Abstract: A method and apparatus for correcting an image of an electrophoresis gel for lens and detector non-uniformities is provided. The removal of such non-uniformities, in conjunction with a uniform illumination source, allows quantitative measurements of an electrophoresis gel to be made, thus increasing the information which can be obtained from an electrophoretic analysis. Lens and detector non-uniformities are removed by first determining the magnitude of the non-uniformities using a calibration standard. The calibration standard has a uniform emittance, preferably in the same wavelength band as the labeled regions of the electrophoresis gel. By placing the calibration standard in close proximity to the entrance aperture of the lens, the calibration process is relatively insensitive to illumination source non-uniformities. Thus an image of the calibration standard taken with the same lens settings as the unknown provides detailed information on the lens and detector non-uniformities.

Abstract: Methods and apparatus are disclosed for improved mapping and sequencing of carbohydrate polymers. In one aspect, the improvement includes a means for coupling two capillary electrophoresis (CE) tubes in such a way so as to (i) efficiently transfer a selected sample component from a first CE capillary to a second CE capillary or, (ii) introduce a supplementary reagent into the separation path between the two capillaries, e.g., an internal standard, binding agent, enzyme, and the like. In an additional aspect, the invention includes improved apparatus and methods for electrophoresis of labeled carbohydrates in which a sample carbohydrate labeled with a first label is separated by CE and its migration behavior is compared with an internal standard labeled with one or more second labels which are distinguishable from the first label.

Abstract: The present invention provides a method for direct detection concurrently with separation in capillary electrophoresis using a lanthanide cation selected from the group consisting of Tb(III), Eu(III), Dy(III), and Sm(III) to provide fluorescence for enhanced sensitivity. Analytes separated and concurrently detected using this method have an absorption spectrum from about 230 nm to 400 nm and include compounds having an .alpha.,.beta. unsaturated carbonyl group, an .alpha.-keto acid, an aromatic moiety, indole carboxylic acid, a diketone moiety, or an organic acid, for example, a salicylic acid, a carboxylic acid, or an amino acid. Advantages of this method include the lack of derivatization, either pre- or post-column, loss of analyte during derivatization is avoided, and fewer items are needed for a commercial capillary electrophoresis kit.

Abstract: The present invention system includes an axially oriented end of a fiber optic present in an axially oriented system component bore in which, during use, sample analyte fluorescence is caused to occur. The present invention system and method provides that sample analyte(s) fluorescence inducing energy be entered along a path which is other than essentially parallel to the axially oriented system component bore and that detected fluorescence be transmitted to a detector by the fiber optic. A preferred source of sample analyte(s) fluorescence inducing energy is a laser system, and a preferred method by which to provide sample analyte(s) to the present invention system axially oriented system component bore involves use of electrophoresis.

Abstract: An electrophoresis apparatus includes a plurality of first capillaries containing a separation medium, such as a gel, and an optical cell in which one end of each of the first capillaries is disposed. Samples labeled with fluorophores and introduced into the first capillaries, and an electric field is applied to the first capillaries to cause the samples to migrate through the first capillaries into the optical cell. A light source excites the fluorophores with light when the samples are in the optical cell, causing the fluorophores to emit fluorescence, and a photo-detecting system detects the fluorescence emitted by the fluorophores. The ends of the first capillaries in the optical cell may be arranged in a straight line. The light source may include a He--Ne laser emitting light having a wavelength of 594 nm, and the fluorophores may include either sulforhodamine or a derivative of sulforhodamine.

Abstract: A system for processing a plurality of tests or syntheses in parallel comprising a sample channel for moving samples into a microlaboratory array of a plurality of wells connected by one or more channels for the testing or synthesis of samples, a station for housing the array and an optical system comprising at least one light source and at least one light detector for measuring the samples in the array, and a means of electrically connecting said array to an apparatus capable of monitoring and controlling the flow of fluids into the array.Samples are loaded from a common loading channel into the array, processed in the wells and measurements taken by the optical system. The array can process many samples, or synthesize many compounds in parallel, reducing the time required for such processes.

Abstract: A device for combined bioaffinity assay and electrophoretic separation is provided, which comprises a capillary system having two stages, a first stage in which bioaffinity interactions of analyte molecules and molecular recognition elements are performed, and a second stage, in which electrophoretic separation of the analyte molecules and subsequent detection of the separated species is accomplished. Within the first capillary stage the molecular recognition elements are attached and immobilized to the inside capillary wall, for example, by adsorption or by covalent binding to the capillary material. The method for combined bioaffinity assay and electrophoretic separation comprises flowing an analyte through a capillary system having two stages. In a first capillary stage the analyte molecules are captured by respective molecular recognition elements present in that stage.

Abstract: The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

Abstract: The visualization of RNA bands in denaturing electrophoresis gels is improved by staining the gels with a 2,7-diamino-10-(N,N,N',N'-tetramethyl-1,3-propanediamino)-propyl-9-phenylp henanthridine dihalide hydrohalide or a 2,7-diamino-10-?3-(diethylmethylammonio)propyl!-9-phenylphenanthridine dihalide. These dyes provide effective contrast for sensitive detection without the need for extensive washing of excess dye from the gels to lower background noise.

Abstract: A capillary array electrophoresis system by which measurement is conducted using a large number of capillaries. The electrophoresis system includes a plurality of capillary array sheets stacked one on top of another wherein end portions of the capillaries at a detection region are arranged two-dimensionally in such a manner as to elute two-dimensionally a sample from the distal end of each capillary. Excitation light is applied to the sample eluted into a buffer solution, and a two-dimensional fluorescent image is picked up by a detector.

Abstract: To detect bands in an electrophoresis capillary tube having a liquid separating medium, a light source and a light detector of a monitor are positioned on opposite sides of the capillary tube with the light source transmitting light through a first slit and light passing to the detector through a second slit after it has passed through the capillary tube. The first and second slits are aligned with the direction of motion of bands, have a maximum length of less than 500 micrometers and a maximum width of less than 200 micrometers. There are no vertical lengths in the capillary tube having a dimension greater than one third of its length between a sample injecting end and a detecting end of the capillary tube.

Abstract: A first reactant is immobilized i.e. in a porous matrix (50), adjacent a sample electrode (46) within a reaction chamber. Energizing of the electrode (46) electrophoretically attracts a mobile second reactant and/or electrolytically induces appropriate reaction conditions to enhance reaction of the first and second reactants. Polarity reversals between the sample electrode (46) and remote electrodes (38), (42), (44) cause unreacted second reactant and/or by-products to migrate away from the immobilized first reactant. The techniques are useful for sequential chemical reactions such as sequencing or construction of proteins, polysaccharides and nucleic acids where cyclical additions and removals of reactants are required. The techniques are amenable to automated micro and nano scale construction and operation and allow direct electrophoretic (38) interfacing with chromatographic, HPCE and mass spectrophotometric equipment.

Abstract: A process for detecting a target nucleic acid, which includes the steps of hybridizing a target single-stranded nucleic acid in a sample solution to a labeled single-stranded nucleic acid probe to form a reaction mixture containing a hybrid, subjecting the reaction mixture to capillary electrophoresis to separate the hybrid from the free labeled nucleic acid probe, and detecting the hybrid utilizing the label moiety of the nucleic acid probe constituting the hybrid.

Abstract: A technique and circuit for measuring direct current flowing through a conductor at a high voltage employs a capacitor connected in series with the conductor, and a neon lamp connected in parallel with the capacitor. The series connected capacitor and conductor are connected to a high voltage source such that the current flowing through the series connected capacitor and conductor charges up the capacitor. Since the capacitor and the neon lamp are connected in parallel, the voltage across the electrodes of the neon lamp follows the voltage across the capacitor. When the voltage across the electrodes of the neon lamp reaches the lamp's ignition voltage, the neon lamp fires, and discharges the capacitor until the voltage across the electrodes of the neon lamp falls to the lamp's extinction voltage. The light signal generated by the neon lamp is picked up by a fiber optic cable and transmitted to a remote receiver for processing.

Abstract: Compositions, methods, and apparatus for performing ultrafast binding assays by capillary electrophoresis or other electroseparation techniques are disclosed. In one embodiment, a first binding partner carries a detectable label and a second binding partner is modified to be highly charged. When used in combination with a sample containing an analyte with which both binding partners can interact and bind thereto, a three-membered complex is formed. The electrophoretic mobility difference between the unbound and complex-bound forms of labeled first binding partner is such that electroseparation and subsequent detection of an analyte can be accomplished. The compositions, methods, and apparatus disclosed herein also permit quantitative determination of the concentration of an analyte in a sample.

Abstract: The present invention comprises a method Of detecting and quantifying components separated by capillary electrophoresis or other techniques using thin capillaries in the separation. The method of the present invention comprises irradiating the whole or large parts of the capillary at short time intervals and detecting the fluorescence originating from the sample components, preferably by a CCD detector (charge coupled device). Thereby, an instantaneous image of the progress of the separation process is obtained. The present invention further comprises an apparatus for performing this detection.

Abstract: The electrophoresis apparatus has a capillary filled with gel for use in electrophoresis. A first buffer solution container is provided for storing a buffer solution and introducing a sample labelled with a fluorescent substance into an inlet of said capillary for electrophoresis. A second buffer solution container is provided for storing a buffer solution containing a luminous solution, into which said sample is continually introduced from an outlet of said capillary after electrophoresis. Electrophoresis means is provided for subjecting said sample to electrophoresis by applying a predetermined value of voltage to said gel through which said sample is being transferred while being electrophoresed. Light receiving means is disposed underneath the outlet of said capillary to read fluorescence emitted from said sample from bottom in a direction in which said sample is approaching closer.

Abstract: A system and method are disclosed for optically aligning a capillary tube and an excitation laser beam for fluorescence detection applications by utilizing the Raman scatter signals of the capillary tube's contents. For example, Raman scatter by an electrophoretic separation matrix may be used for alignment in a capillary electrophoresis system. Fluorescent material may be present and may also be used for alignment purposes, but is not necessary. The invention employs a parabolic reflector, having apertures through which the capillary tube and the laser beam are guided so that they intersect, preferably at right angles and at the focal point of the reflector. The Raman scatter signals of the material within the capillary tube are collected via a series of filters and this information is used to reposition, if necessary, a focusing lens that directs the excitation beam into the reflector and the capillary tube, so that the Raman scatter signals are maximized.

Abstract: Disclosed is a method for and an apparatus for detecing amines or amino acids, which apparatus generally includes a capillary electrophoresis separation tube with a post-capillary reactor positioned at the end of the tube to immediately receive separated samples from the tube. The post-capillary reactor includes a solution of Ru(bpy).sub.3.sup.2+ buffered with a base. The post capillary reactor further includes an electrode assembly for providing current to the solution to convert nonluminescing Ru(bpy).sub.3.sup.2+ to luminescing Ru(bpy).sub.3.sup.3+. The method generally includes separating the desired analyte from the sample, contacting the analyte with the solution to produce luminescence, and then photometrically measuring the amount of analyte present as a function of the luminescence.

Abstract: An assembly suitable for identifying a code sequence of at least a portion of a biomolecule in a free-solution embodiment. The assembly comprises first means for migrating and separating a portion of a biomolecule in a free-solution; second means comprising a near-field probe for generating a super-resolution chemical analysis of a portion of a biomolecule; and, third means for correlating the super-resolution chemical analysis of the portion of the biomolecule with a broad spectral content of a referent biomolecule, for generating a code sequencing of the portion of the biomolecule.

Abstract: A capillary and capillary retaining system including a capillary assembly having first and second end holders. The first and second end holders are adapted to be received by first and second retainers. The first end holder may include protruding portions and the first end retainer includes clips or clamps to engage the protruding portions. The second end holder may include opposite recesses and the second retainer is adapted to receive optical cables that are received within the recesses to retain the second end holder. The second retainer can further include a lever on the retainer body to pry the second holder out of the second retainer. Locks or clips retained by the second end retainer cooperate with grooves in the optical cables to retain the optical cables.

Abstract: A buffer and method useful for the analysis of glycoproteins by capillary zone electrophoresis. The buffer comprises water, a sugar complexing compound, a base compound for adjusting the pH, and a zwitterionic compound. An embodiment of the buffer comprises sodium borate as the complexing compound, sodium hydroxide as the base, and 3-cyclohexylamino-1-propanesulfonic acid as the zwitterionic compound. In the method, a selected glycoprotein is subjected to capillary zone electrophoresis. The proportion or the amount of the glycoprotein is determined by quantitative analysis of the resulting electropherogram.

Abstract: A system for interfacing Capillary Zone Electrophoresis (CZE) systems to Inductively Coupled Plasma-Mass Spectrometer (ICP-MS) sample analysis systems is disclosed. The interface system includes a Sample Introduction Electrophoresis Capillary Tube as an integral part of a modified Direct Injection Nebulizer (DIN) system. During use sample components separated by (CZE) are entered to the modified (DIN) via a sample introduction electrophoresis capillary tube present therein, in which modified (DIN) they are mixed with an entered make-up liquid. The make-up liquid is conductive and serves to make electrophoresis effecting electrical contact to the end of the sample introduction electrophoresis capillary tube in the modified (DIN).

Abstract: A multiple capillary analyzer allows detection of light from multiple capillaries with a reduced number of interfaces through which light must pass in detecting light emitted from a sample being analyzed, using a modified sheath flow cuvette. A linear or rectangular array of capillaries is introduced into a rectangular flow chamber. Sheath fluid draws individual sample streams through the cuvette. The capillaries are closely and evenly spaced and held by a transparent retainer in a fixed position in relation to an optical detection system. Collimated sample excitation radiation is applied simultaneously across the ends of the capillaries in the retainer. Light emitted from the excited sample is detected by the optical detection system. The retainer is provided by a transparent chamber having inward slanting end walls. The capillaries are wedged into the chamber.

Abstract: The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

Abstract: An apparatus for collecting samples from capillary electrophoresis (CE) for matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS) is disclosed. The apparatus has a capillary, a metallic support, a porous member, and a power supply for applying an electrical potential between the porous member and an inlet end of the capillary to drive the sample through the capillary during CE. The capillary transmits a sample from the inlet end to the exit end during capillary electrophoresis. The metallic support supports the porous membrane such that the porous membrane contacts the capillary exit end during CE. The metallic support (with the porous membrane) is suitable for placing in a mass spectrometer to act as a repeller for MALDI. The porous member has at least a portion thereof that is generally concentric with the capillary exit end and contacts the porous membrane during CE.

Abstract: A substrate defines a group therein having three sections: a first and third sections having dimensions larger than the outside dimensions of two capillaries and a second section separating the first and third sections with dimensions small than the outside dimensions of the capillaries. When the two capillaries are inserted into the first and third sections and capillary electrophoretic separation performed in one or both of the capillaries, the separated components may be detected using a pair of detectors located at the second section. The electrodes are electrically isolated from an analysis or storage device used to analyze or store signals at the electrodes.

Abstract: Homogeneous immunoassays and enzyme-substrate assays which use capillary electrophoresis and fluoescent detection systems are described. The method is useful for detecting and/or quantitating the concentration of analytes in a sample.

Abstract: Processes for the electrophoretic analysis of monosaccharides and/or polysaccharides are disclosed. The processes generally first include providing a composition having derivatized mono and/or polysaccharide components which are monosaccharide and/or polysaccharide labelled with a fluorescing charged compound. Then introducing the composition into an electrophoretic medium and applying an electric field across the electrophoretic medium causes the derivatized monosaccharide and/or polysaccharide components to differentially migrate within the electrophoretic medium. Then detecting the derivatized monosaccharide and/or polysaccharide components is accomplished by laser exciting the derivatized monosaccharide and/or polysaccharide components and monitoring fluorescent emission of the derivatized monosaccharide and/or polysaccharide components. The processes provide extremely low detection limits of the labelled components.

Abstract: A multiple capillary biochemical analyzer for sequencing DNA and performing other analyses, in which a set of capillaries extends from wells in a microtiter plate into a cuvette. In the cuvette the capillaries are held on fixed closely spaced centers by passing through a sandwich construction having a pair of metal shims which squeeze between them a rubber gasket, forming a leak proof seal for an interior chamber in which the capillary ends are positioned. Sheath fluid enters the chamber and entrains filament sample streams from the capillaries. The filament sample streams, and sheath fluid, flow through aligned holes in a barrier member spaced close to the capillary ends, into a collection chamber having a lower glass window. The filament streams are illuminated above the barrier member by a laser, causing them to fluoresce. The fluorescence is viewed end-on by a CCD camera chip located below the glass window.

Abstract: The present invention relates generally to any application involving the monitoring of signal arising from ions produced by electrospray or other high pressure (>100 torr) ion sources. The present invention relates specifically to an apparatus and method for the detection of ions emitted from a capillary electrophoresis (CE) system, liquid chromatography, or other small-scale separation methods. And further, the invention provides a very simple diagnostic as to the quality of the separation and the operation of an electrospray source.

Abstract: A noise-suppressing capillary separation system for detecting the real-time presence or concentration of an analyte in a sample is provided. The system contains a capillary separation means through which the analyte is moved, a coherent light source that generates a beam which is split into a reference beam and a sample beam that irradiate the capillary, and a detector for detecting the reference beam and the sample beam light that transmits through the capillary. The laser beam is of a wavelength effective to be absorbed by a chromophore in the capillary. The system includes a noise suppressing system to improve performance and accuracy without signal averaging or multiple scans.

Abstract: An apparatus and method of aligning a capillary with respect to a radiation source. More particularly, the capillary is aligned with a laser beam for laser induced fluorescence detection. The scatter or transmitted light pattern of the laser beam with respect to the capillary tube is utilized to determine optimum alignment. photosensors may be implemented to detect the light pattern which represents optimum alignment. For dynamic alignment during electrophoresis, the photosensors provide feedback to a controller which controls a positioning mechanism for alignment of the capillary.

Abstract: An electrophoresis apparatus includes a plurality of first capillaries containing a separation medium, such as a gel, and an optical cell in which one end of each of the first capillaries is disposed. Samples labeled with fluorophores are introduced into the first capillaries, and an electric field is applied to the first capillaries to cause the samples to migrate through the first capillaries into the optical cell. A light source excites the fluorophores with light when the samples are in the optical cell, causing the fluorophores to emit fluorescence, and a photodetecting system detects the fluorescence emitted by the fluorophores. The ends of the first capillaries in the optical cell may be arranged in a straight line. The light source may include a He-Ne laser emitting light having a wavelength of 594 nm, and the fluorophores may include either sulforhodamine or a derivative of sulforhodamine.

Abstract: A first reactant is immobilized i.e. in a porous matrix (50), adjacent a sample electrode (46) within a reaction chamber. Energizing of the electrode (46) electrophoretically attracts a mobile second reactant and/or electrolytically induces appropriate reaction conditions to enhance reaction of the first and second reactants. Polarity reversals between the sample electrode (46) and remote electrodes (38), (42), (44) cause unreacted second reactant and/or by-products to migrate away from the immobilized first reactant. The techniques are useful for sequential chemical reactions such as sequencing or construction of proteins, polysaccharides and nucleic acids where cyclical additions and removals of reactants are required. The techniques are amenable to automated micro and nano scale construction and operation and allow direct electrophoretic (38) interfacing with chromatographic, HPCE and mass spectrophotometric equipment.

Abstract: The invention relates to a device and a method for mass-spectrometric analysis of substance samples which have been separated by capillary electrophoresis, particularly proteins, proteoglycanes or other protein conjugates, with ionization of the substance samples by a particularly substance-saving form of electrospraying. A method of "microspraying" is used which permits a flow of spray liquid of only several tens of nanoliters per minute in stable spray operation. The invention consists in a loose coupling of the spray capillary to the electrophoresis capillary in a liquid environment.

Abstract: In many separation techniques, such as field flow fractionation, liquid chromatography and electro-phoresis, chemical species form bands that migrate at different velocities. If the data-digitization rate and excitation intensity are both set to be optimal for the fastest migrating band, to compensate for different band velocities, both the data-digitization rate and the excitation intensity are decreased as a function of time by a factor equal to the migration time of the fastest migrating band to the separation time.

Abstract: A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis.

Abstract: A method of quantitating proteins in complex samples using capillary electrophoresis can be used both to determine the concentration of a protein in a sample and to determine total protein concentration in the sample.

Abstract: A means and method for capillary zone electrphoresis with laser-induced indirect fluorescence detection. A detector is positioned on the capillary tube of a capillary zone electrophoresis system. The detector includes a laser which generates a laser beam which is imposed upon a small portion of the capillary tube. Fluorescence of the elutant electromigrating through the capillary tube is indirectly detected and recorded.