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Biomarker Sensors and Method for Multi-Color Imaging and Processing of Single-Molecule Life Signatures

The invention is a device including array of active regions for use in reacting one or more species in at least two of the active regions in a sequential process, e.g., sequential reactions. The device has a transparent substrate member, which has a surface region and a silane material overlying the surface region. A first active region overlies a first portion of the silane material. The first region has a first dimension of less than 1 micron in size and has first molecules capable of binding to the first portion of the silane material. A second active region overlies a second portion of the silane material. The second region has a second dimension of less than 1 micron in size, second molecules capable of binding to the second portion of the active region, and a spatial distance separates the first active region and the second active region.

Wade, Lawrence A.

Selective functionalization of carbon nanotube tips allowing fabrication of new classes of nanoscale sensing and manipulation tools

Embodiments in accordance with the present invention relate to techniques for the growth and attachment of single wall carbon nanotubes (SWNT), facilitating their use as robust and well-characterized tools for AFM imaging and other applications. In accordance with one embodiment, SWNTs attached to an AFM tip can function as a structural scaffold for nanoscale device fabrication on a scanning probe. Such a probe can trigger, with nanometer precision, specific biochemical reactions or conformational changes in biological systems. The consequences of such triggering can be observed in real time by single-molecule fluorescence, electrical, and/or AFM sensing. Specific embodiments in accordance with the present invention utilize sensing and manipulation of individual molecules with carbon nanotubes, coupled with single-molecule fluorescence imaging, to allow observation of spectroscopic signals in response to mechanically induced molecular changes. Biological macromolecules such as proteins or DNA can be attached to nanotubes to create highly specific single-molecule probes for investigations of intermolecular dynamics, for assembling hybrid biological and nanoscale materials, or for developing molecular electronics. In one example, electrical wiring of single redox enzymes to carbon nanotube scanning probes allows observation and electrochemical control over single enzymatic reactions by monitoring fluorescence from a redox-active cofactor or the formation of fluorescent products. Enzymes ''nanowired'' to the tips of carbon nanotubes in accordance with embodiments of the present invention, may enable extremely sensitive probing of biological stimulus-response with high spatial resolution, including product-induced signal transduction.

Wade, Lawrence A.

Biosensors for DNA sequence detection

DNA biosensors are being developed as alternatives to conventional DNA microarrays. These devices couple signal transduction directly to sequence recognition. Some of the most sensitive and functional technologies use fibre optics or electrochemical sensors in combination with DNA hybridization. In a shift from sequence recognition by hybridization, two emerging single-molecule techniques read sequence composition using zero-mode waveguides or electrical impedance in nanoscale pores.

Review

Three-dimensional characterization of tethered microspheres by total internal reflection fluorescence microscopy

Tethered particle microscopy is a powerful tool to study the dynamics of DNA molecules and DNA-protein complexes in single-molecule experiments. We demonstrate that stroboscopic total internal reflection microscopy can be used to characterize the three-dimensional spatiotemporal motion of DNA-tethered particles. By calculating characteristic measures such as symmetry and time constants of the motion, well-formed tethers can be distinguished from defective ones for which the motion is dominated by aberrant surface effects. This improves the reliability of measurements on tether dynamics. For instance, in observations of protein-mediated DNA looping, loop formation is distinguished from adsorption and other nonspecific events.

Non-NASA Center

Fabrication of Buried Nanochannels From Nanowire Patterns

A method of fabricating channels having widths of tens of nanometers in silicon substrates and burying the channels under overlying layers of dielectric materials has been demonstrated. With further refinement, the method might be useful for fabricating nanochannels for manipulation and analysis of large biomolecules at single-molecule resolution. Unlike in prior methods, burying the channels does not involve bonding of flat wafers to the silicon substrates to cover exposed channels in the substrates. Instead, the formation and burying of the channels are accomplished in a more sophisticated process that is less vulnerable to defects in the substrates and less likely to result in clogging of, or leakage from, the channels. In this method, the first step is to establish the channel pattern by forming an array of sacrificial metal nanowires on an SiO2-on-Si substrate. In particular, the wire pattern is made by use of focused-ion-beam (FIB) lithography and a subsequent metallization/lift-off process. The pattern of metal nanowires is then transferred onto the SiO2 layer by reactive-ion etching, which yields sacrificial SiO2 nanowires covered by metal. After removal of the metal covering the SiO2 nanowires, what remains are SiO2 nanowires on an Si substrate. Plasma-enhanced chemical vapor deposition (PECVD) is used to form a layer of a dielectric material over the Si substrate and over the SiO2 wires on the surface of the substrate. FIB milling is then performed to form trenches at both ends of each SiO2 wire. The trenches serve as openings for the entry of chemicals that etch SiO2 much faster than they etch Si. Provided that the nanowires are not so long that the diffusion of the etching chemicals is blocked, the sacrificial SiO2 nanowires become etched out from between the dielectric material and the Si substrate, leaving buried channels. At the time of reporting the information for this article, channels 3 m long, 20 nm deep, and 80 nm wide (see figure) had been fabricated by this method.

Choi, Daniel