Abstract: Probe sets for the multiplexed detection of the binding of, or interaction between, one or more ligands and target antiligands are provided. Detection involves the release of identifying tags as a consequence of target recognition. The probe sets include electrophoretic tag probes or e-tag probes, comprising a detection region and a mobility-defining region called the mobility modifier, both linked to a target-binding moiety. The probes comprise interactive functionalities adjacent the cleaved portion positioned in the probes such that the interactive functionality does not form part of the e-tag reporters. Target antiligands are contacted with a set of e-tag probes and the contacted antiligands are treated with a selected cleaving agent resulting in a mixture of e-tag reporters and uncleaved and/or partially cleaved e-tag probes. The mixture is exposed to a capture agent effective to bind to uncleaved or partially cleaved e-tag probes, followed by electrophoretic separation.

Abstract: Kits for the multiplexed detection of the binding of, or interaction between, one or more ligands and target antiligands are provided. Detection involves the release of identifying tags as a consequence of target recognition. The kits include sets of electrophoretic tag probes or e-tag probes, a capture agent and optionally a cleaving agent. The e-tag probes comprise a detection region and a mobility-defining region called the mobility modifier, both linked to a target-binding moiety. In using the kits, target antiligands are contacted with a set of e-tag probes and the contacted antiligands are treated with a selected cleaving agent resulting in a mixture of e-tag reporters and uncleaved and/or partially cleaved e-tag probes. The mixture is exposed to a capture agent effective to bind to uncleaved or partially cleaved e-tag probes, followed by electrophoretic separation. In a multiplexed assay, different released e-tag reporters may be separated and detected providing for target identification.

Abstract: Devices and methods are disclosed for moving charged molecules through a medium by the application of a plurality of electrical fields of sufficient strength and applied for sufficient amounts of time so as to move the charged molecules through the medium. The devices although preferably small in size, preferably generate large numbers (100 or more) of electrical fields to a movement area which preferably contains a liquid buffered or gel medium. Mixtures of charged molecules are pulled through the gel by the force of the electrical fields. The fields are preferably activated simultaneously or sequentially one after another at various speeds to create complex force field distributions or moving field waves along the separation medium. Charged molecules capable of moving quickly through the gel will be moved along by the faster moving field waves and be separated from slower moving molecules.

Abstract: Kits for the multiplexed detection of known, selected nucleotide target sequences are provided. Detection involves the release of identifying tags as a consequence of target recognition. The kits include sets of electrophoretic tag probes or e-tag probes, capture agent and optionally a nuclease. The e-tag probes comprise a detection region and a mobility-defining region called the mobility modifier, both linked to a target-binding moiety. In using the kits, the target-binding moiety of the e-tag probes hybridizes to complementary target sequences followed by nuclease cleavage of the e-tag probes and release of detectable e-tags or e-tag reporters. The mixture is exposed to a capture agent which binds uncleaved and/or partially cleaved e-tag probes, followed by electrophoretic separation. In a multiplexed assay, different released e-tag reporters may be separated and detected providing for target identification.

Abstract: The present invention is directed to methods and compositions for determining the presence, absence, and/or amounts of one or more membrane-associated analytes in a sample. In accordance with the invention, binding compounds derivatized with releasable molecular tags specifically bind to selected membrane-associated analytes, after which the molecular tags are released upon activation of cleavage moieties, or sensitizers, anchored in the same membrane as the membrane-associated analytes. The released molecular tags are then identified by their distinct separation and detection characteristics.

Abstract: A microfluidic device having integrated components for conducting chemical operations. Depending upon the desired application, the components include electrodes for manipulating charged entities, heaters, electrochemical detectors, sensors for temperature, pH, fluid flow, and other useful components. The device may be fabricated from a plastic substrate such as, for example, a substantially saturated norbornene based polymer. The components are integrated into the device by adhering an electrically conductive film to the substrate. The film may be made of metal or an electrically conducting ink and is applied to the device through metal deposition, printing, or other methods for applying films. Methods for reducing bubble formation during electrokinetic separation and methods for heating material in a microfluidic device are also disclosed.

Abstract: Probe sets for the multiplexed detection of known, selected nucleotide target sequences are provided. Detection involves the release of identifying tags as a consequence of target recognition. The probe sets include electrophoretic tag probes or “e-tag probes”, comprising a detection region and a mobility-defining region called the mobility modifier, both linked to a target-binding moiety. The target-binding moiety of the e-tag probes hybridizes to complementary target sequences followed by nuclease cleavage of the e-tag probes and release of detectable e-tags or e-tag reporters. The mixture is exposed to a capture agent which binds uncleaved and/or partially cleaved e-tag probes, followed by electrophoretic separation. In a multiplexed assay, different released e-tag reporters may be separated and detected providing for target identification.

Abstract: Improved sealing for microstructures in microfluidic devices having a plurality of units is provided by providing collars surrounding the openings to the microstructures, such as reservoirs. The collars are protrusions extending from the surface of the devices and the internal walls of the collars generally aligned with the internal walls of the microstructure. Conformable and/or adhesive lids are employed for sealing the microstructures.

Abstract: A method for performing an electrophoretic separation on a microfluidic device includes applying an electric field to electrokinetically move a sample along a channel towards a location. The sample may be concentrated by application of the electric field. The method further comprises optically monitoring the location for at least a portion of the sample. Once the sample is detected, the electric field is automatically changed to further manipulate the sample. The sample may be spatially separated along a separation channel or channel portion. The closed-loop control of electric fields may be performed using a control unit adapted to apply a voltage potential between electrodes and an optical detector such as an LIF detector.

Abstract: Devices are provided comprising reflecting channels for directing horizontally a vertical light beam through a multiplicity of streams. The devices provide particular applications in microfluidics, where streams of narrow cross-sectional dimensions are involved. Desirably, channels having 45° walls are provided for directing the beams and where the streams are divided by walls, the walls are at substantially 90°. Methods of fabrication of the devices are provided.

Abstract: A system for material excitation in microfluidic devices is described. Aspects of the invention resemble a submerged periscope when in use. They allow for light to be redirected along a more advantageous trajectory as does the maritime device. A prism with a reflecting surface may be used to direct a laser beam along one or more microfluidic trenches or channels. Alternatively, a reflecting surface may be provided in connection with a simple support. Directing a beam along multiple paths is preferably accomplished by scanning a single laser across or around the reflecting surface provided. Provision may be made for at least a portion of the submersible used to function as an electrode to assist in electrokinetically driving fluids and/or ions within the microfluidic device.

Abstract: A mutiplexed enzyme assay method and substrate set for performing the method are disclosed. The method includes performing a plurality of enzyme reactions in the presence of a plurality of enzymes substrates, under conditions effective to convert an enzyme substrate to a corresponding product, where the product of each substrate and the substrate have different separation characteristics from each other and from the other substrates and their corresponding products. After performing the reactions, which may be carried out in separate or combined reactions, the substrates and products in said reactions are separated in a single separation medium. For each separated product and substrate, a separation characteristic effective to identify that product and substrate and a signal related to the amount of the product and substrate is detected. From this, one can determine the amount of substrate converted to the corresponding product in each of the reactions.

Abstract: The invention relates to a method for linking two adjacent plastic work pieces (2, 3). The inventive method is characterized in that the intended contact zone (K) of at least one of the two work pieces (2) with which the latter adjoins to the other work piece (3) is at least partially subjected to a high-energy radiation to such an extent that the glass transition temperature is reduced in a marginal zone (R). The two work pieces (2, 3) are brought in the desired position relative to each other. For establishing the connection of the two work pieces (2, 3) at least the marginal zone (R) modified by the previous radiation process is heated at least in its surface area to a temperature that is higher than the glass transition temperature of the marginal zone (R) modified by the radiation process but lower than the glass transition temperature of the zones of the work pieces (2, 3) that have not been modified.

Abstract: An optical detection and orientation device is provided comprising a housing having an excitation light source, an optical element for reflecting the excitation light to an aspherical lens and transmitting light emitted by a fluorophore excited by said excitation light, a focusing lens for focusing the emitted light onto the entry of an optical fiber, which serves as a confocal aperture, and a moveable carrier for accurately moving said housing over a small area in relation to a channel in a microfluidic device. The optical detection and orientation device finds use in detecting fluorophores in the channel during operations involving fluorescent signals.

Abstract: A continuous form microstructure array device is constructed as a flexible elongate film laminate containing a plurality of microstructures or arrays of microstructures arranged serially lengthwise along the laminate. The laminate can be continuously drawn from a supply roll or stack, advanced within an analytic device and, when analysis is complete, taken up in a storage roll or stack. The device provides for high throughput microfluidic processing.

Abstract: Microfluidic devices are provided having units of 8-fold symmetry comprising 8 assay units, where a reservoir provides a common component to 8 assay units. The units can be compactly formed in a substrate to provide the ability to perform a large number of assays within a small area. The microfluidic devices find use in operations, such as assays, DNA sequence detection, etc. Various formats can be used to have the microfluidic device interrelate with microtiter well plates. Methods are provided for monitoring the flow rates/velocities of assay components and streams for comparison of results in different assay units and/or to modify the conditions to change the flow rates in particular channels.

Abstract: A method and device for injecting a liquid sample into an electrolyte channel in a microfluidics device is disclosed. The device has a channel network that includes an electrolyte channel having upstream and downstream channel portions and first, second, and third side channels that intersect the electrolyte channel between the two channel portions at first, second, and third ports, respectively. In the method, a sample is moved electrokinetically into the electrolyte channel, to form a defined sample volume therein. By simultaneously controlling the voltage applied to the three side channels, and at least one of the upstream and downstream channel end portions, the sample volume element can be shaped to have a desired leading- and trailing-edge shape and/or distribution of sample components within the volume elements.

Abstract: Methods for the multiplexed detection of the binding of, or interaction between, one or more ligands and target antiligands are provided. Detection involves the release of identifying tags as a consequence of target recognition. The methods include the use of electrophoretic tag probes or e-tag probes, comprising a detection region and a mobility-defining region called the mobility modifier, both linked to a target-binding moiety. In practicing the methods, target antiligands are contacted with a set of e-tag probes and the contacted antiligands are treated with a selected cleaving agent resulting in a mixture of e-tag reporters and uncleaved and/or partially cleaved e-tag probes. The mixture is exposed to a capture agent effective to bind to uncleaved or partially cleaved e-tag probes, followed by electrophoretic separation. In a multiplexed assay, different released e-tag reporters may be separated and detected providing for target identification.

Abstract: Devices and methods are disclosed for moving charged molecules through a medium by the application of a plurality of electrical fields of sufficient strength and applied for sufficient amounts of time so as to move the charged molecules through the medium. The devices although preferably small in size, preferably generate large numbers (100 or more) of electrical fields to a movement area which preferably contains a liquid buffered or gel medium. Mixtures of charged molecules are pulled through the gel by the force of the electrical fields. The fields are preferably activated simultaneously or sequentially one after another at various speeds to create complex force field distributions or moving field waves along the separation medium. Charged molecules capable of moving quickly through the gel will be moved along by the faster moving field waves and be separated from slower moving molecules.

Abstract: A continuous form microstructure array device (20) is constructed as a flexible elongate film laminate containing microstructure arrays (26) arranged serially along the laminate. The laminate can be continuously drawn from a roll, passed through a processing and analysis device and rerolled or stacked for storage.