摘要:
A self-addressable, self-assembling microelectronic device is designed and fabricated to actively carry out and control multi-step and multiplex molecular biological reactions in microscopic formats. These reactions include nucleic acid hybridizations, antibody/antigen reactions, diagnostics, and biopolymer synthesis. The device can be fabricated using both microlithographic and micro-machining techniques. The device can electronically control the transport and attachment of specific binding entities to specific microlocations. The specific binding entities include molecular biological molecules such as nucleic acids and polypeptides. The device can subsequently control the transport and reaction of analytes or reactants at the addressed specific microlocations. The device is able to concentrate analytes and reactants, remove non-specifically bound molecules, provide stringency control for DNA hybridization reactions, and improve the detection of analytes. The device can be electronically replicated.
摘要:
A method for electronically stabilizing hybridization of nucleic acids bound at a test site of a microelectronic device is described. First and second negatively charged nucleic acids are provided, the second nucleic acid being bound to the test site. A zwitterionic buffer having a conductance of less than 100 mS/cm is applied to the microelectronic device. A current is applied to the test site to positively bias the test site, such that the first negatively charged nucleic acid is transported to the positively biased test site having the bound the second negatively charged nucleic acid. At the test site, the first and second negatively charged nucleic acids hybridize. The zwitterionic buffer acquires a net positive charge under influence of the current, such that the positively charged zwitterionic buffer stabilizes the hybridization by reducing the repulsion between the first and second negatively charged nucleic acids.
摘要翻译:描述了用于电子稳定在微电子器件的测试位点处结合的核酸杂交的方法。 提供第一和第二带负电荷的核酸,第二核酸与测试部位结合。 将具有小于100mS / cm 2的电导的两性离子缓冲液施加到微电子器件。 将电流施加到测试部位以使测试部位正偏置,使得将第一带负电荷的核酸转运到具有结合第二带负电荷核酸的正偏压测试位点。 在测试位点,第一和第二带负电荷的核酸杂交。 两性离子缓冲液在电流影响下获得净正电荷,使得带正电的两性离子缓冲液通过降低第一和第二带负电的核酸之间的排斥来稳定杂交。
摘要:
A self-addressable, self-assembling microelectronic device is designed and fabricated to actively carry out and control multi-step and multiplex molecular biological reactions in microscopic formats. These reactions include nucleic acid hybridizations, antibody/antigen reactions, diagnostics, and biopolymer synthesis. The device can be fabricated using both microlithographic and micro-machining techniques. The device can electronically control the transport and attachment of specific binding entities to specific microlocations. The specific binding entities include molecular biological molecules such as nucleic acids and polypeptides. The device can subsequently control the transport and reaction of analytes or reactants at the addressed specific microlocations. The device is able to concentrate analytes and reactants, remove non-specifically bound molecules, provide stringency control for DNA hybridization reactions, and improve the detection of analytes. The device can be electronically replicated.
摘要:
A system for performing molecular biological diagnosis, analysis and multi-step and multiplex reactions utilizes a self-addressable, self-assembling microelectronic system for actively carrying out controlled reactions in microscopic formats. These reactions include most molecular biological procedures, such as nucleic acid hybridization, antibody/antigen reaction, and clinical diagnostics. Multi-step combinatorial biopolymer synthesis may be performed. A controller interfaces with a user via input/output devices, preferably including a graphical display. Independent electronic control is achieved for the individual microlocations. In the preferred embodiment, the controller interfaces with a power supply and interface, the interface providing selective connection to the microlocations, polarity reversal, and optionally selective potential or current levels to individual electrodes. A system for performing sample preparation, hybridization and detection and data analysis integrates multiple steps within a combined system. Charged materials are transported preferably via free field electrophoresis. DNA complexity reduction is achieved preferably by binding of DNA to a support, followed by cleaving unbound materials, such as by restriction enzymes, followed by transport of the cleaved DNA fragments. Active, programmable matrix devices are formed in a variety of formats, including a square matrix pattern with fanned out electrical connections, an array having electrical connections and optionally optical connections from beneath the specific microlocations. A highly automated DNA diagnostic system results.
摘要:
Apparatus and methods are provided for interacting light with particles, including but not limited to biological matter such as cells, in unique and highly useful ways. Optophoresis consists of subjecting particles to various optical forces, especially optical gradient forces, and more particularly moving optical gradient forces, so as to obtain useful results. In biology, this technology represents a practical approach to probing the inner workings of a living cell, preferably without any dyes, labels or other markers. In one aspect, a particle may be characterized by determining its optophoretic constant or signature. For example, a diseased cell has a different optophoretic constant from a healthy cell, thereby providing information, or the basis for sorting. In the event of physical sorting, various forces may be used for separation, including fluidic forces, such as through the use of laminar flow, or optical forces, or mechanical forces, such as through adhesion. Various techniques for measuring the dielectric constant of particles are provided.
摘要:
Apparatus and methods are provided for interacting light with particles, including but not limited to biological matter such as cells, in unique and highly useful ways. Optophoresis consists of subjecting particles to various optical forces, especially optical gradient forces, and more particularly moving optical gradient forces, so as to obtain useful results. In biology, this technology represents a practical approach to probing the inner workings of a living cell, preferably without any dyes, labels or other markers. In one aspect, a method is provided for separating particles by flowing the particles within a first constrained path, the first constrained path having an input and an output, and a sorting region, the sorting region coupling to a second constrained path, the second constrained path including an output, illuminating the sorting region with a moving optical gradient, characterized in that certain of the particles flow in a laminar manner between the first inlet and the output of the first constrained path, and selected particles are diverted from the first constrained path to the second constrained path under the force of the moving optical gradient.
摘要:
Apparatus and methods are provided for interacting light with particles, including but not limited to biological matter such as cells, in unique and highly useful ways. Optophoresis consists of subjecting particles to various optical forces, especially optical gradient forces, and more particularly moving optical gradient forces, so as to obtain useful results. In biology, this technology represents a practical approach to probing the inner workings of a living cell, preferably without any dyes, labels or other markers. The invention includes methods for separating particles in a medium where the particles having differing dielectric constants by providing a medium having a dielectric constant between the dielectric constants of the particles, subjecting the particles in the media to an optical gradient field, and separating the particles.
摘要:
A method for sorting a particle of interest from a plurality of particles includes the steps of determining an absorption maxima of the particle of interest, providing a light source for generating a beam of coherent light at a wavelength correlating to the absorption maxima, providing a plurality of particles on a support surface, and imparting relative motion between the beam of coherent light and the plurality of particles so as to cause differential movement between the particle of interest and the plurality of particles. The particle of interest is then collected.
摘要:
Apparatus and methods are provided for interacting light with particles, including but not limited to biological matter such as cells, in unique and highly useful ways. Optophoresis consists of subjecting particles to various optical forces, especially optical gradient forces, and more particularly moving optical gradient forces, so as to obtain useful results. In biology, this technology represents a practical approach to probing the inner workings of a living cell, preferably without any dyes, labels or other markers. In one aspect, a method is provided for interacting an optical gradient field in three dimensions with a particle by interfering two beams to generate a plurality of planar fronts, providing a plurality of particles in a medium, and moving the planar fronts relative to the particles, whereby the particles are separated at least in part based upon the dielectric constant of the particles.
摘要:
An electronic device for performing biological operations includes a support substrate and an array of microlocations disposed on the substrate. The array of microlocations include electronically addressable electrodes. A first collection electrode is disposed on the substrate and adjacent to a first side of the array of microlocations. A second collection electrode is disposed on the substrate and adjacent to a second side of the array of microlocations, the second side of the array being opposite of the first side such that the array of microlocations is disposed between the first and second collection electrodes. A flow cell is supported on the substrate. Preferably, the combined area of the collection electrodes is a substantial fraction, preferably at least 50% of the area of the footprint of the flow cell.