Electrochemical Sensors Based on Enzyme-Linked Immunosorbent Assay

A time-varying electrical field E, having a root-mean-square intensity of 2rms, with a non-zero gradient in a direction transverse to the liquid or fluid flow direction, is produced by a nanostructure electrode array with a very high magnitude gradient near exposed electrode tips. A dielectrophoretic force causes the selected particles to accumulate near the electrode tips, if the medium and selected particles have substantially different dielectric constants. An insulating material surrounds most of the nanostructure electrodes, and a region of the insulating material surface is functionalized to promote attachment of the selected particle species to the surface. An electrical property value Z(meas) is measured at the functionalized surface, and is compared with a reference value Z(ref) to determine if the selected species particles are attached to the functionalized surface. An advantage of this innovation is that an array of nanostructure electrodes can provide an electric field intensity gradient that is one or more orders of magnitude greater than the corresponding gradient provided by a conventional microelectrode arrangement. As a result of the high magnitude field intensity gradients, a nanostructure concentrator can trap particles from high-speed microfluidic flows. This is critical for applications where the entire analysis must be performed in a few minutes.

This invention provides a method and system for fabricating a biomarker sensor array by dispensing one or more entities using a precisely positioned, electrically biased nanoprobe immersed in a buffered fluid over a transparent substrate. Fine patterning of the substrate can be achieved by positioning and selectively biasing the probe in a particular region, changing the pH in a sharp, localized volume of fluid less than 100 nm in diameter, resulting in a selective processing of that region. One example of the implementation of this technique is related to Dip-Pen Nanolithography (DPN), where an Atomic Force Microscope probe can be used as a pen to write protein and DNA Aptamer inks on a transparent substrate functionalized with silane-based self-assembled monolayers. But it would be recognized that the invention has a much broader range of applicability. For example, the invention can be applied to formation of patterns using biological materials, chemical materials, metals, polymers, semiconductors, small molecules, organic and inorganic thins films, or any combination of these.

Because chemical sensors are used in many aspects of space missions, NASA researchers are continually developing ever smaller and more robust sensors that can be manufactured inexpensively and in high quantities; e.g., in batches. Glenn has developed a way to inexpensively fabricate microsensors using a sacrificial template approach. A nanostructure, such as a carbon nanotube, serves as a template, which can then be coated with a high-temperature oxide material. The carbon nanotube can be burned off, or sacrificed, leaving only the metal oxide. The resulting structure provides the unique morphology and properties of the carbon nanotube, which are advantageous for sensing, along with the material durability and high-temperature sensing capabilities of the metal oxide. This technique increases the surface area available for sensing because both the interior and exterior of the resulting microsensor can be used for gas detection, significantly increasing performance. The fabrication of these microsensors includes three major steps: (1) synthesis of the porous metal or metal oxide nanostructures using a sacrificial template, (2) deposition of the electrodes onto alumina substrates, and (3) alignment of the nanostructures between the electrodes. The invention has been demonstrated for methane detection at room temperature (using tin oxide, with carbon nanotubes as the sacrificial template). The microsensor offers low power consumption (no heating required), compact size, extremely low cost, and simple batch-fabrication.

The NASA developed Micro-Organ Device (MOD) platform technology is a small, lightweight, and reproducible in vitro drug screening model that can inexpensively biomimic different mammalian tissues for a multitude of applications. The technology is automated and imposes minimal demands for resources (power, analytes, and fluids). The MOD technology uses titanium tetra(isopropoxide) to bond a microscale support to a substrate and uses biopattering and 3D tissue bioprinting on a microfluidic microchip to eliminate variations in local seeding density while minimizing selection pressure. With the MOD, pharmaceutical companies can test more candidates and concentrate on those with more promise therefore, reducing R&D overall cost. This innovation overcomes major disadvantages of conventional in vitro and in vivo experimentation for purposes of investigating effects of medicines, toxins, and possibly other foreign substances. For example, the MOD platform technology could host life-like miniature assemblies of human cells and the effects observed in tests performed could potentially be extrapolated more readily to humans than could effects observed in conventional in vitro cell cultures, making it possible to reduce or eliminate experimentation on animals. The automated NASA developed technology with minimal footprint and power requirements, micro-volumes of fluids and waste, high throughput and parallel analyses on the same chip, will advance the research and development for new drugs and materials.

Current bulk or thick film solid electrolyte CO2 sensors are expensive, difficult to batch fabricate, and large in size. In contrast, this new amperometric, solid-state, oxide-based electrolyte CO2 microsensor is affordable, easy to fabricate, and is so small that it could easily be integrated onto a substrate the size of a postage stamp. The basic composition of the sensor is identical to a previously designed NASA Glenn technology in which a solid electrolyte of Na3Zr2Si2PO12 is deposited between interdigitated electrodes on an alumina substrate and is covered by Na2CO3/BaCO3. Unlike its predecessor, however, this innovation includes an additional layer of nanocrystalline SnO2 sol gel, an electron donor type (N-type) semiconductor, on top of the Na2CO3/BaCO3 . This new layer provides a greater number of electrons for reduction reaction at the working electrode to detect CO2. As a result, overall performance is enhanced, and this new state-of-the-art sensor has the ability to operate at temperatures as low as 375°C. This low temperature capability significantly decreases the amount of power required to operate the sensor, opening the door to a multitude of new applications that were previously unattainable.