies exist between the measured and calculated values for the voltages at which these NDR peaks occur. Some possible reasons for these discrepancies are discussed in this work.

Journal Article↗
PubMed2001

Cantilever-based optical deflection assay for discrimination of DNA single-nucleotide mismatches.

Characterization of single-nucleotide polymorphisms is a major focus of current genomics research. We demonstrate the discrimination of DNA mismatches using an elegantly simple microcantilever-based optical deflection assay, without the need for external labeling. Gold-coated silicon AFM cantilevers were functionalized with thiolated 20- or 25-mer probe DNA oligonucleotides and exposed to target oligonucleotides of varying sequence in static and flow conditions. Hybridization of 10-mer complementary target oligonucleotides resulted in net positive deflection, while hybridization with targets containing one or two internal mismatches resulted in net negative deflection. Mismatched targets produced a stable and measurable signal when only a four-base pair stretch was complementary to the probe sequence. This technique is readily adaptable to a high-throughput array format and provides a distinct positive/negative signal for easy interpretation of oligonucleotide hybridization.

Base Pair Mismatch↗
PubMed2001

Ultrasensitive detection of CrO4(2-) using a microcantilever sensor.

Microcantilevers modified with a self-assembled monolayer respond sensitively to specific ion concentrations. Here, we report the detection of trace amounts of CrO4(2-) using microcantilevers modified with a self-assembled monolayer of triethyl-12-mercaptododecylammonium bromide. The self-assembled monolayer was prepared on a silicon microcantilever coated with a thin layer of gold on one side. The microcantilever undergoes bending due to sorption of CrO4(2-) ions on the monolayer-modified side. It was found that a concentration of 10(-9) M CrO4(2-) can be detected using this technology in a flow cell. Other anions, such as Cl-, Br-, CO3(2-) (or HCO3-), and SO4(2-), have minimal effect on the deflection of this cantilever. The mechanics of the bending and the chemistry of cantilever modification are discussed.

Journal Article↗
PubMed2001

Origin of nanomechanical cantilever motion generated from biomolecular interactions.

Generation of nanomechanical cantilever motion from biomolecular interactions can have wide applications, ranging from high-throughput biomolecular detection to bioactuation. Although it has been suggested that such motion is caused by changes in surface stress of a cantilever beam, the origin of the surface-stress change has so far not been elucidated. By using DNA hybridization experiments, we show that the origin of motion lies in the interplay between changes in configurational entropy and intermolecular energetics induced by specific biomolecular interactions. By controlling entropy change during DNA hybridization, the direction of cantilever motion can be manipulated. These thermodynamic principles were also used to explain the origin of motion generated from protein-ligand binding.

DNA, Single-Stranded↗
PubMed2001

Measuring magnetic susceptibilities of nanogram quantities of materials using microcantilevers.

We describe a novel technique for measuring magnetic susceptibilities of nanogram quantities of magnetic materials that utilizes the extreme force sensitivity of microcantilevers. The magnetic force acting on samples attached to the free end of a cantilever can be measured as changes in the resonance response of the cantilever. The shift in resonance frequency of the cantilever is proportional to the field gradient, whereas the deflection of a cantilever is proportional to the magnetic force. The magnetic susceptibility measurement is based on comparison of the forces acting on the sample and a reference material in the same magnetic field and field gradient. We have determined the magnetic susceptibilities of nanogram quantities of many paramagnetic materials. The measured magnetic susceptibilities show excellent agreement with values found in the literature.

Journal Article↗
PubMed2001

Bioassay of prostate-specific antigen (PSA) using microcantilevers.

Diagnosis and monitoring of complex diseases such as cancer require quantitative detection of multiple proteins. Recent work has shown that when specific biomolecular binding occurs on one surface of a microcantilever beam, intermolecular nanomechanics bend the cantilever, which can be optically detected. Although this label-free technique readily lends itself to formation of microcantilever arrays, what has remained unclear is the technologically critical issue of whether it is sufficiently specific and sensitive to detect disease-related proteins at clinically relevant conditions and concentrations. As an example, we report here that microcantilevers of different geometries have been used to detect two forms of prostate-specific antigen (PSA) over a wide range of concentrations from 0.2 ng/ml to 60 microg/ml in a background of human serum albumin (HSA) and human plasminogen (HP) at 1 mg/ml, making this a clinically relevant diagnostic technique for prostate cancer. Because cantilever motion originates from the free-energy change induced by specific biomolecular binding, this technique may offer a common platform for high-throughput label-free analysis of protein-protein binding, DNA hybridization, and DNA-protein interactions, as well as drug discovery.

Biosensing Techniques↗
PubMed1997

Mapping individual cosmid DNAs by direct AFM imaging.

Individual cosmid clones have been restriction mapped by directly imaging, with the atomic force microscope (AFM), a mutant EcoRI endonuclease site-specifically bound to DNA. Images and data are presented that locate six restriction sites, predicted from gel electrophoresis, on a 35-kb cosmid isolated from mouse chromosome 7. Measured distances between endonuclease molecules bound to lambda DNA, when compared to known values, demonstrate the accuracy of AFM mapping to better than 1%. These results may be extended to identify other important site-specific protein-DNA interactions, such as transcription factor and mismatch repair enzyme binding, difficult to resolve by current techniques.

Animals↗
PubMed1996

Direct atomic force microscope imaging of EcoRI endonuclease site specifically bound to plasmid DNA molecules.

Direct imaging with the atomic force microscope has been used to identify specific nucleotide sequences in plasmid DNA molecules. This was accomplished using EcoRI (Gln-111), a mutant of the restriction enzyme that has a 1000-fold greater binding affinity than the wild-type enzyme but with cleavage rate constants reduced by a factor of 10(4). ScaI-linearized plasmids with single (pBS+) and double (pGEM-luc and pSV-beta-galactosidase) EcoRI recognition sites were imaged, and the bound enzyme was localized to a 50- to 100-nt resolution. The high affinity for the EcoRI binding site exhibited by this mutant endonuclease, coupled with an observed low level of nonspecific binding, should prove valuable for physically mapping large DNA clones by direct atomic force microscope imaging.

Binding Sites↗
PubMed1994

Stretched DNA structures observed with atomic force microscopy.

Double-stranded DNA molecules are occasionally found that appear to be straightened and stretched in atomic force microscope (AFM) images. Usually pBS+ plasmid and lambda DNA show relaxed structures with bends and kinks along the strands and have measured contour lengths consistent to about 5-7%; they also appear not to cross over each other, except in very high concentrations. The anomalous molecules observed here, compared with the majority of molecules in the preparation, show contour lengths increased by as much as 80% and have measured heights of about half that of normal relaxed DNA. Some molecules also appear to be in transition between stretched and relaxed forms. These observations are consistent with an uncoiling of the DNA helix without breakage of the covalent bonds in the deoxyribose-phosphate backbone.

Bacteriophage lambda↗
PubMed1992

Immobilization of DNA for scanning probe microscopy.

Reproducible scanning tunneling microscope and atomic force microscope images of entire molecules of uncoated plasmid DNA chemically bound to surfaces are presented. The chemically mediated immobilization of DNA to surfaces and subsequent scanning tunneling microscope imaging of DNA molecules demonstrate that the problem of molecular instability to forces exerted by the probe tip, inherent with scanning probe microscopes, can be prevented.

DNA, Bacterial↗
PubMed1992

Electrostatic spraying of DNA molecules for investigation by scanning tunneling microscopy.

We have investigated electrostatic spraying of DNA onto gold surfaces as an alternative sample-preparation technique for STM studies. Preliminary results show that a higher distribution of isolated strands as well as well ordered aggregates can be obtained with this technique when compared with electrodeposition or drop evaporation. In many places, the well ordered aggregates were found to cleave in a direction perpendicular to their length after repeated scanning in the same direction.

DNA↗
PubMed1992

Scanning tunneling microscopy of DNA: a novel technique using radiolabeled DNA to evaluate chemically mediated attachment of DNA to surfaces.

pBS+ plasmid deoxyribonucleic acid (DNA) was imaged by scanning tunneling microscopy (STM) after mounting microdroplets by aerosol deposition onto heated epitaxial gold surfaces. However, the instability of the adsorbate to forces exerted by the tunneling tip points out the need for more aggressive bonding of molecules to surfaces. We describe a sensitive assay for the qualitative and quantitative evaluation of chemical agents to influence binding of DNA to surfaces using 32P-labeled pBS+ plasmid DNA. We propose that such an assay can make an important contribution to immobilization techniques prior to STM imaging.

DNA↗