Source: http://www.google.com/patents/US20090146380?ie=ISO-8859-1&dq=4484186
Timestamp: 2014-03-15 00:05:34
Document Index: 562853467

Matched Legal Cases: ['Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60', 'Application No. 60']

Patent US20090146380 - Methods and apparatuses for generating a seal between a conduit and a ... - Google PatentsSearch Images Maps Play YouTube News Gmail Drive More »Sign inAdvanced Patent SearchPatentsMethods and Apparatuses for Generating a Seal Between a Conduit and a Reservoir Well. According to one embodiment, an apparatus is provided for generating a seal between a conduit and a reservoir well. The apparatus can include a mount including a first and second end. The mount can also include a first...http://www.google.com/patents/US20090146380?utm_source=gb-gplus-sharePatent US20090146380 - Methods and apparatuses for generating a seal between a conduit and a reservoir wellAdvanced Patent SearchPublication numberUS20090146380 A1Publication typeApplicationApplication numberUS 11/719,522PCT numberPCT/US2006/031249Publication dateJun 11, 2009Filing dateAug 10, 2006Priority dateAug 11, 2005Also published asWO2007021864A2, WO2007021864A3Publication number11719522, 719522, PCT/2006/31249, PCT/US/2006/031249, PCT/US/2006/31249, PCT/US/6/031249, PCT/US/6/31249, PCT/US2006/031249, PCT/US2006/31249, PCT/US2006031249, PCT/US200631249, PCT/US6/031249, PCT/US6/31249, PCT/US6031249, PCT/US631249, US 2009/0146380 A1, US 2009/146380 A1, US 20090146380 A1, US 20090146380A1, US 2009146380 A1, US 2009146380A1, US-A1-20090146380, US-A1-2009146380, US2009/0146380A1, US2009/146380A1, US20090146380 A1, US20090146380A1, US2009146380 A1, US2009146380A1InventorsGregory A. Votaw, Kelly Junge, Michael G. Pollack, Hugh C. CrenshawOriginal AssigneeEksigent Technologies, LlcExport CitationBiBTeX, EndNote, RefManReferenced by (1), Classifications (32), Legal Events (3) External Links: USPTO, USPTO Assignment, EspacenetMethods and apparatuses for generating a seal between a conduit and a reservoir wellUS 20090146380 A1Abstract Methods and Apparatuses for Generating a Seal Between a Conduit and a Reservoir Well. According to one embodiment, an apparatus is provided for generating a seal between a conduit and a reservoir well. The apparatus can include a mount including a first and second end. The mount can also include a first aperture extending between the first and second ends. The apparatus can also include a tube including a first end engaging the first end of the mount, and operable to hold a conduit having an end such that the conduit extends through the first aperture of the mount and the end of the conduit communicates with a reservoir well. Further, the apparatus can include a nut operable to engage the mount and tube and seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first aperture of the mount.
1. An apparatus for generating a seal between a conduit and a reservoir well, comprising:
(a) a mount including a first and second end, and including a first aperture extending between the first and second ends; (b) a tube including a first end engaging the first end of the mount, and operable to hold a conduit having an end such that the conduit extends through the first aperture of the mount and the end of the conduit communicates with a reservoir well; and (c) a nut operable to engage the mount and tube and seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first aperture of the mount. 2. The apparatus according to claim 1 wherein the first end of the mount comprises a threadable interior for engaging the nut.
3. The apparatus according to claim 2 wherein the nut includes a threadable exterior for engaging the threadable interior of the first end of the mount.
4. The apparatus according to claim 1 wherein the conduit comprises a fused silica capillary.
5. The apparatus according to claim 1 wherein the reservoir well comprises a covering, and comprising a needle attached to the mount and operable to penetrate the covering of the reservoir well and hold the end of the conduit in the reservoir well.
6. The apparatus according to claim 5 wherein the tube is a first tube, and comprising a rigid tube including a first end and second end, the first end of the rigid tube engaging the second end of the mount, the second end of the rigid tube extending into the needle, and operable to hold the conduit.
7. The apparatus according to claim 6 comprising a spring mechanism operable to provide resistance to movement of the mount with respect to the rigid tube and the first tube.
8. The apparatus according to claim 5 comprising an air pressure manifold communicating with the needle for communicating air from an air supply into the reservoir well such that fluid in the reservoir well is forced into the end of the conduit.
9. The apparatus according to claim 8 wherein the rigid tube extends through the air pressure manifold.
10. The apparatus according to claim 9 comprising an air-lock nut operable to engage the rigid tube and the air pressure manifold for forming a seal therebetween.
11. The apparatus of claim 1 comprising an o-ring operable to engage the mount and the needle for forming a seal therebetween.
12. The apparatus according to claim 1 wherein the reservoir well comprises an opening, and comprising:
(a) a needle attached to the mount and operable to hold the end of the conduit in the reservoir well; and (b) a gasket including an aperture operable to hold the needle in the reservoir well and sealingly engage the opening of the reservoir well. 13. The apparatus according to claim 1 comprising:
(a) a needle attached to the mount and operable to hold the end of the conduit in the reservoir well; (b) a gasket including an aperture operable to sealingly engage an opening of the reservoir well and position the needle in the reservoir well; and (c) a spring including a first and second end, the first end of the spring attached to the needle, and the second end of the spring attached to the gasket such that the gasket is evenly applied to the opening of the reservoir well when the needle is positioned in the reservoir well. 14. A method for generating a seal between a conduit and a reservoir well, the method comprising:
(a) providing a mount including a first and second end, and including a first aperture extending between the first and second ends; (b) providing a tube including a first end engaging the first end of the mount, and operable to hold a conduit having an end such that the conduit extends through the first aperture of the mount and the end of the conduit communicates with a reservoir well; (c) providing a nut operable to engage the mount and tube and seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first aperture of the mount; and (d) inserting the conduit into the tube such that the end of the tube is positioned in the reservoir well. 15. The method according to claim 14 wherein the first end of the mount comprises a threadable interior for engaging the nut.
16. The method according to claim 14 wherein the nut includes a threadable exterior for engaging the threadable interior of the first end of the mount.
17. The method according to claim 14 wherein the conduit comprises a fused silica capillary.
18. The method according to claim 14 wherein the reservoir well comprises a covering, and comprising:
(a) providing a needle attached to the mount and operable to hold the end of the conduit; and (b) inserting the needle into the covering of the reservoir well such that the end of the conduit is in the reservoir well. 19. The method according to claim 18 comprising providing a rigid tube including a first end and second end, the first end of the rigid tube engaging the second end of the mount, the second end of the rigid tube extending into the needle, and operable to hold the conduit.
20. The method according to claim 19 comprising providing an air pressure manifold communicating with the needle for communicating air from an air supply into the reservoir well such that fluid in the reservoir well is forced into the end of the conduit.
21. The method according to claim 20 wherein the rigid tube extends through the air pressure manifold.
22. The method according to claim 21 comprising providing an air-lock nut operable to engage the rigid tube and the air pressure manifold for forming a seal therebetween.
23. The method according to claim 14 wherein the reservoir well comprises and opening, and comprising:
(a) providing a needle attached to the mount and operable to hold the end of the conduit in the reservoir well; and (b) providing a gasket including an aperture operable to hold the needle in the reservoir well and sealingly engage the opening of the reservoir well. 24-39. (canceled)
40. An apparatus for generating a seal between a conduit and a reservoir well, comprising:
(a) a reservoir well comprising a covering; (b) a mount including a first and second end, and including an aperture extending between the first and second ends; (c) a conduit having an end such that the conduit extends through the first aperture of the mount, the end of the conduit communicates with the reservoir well; (d) a nut and tube operable to seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first aperture of the mount, and operable to permit the conduit to move with respect to the first aperture; and (e) a needle operable to penetrate the covering of the reservoir well, and hold an end of the tube in the reservoir well. 41-53. (canceled)
CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Patent Application Ser. No. 60/707,286, filed Aug. 11, 2005, the disclosure of which is incorporated herein by reference in its entirety. The disclosures of the following U.S. Provisional Applications, commonly owned and simultaneously filed Aug. 11, 2005, are all incorporated by reference in their entirety: U.S. Provisional Application entitled MICROFLUIDIC APPARATUS AND METHOD FOR SAMPLE PREPARATION AND ANALYSIS, U.S. Provisional Application No. 60/707,373 (Attorney Docket No. 447/99/2/1); U.S. Provisional Application entitled APPARATUS AND METHOD FOR HANDLING FLUIDS AT NANO-SCALE RATES, U.S. Provisional Application No. 60/707,421 (Attorney Docket No. 447/99/2/2); U.S. Provisional Application entitled MICROFLUIDIC BASED APPARATUS AND METHOD FOR THERMAL REGULATION AND NOISE REDUCTION, U.S. Provisional Application No. 60/707,330 (Attorney Docket No. 447/99/2/3); U.S. Provisional Application entitled MICROFLUIDIC METHODS AND APPARATUSES FOR FLUID MIXING AND VALVING, U.S. Provisional Application No. 60/707,329 (Attorney Docket No. 447/99/2/4); U.S. Provisional Application entitled MICROFLUIDIC SYSTEMS, DEVICES AND METHODS FOR REDUCING DIFFUSION AND COMPLIANCE EFFECTS AT A FLUID MIXING REGION, U.S. Provisional Application No. 60/707,220 (Attorney Docket No. 447/99/3/1); U.S. Provisional Application entitled MICROFLUIDIC SYSTEMS, DEVICES AND METHODS FOR REDUCING NOISE GENERATED BY MECHANICAL INSTABILITIES, U.S. Provisional Application No. 60/707,245 (Attorney Docket No. 447/99/3/2); U.S. Provisional Application entitled MICROFLUIDIC SYSTEMS, DEVICES AND METHODS FOR REDUCING BACKGROUND AUTOFLUORESCENCE AND THE EFFECTS THEREOF U.S. Provisional Application No. 60/707,386 (Attorney Docket No. 447/99/3/3); U.S. Provisional Application entitled MICROFLUIDIC CHIP APPARATUSES, SYSTEMS, AND METHODS HAVING FLUIDIC AND FIBER OPTIC INTERCONNECTIONS, U.S. Provisional Application No. 60/707,246 (Attorney Docket No. 447/99/4/2); U.S. Provisional Application entitled METHODS FOR CHARACTERIZING BIOLOGICAL MOLECULE MODULATORS, U.S. Provisional Application No. 60/707,328 (Attorney Docket No. 447/99/5/1); U.S. Provisional Application entitled METHODS FOR MEASURING BIOCHEMICAL REACTIONS, U.S. Provisional Application No. 60/707,370 (Attorney Docket No. 447/99/5/2); U.S. Provisional Application entitled METHODS AND APPARATUSES FOR REDUCING EFFECTS OF MOLECULE ADSORPTION WITHIN MICROFLUIDIC CHANNELS, U.S. Provisional Application No. 60/707,366 (Attorney Docket No. 447/99/8); U.S. Provisional Application entitled PLASTIC SURFACES AND APPARATUSES FOR REDUCED ADSORPTION OF SOLUTES AND METHODS OF PREPARING THE SAME, U.S. Provisional Application No. 60/707,288 (Attorney Docket No. 447/9919); U.S. Provisional Application entitled BIOCHEMICAL ASSAY METHODS, U.S. Provisional Application No. 60/707,374 (Attorney Docket No. 447/99/10); U.S. Provisional Application entitled FLOW REACTOR METHOD AND APPARATUS, U.S. Provisional Application No. 60/707,233 (Attorney Docket No. 447/99/11); and U.S. Provisional Application entitled MICROFLUIDIC SYSTEM AND METHODS, U.S. Provisional Application No. 60/707,384 (Attorney Docket No. 447/99/12).
TECHNICAL FIELD The present disclosure generally relates to microfluidic processing of reagents and analysis of reaction products. More specifically, the present disclosure relates to drawing regents from well plates.
One consideration when employing a microfluidic system to acquire data is minimizing carry-over in experiments that perform sequential analysis of liquids. The sequential analysis of liquids is central to the application of most analytical systems. For example, a microfluidic system that measures the potency of chemical inhibitors of an enzyme typically adds a sequence of different inhibitory compounds. Further, for example, microfluidic system diagnostic tests on blood must sequentially add different blood samples. Injection loops and automatic pipetting robots have been developed to permit sequential addition of liquids into an analytical system. An automatic pipetting robot can be used to add predefined volumes of fluid into a reaction vessel, sometimes including many parallel reaction vessels, such as microtiter plates. The pipetting portion of the robot can pick up one fluid and then another, adding each to its respective reaction vessel.
When liquids are sequentially analyzed, each liquid should be thoroughly removed from the system before subsequent liquids are added. The residual amount of a preceding liquid in the subsequent analysis is known as �carry-over�. The degree to which carry-over can be tolerated in the analytical system depends on the application. For chemical reactions, such as polymerase chain reaction (PCR), carry-over is not acceptable because this reaction is used to amplify the number of copies of DNA, and contaminating DNA will be faithfully amplified. For determining the potency of inhibitors of an enzymatic reaction, the carry-over can limit the dynamic range of the analytical system. Thus, if the carry-over is 1%, the dynamic range of the system is 100-fold (i.e., it can only measure inhibitors with potencies that range from an IC50 of X to an IC50 of 100X). If the system handles an inhibitor with an IC50 of X (i.e., it is a potent inhibitor because it inhibits at low concentration), then even a non-inhibiting compound next in the sequence will appear to have an IC50 of 100� (i.e., carryover of a potent inhibitor will make the next compound appear like a weaker inhibitor, even if the next compound is a non-inhibitor).
SUMMARY According to one embodiment, an apparatus for generating a seal between a conduit and a reservoir well is provided. The apparatus can include a mount including a first and second end. The mount can also include a first aperture extending between the first and second ends. The apparatus can also include a tube including a first end engaging the first end of the mount, and operable to hold a conduit having an end such that the conduit extends through the first aperture of the mount and the end of the conduit communicates with a reservoir well. Further, the apparatus can include a nut operable to engage the mount and tube and seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first aperture of the mount.
According to a second embodiment, a method for generating a seal between a conduit and a reservoir well is disclosed. The method can include a step for providing a mount including a first and second end. The mount can also include a first aperture extending between the first and second ends. The method can also include a step for providing a tube including a first end engaging the first end of the mount, and operable to hold a conduit having an end such that the conduit extends through the first aperture of the mount and the end of the conduit communicates with a reservoir well. Further, the method can include a step for providing a nut operable to engage the mount and tube and seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first aperture of the mount. The method can also include a step for inserting the conduit into the tube such that the end of the tube is positioned in the reservoir well.
According to a third embodiment, a pump assembly is disclosed. The pump assembly can include a thermal mass material defining an interior. Further, the pump assembly can include a pump positioned in the interior and adapted for fluid communicate fluid to a position outside of the interior.
According to a fourth embodiment, an apparatus for generating a seal between a conduit and a reservoir well is disclosed. The apparatus can include a mount including a first and second end, and including an aperture extending between the first and second ends. Further, the apparatus can include a conduit having an end such that the conduit extends through the first aperture of the mount. The end of the conduit can communicate with a reservoir well. The apparatus can also include a nut and tube operable to seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first aperture of the mount, and operable to permit the conduit to move with respect to the first aperture.
According to a fifth embodiment, a method for generating a seal between a conduit and reservoir well is disclosed. The method can include a step for providing a mount including a first and second end, and including an aperture extending between the first and second ends. Further, the method can include a step for providing a conduit having an end such that the conduit extends through the first aperture of the mount. The end of the conduit can communicate with a reservoir well. The method can also include a step for providing a nut and tube operable to seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first-aperture of the mount, and operable to permit the conduit to move with respect to the first aperture. In addition, the method can include a step for inserting the conduit into the mount such that the end of the conduit is positioned in the reservoir well.
According to a sixth embodiment, an apparatus for generating a seal between a needle and a reservoir well is disclosed. The apparatus can include a reservoir well comprising a covering. In addition, the apparatus can include a needle operable to penetrate the covering of the reservoir well, and hold an end of a tube in the reservoir well.
According to a seventh embodiment, a method for generating a seal between a needle and a reservoir well is disclosed. The method can include a step for providing a reservoir well comprising a covering. Further, the method can include a step for providing a needle operable to penetrate the covering of the reservoir well, and hold an end of a tube in the reservoir well. The method can also include a step for inserting the tube into the piercing needle such that the end of the tube is positioned in the reservoir well.
According to an eighth embodiment, an apparatus for generating a seal between a conduit and a reservoir well. The apparatus can include a reservoir well comprising a covering. In addition, the apparatus can include a mount including a first and second end. The mount can also include an aperture extending between the first and second ends. The apparatus can also include a conduit having an end such that the conduit extends through the first aperture of the mount. The end of the conduit can communicate with the reservoir well. Further, the apparatus can include a nut and tube operable to seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first aperture of the mount, and operable to permit the conduit to move with respect to the first aperture. The apparatus can also include a needle operable to penetrate the covering of the reservoir well, and hold an end of the tube in the reservoir well.
According to a ninth embodiment, a method for generating a seal between a conduit and a reservoir well is disclosed. The method can include a step for providing a reservoir well comprising a covering. In addition, the method can include a step for providing a mount including a first and second end, and including an aperture extending between the first and second ends. The method can also include a step for providing a conduit having an end such that the conduit extends through the first aperture of the mount, the end of the conduit communicates with the reservoir well. Further, the method can include a step for providing a nut and tube operable to seal the conduit to the first aperture of the mount such that air cannot communicate from the reservoir well through the first aperture of the mount, and operable to permit the conduit to move with respect to the first aperture. The method can also include a step for providing a needle operable to penetrate the covering of the reservoir well, and hold an end of the tube in the reservoir well. In addition, the method can include a step for inserting the tube through the conduit and piercing needle such that the end of the tube is positioned in the reservoir well.
Therefore, it is an object to provide methods and apparatuses for generating a seal between a conduit and a reservoir well.
FIG. 21E is a top plan view of another exemplary microfluidic chip;
As used herein, the term �microfluidic chip,� �microfluidic system,� or �microfluidic device� generally refers to a chip, system, or device which can incorporate a plurality of interconnected channels or chambers, through which materials, and particularly fluid borne materials can be transported to effect one or more preparative or analytical manipulations on those materials. A microfluidic chip is typically a device comprising structural or functional features dimensioned on the order of mm-scale or less, and which is capable of manipulating a fluid at a flow rate on the order of 111/min or less. Typically, such channels or chambers include at least one cross-sectional dimension that is in a range of from about 1 μm to about 500 μm. The use of dimensions on this order allows the incorporation of a greater number of channels or chambers in a smaller area, and utilizes smaller volumes of reagents, samples, and other fluids for performing the preparative or analytical manipulation of the sample that is desired.
Suitable examples of such a microfluidic chip MFC are disclosed in co-pending, commonly owned U.S. Provisional Applications entitled MICROFLUIDIC SYSTEMS, DEVICES AND METHODS FOR REDUCING DIFFUSION AND COMPLIANCE EFFECTS AT A FLUID MIXING REGION, U.S. Provisional Application No. 60/707,220 (Attorney Docket No. 447/99/3/1); MICROFLUIDIC SYSTEMS, DEVICES AND METHODS FOR REDUCING NOISE GENERATED BY MECHANICAL INSTABILITIES, U.S. Provisional Application No. 60/707,245 (Attorney Docket No. 447/99/3/2); MICROFLUIDIC SYSTEMS, DEVICES AND METHODS FOR REDUCING BACKGROUND AUTOFLUORESCENCE AND THE EFFECTS THEREOF, U.S. Provisional Application No. 60/707,386 (Attorney Docket No. 447/99/3/3); and MICROFLUIDIC CHIP APPARATUSES, SYSTEMS, AND METHODS HAVING FLUIDIC AND FIBER OPTIC INTERCONNECTIONS, U.S. Provisional Application No. 60/707,246 (Attorney Docket No. 447/99/4/2), the contents of which are incorporated herein in their entireties. As discussed therein, to provide internal channels, microfluidic chip MFC can comprise two body portions such as plates or layers, with one body portion serving as a substrate or base on which features such as channels are formed and the other body portion serving as a cover. The two body portions can be bonded together by any means appropriate for the materials chosen for the body portions. Non-limiting examples of bonding techniques include thermal bonding, anodic bonding, glass frit bonding, adhesive bonding, and the like. Non-limiting examples of materials used for the body portions include various structurally stable polymers such as polystyrene, metal oxides such as sapphire (Al2O3), silicon, and oxides, nitrides or oxynitrides of silicon (e.g., SixNy, glasses such as SiO2, or the like). In advantageous embodiments, the materials are chemically inert and biocompatible relative to the reagents to be processed, or include surfaces, films, coatings or are otherwise treated so as to be rendered inert and/or biocompatible. The body portions can be constructed from the same or different materials. To enable optics-based data encoding of analytes processed by microfluidic chip MFC, one or both body portions can be optically transmissive or include windows at desired locations. The channels can be formed by any suitable micro-fabricating techniques appropriate for the materials used, such as the various etching, masking, photolithography, ablation, and micro-drilling techniques available. The channels can be formed, for example, according to the methods disclosed in a co-pending, commonly owned U.S. Provisional Application entitled MICROFLUIDIC CHIP APPARATUSES, SYSTEMS, AND METHODS HAVING FLUIDIC AND FIBER OPTIC INTERCONNECTIONS, U.S. Provisional Application No. 60/707,246 (Attorney Docket No. 447/9914/2), the content of which is incorporated herein in its entirety. In some embodiments, the size of the channels can range from approximately 5 to 500 μm in cross-sectional area.
As further illustrated in FIG. 1, a detection location or point DP is defined in microfluidic chip MFC at an arbitrary point along the flow path of the reagent mixture, e.g., at a desired point along aging loop AL. More than one detection point DP can be defined so as to enable multi-point measurements and thus permit, for example, the measurement of a reaction product at multiple points along aging loop AL and hence analysis of time-dependent phenomena or automatic localization of the optimum measurement point (e.g., finding a point yielding a sufficient yet not saturating analytical signal). In some methods as further described hereinbelow, however, only a single detection point DP is needed. Detection point DP represents a site of microfluidic chip MFC at which any suitable measurement (e.g., concentration) of the reagent mixture can be taken by any suitable encoding and data acquisition technique. As one example, an optical signal can be propagated though microfluidic chip MFC at detection point DP, such as through its thickness (e.g., into or out from the sheet of FIG. 1) or across its plane (e.g., toward a side of the sheet of FIG. 1), to derive an analytical signal for subsequent off-chip processing. Hence, microfluid chip MFC at detection point DP can serve as a virtual, micro-scale flow cell as part of a sample analysis instrument.
Exemplary enzymological variables and measurements that can be analyzed and prepared include, but are not limited to:
(1) basic steady-state kinetic constants, such as Michaelis constants for substrates (Km), maximum velocity (Vmax), and the resultant specificity constant (Vmax/Km or kcat/Km);
(3) kinetic mechanism of a bi- or multi-substrate enzyme reaction;
(7) dose-response of inhibitor or activator on enzyme or receptor activity (IC50 and EC50 value);
(8) analysis of mechanism of inhibition of an enzyme catalyzed reaction and associated inhibition constants (slope inhibition constant (Kis) and intercept inhibition constant (Kii)); and
(9) equilibrium binding experiments to determine binding constants (Kd);
(10) determination of binding stoichiometry via a continuous variation method.
Generally, excitation source ES can be any suitable continuum or line source or combination of sources for providing a continuous or pulsed input of initial electromagnetic energy (hv)0 to detection point DP (FIG. 1) of microfluidic chip MFC. Non-limiting examples include lasers, such as visible light lasers including green HeNe lasers, red diode lasers, and frequency-doubled Nd:YAG lasers or diode pumped solid state (DPSS) lasers (532 nm); hollow cathode lamps; deuterium, helium, xenon, mercury and argon arc lamps; xenon flash lamps; quartz halogen filament lamps; and tungsten filament lamps. Broad wavelength emitting light sources can include a wavelength selector WS1 as appropriate for the analytical technique being implemented, which can comprise one or more filters or monochromators that isolate a restricted region of the electromagnetic spectrum. Upon irradiation of the sample at detection point DP, a responsive analytical signal having an attenuated or modulated energy (hv)1 is emitted from microfluidic chip MFC and received by radiation detector Rb. Any suitable light-guiding technology can be used to direct the electromagnetic energy from excitation source ES, through microfluidic chip MFC, and to the remaining components of the measurement instrumentation. In some embodiments, optical fibers are employed. The interfacing of optical fibers with microfluidic chip MFC according to advantageous embodiments is disclosed in a co-pending, commonly owned U.S. Provisional Application entitled MICROFLUIDIC CHIP APPARATUSES, SYSTEMS, AND METHODS HAVING FLUIDIC AND FIBER OPTIC INTERCONNECTIONS, U.S. Provisional Application No. 60/707,246 (Attorney Docket No. 447/99/4/2), the content of which is incorporated herein in its entirety. In some embodiments, a miniaturized dip probe can be employed at detection point DP, in which both the optical sending and returning fibers enter the same side of microfluidic chip MFC and a reflective element routes the optical signal down the sending fiber back through the microfluidic channel to the returning fiber. Similarly a single fiber can be used both to introduce the light and to collect the optical signal and return it to a detector. For example, the excitation light for a fluorophore can be introduced into the microfluidic chip by an optical fiber, and the fluorescent light emitted by the sample in the microfluidic chip can be collected by that same fiber and transmitted to a photodetector, with appropriate wavelength selectors permitting rejection of excitation light at the photodetector.
Wavelength selector WS2 is utilized as appropriate for the analytical technique being implemented, and can comprise one or more filters or monochromators that isolate a restricted region of the electromagnetic spectrum and provide a filtered signal (hv)z for subsequent processing. Radiation detector RD can be any appropriate photoelectric transducer that converts the radiant energy of filtered analytical signal (hv)2 into an electrical signal I suitable for use by signal processing and readout device SPR. Non-limiting examples include photocells, photomultiplier tubes (PMTs), avalanche photodiodes (APDs), photodiode arrays (PDAs), and charge-coupled devices (CCDs). In particular, for fluorescence measurements, a PMT or APD can be operated in a photon counting mode to increase sensitivity or yield improved signal-to-noise ratios. Advantageously, radiation detector RD is enclosed in an insulated and opaque box to guard against thermal fluctuations in the ambient environment and keep out light.
Signal processing and readout device SPR can perform a number of different functions as necessary to condition the electrical signal for display in a human-readable form, such as amplification (i.e., multiplication of the signal by a constant greater than unity), phase shifting, logarithmic amplification, rationing, attenuation (i.e., multiplication of the signal by a constant smaller than unity), integration, differentiation, addition, subtraction, exponential increase, conversion to AC, rectification to DC, comparison of the transduced signal with one from a standard source, and/or transformation of the electrical signal from a current to a voltage (or the converse of this operation). In addition, signal processing and readout device SPR can perform any suitable readout function for displaying the transduced and processed signal, and thus can include a moving-coil meter, a strip-chart recorder, a digital display unit such as a digital voltmeter or CRT terminal, a printer, or a similarly related device. Finally, signal processing and readout device SPR can control one or more other components of sample processing apparatus SPA as necessary to automate the mixing, sampling/measurement, and/or temperature regulation processes of the methods disclosed herein. For instance, signal processing and readout device SPR can be placed in communication with excitation source ES, pumps PA-PC and thermal control unit TCU via suitable electrical lines to control and synchronize their respective operations, as well as receive feedback from the encoders typically provided with pumps PA-PC.
In advantageous embodiments, pump assembly PA provides temperature-control functionality. While both heating and cooling can be effected, the ability to cool pump assembly PA is particularly advantageous as it enables thermally labile reagents to be cooled in-situ to prevent their degradation, thereby eliminating the need for ex-situ or on-chip refrigeration. Proteins, for example, can denature at room temperatures in a matter of hours. Thus, cooling is particularly important when lengthy run times are contemplated. For example, if a 10-μl barrel is used, approximately 8 hours of run time is possible at a flow rate of 20 nl/min. In one embodiment, pump assembly PA can maintain a reagent temperature ranging from approximately −4� C. to 70� C. to within 0.05� C. of accuracy. Moreover, thermal control of pump assembly PA provides the flow stability and noise reduction needed when operating at flow rates in the nl/min range. A change in room temperature can cause thermal expansion of the components of pump assembly PA that interact with the liquids being conveyed, thereby causing a thermal pumping effect. For example, when pumping at a low flow rate such as a few nl/min, a 1-nl change in the volume of the system (i.e., 0.01 percent of total volume for a 10 μl syringe pump) over one minute will be noticeable. Similarly, a 1� C. change in the temperature of the stainless steel plunger of some microsyringes causes the plunger to change length by 2 μm, changing the volume inside the microsyringe by 0.3 nl. Because room temperature is a disturbance, thermal pumping appears as noise in the output of the pumps of pump assembly PA. Hence, controlling the temperature of pump assembly PA reduces this noise. Finally, with regard to the multi-pump configuration illustrated in FIGS. 7A-7C, the ability to regulate all pumps PA-PD at the same temperature reduces any disparity in any temperature gradients respectively existing between each pump PA-PD. Otherwise, the existence of different temperature gradients between pumps PA-PD can cause pumps PAPD to thermally pump out of phase with each other, which can also contribute to signal noise.
First annular member 202 has a bore 202A large enough to receive pump barrel 22. Hollow gasket 208 is sized to effect a fluid seal between pump barrel 22 and female fitting 210 when inserted into bore 202A of first annular member 202, Hollow gasket 208 is inserted far enough to abut the distal end of pump barrel 22, and has a bore 208A fluidly communicating with that of pump barrel 22 and aperture 210C of female fitting 210. In some embodiments, hollow gasket 208 is constructed from polytetrafluoroethylene (PTFE). Second annular member 204 is coaxially disposed about first annular member 202, and is removably secured thereto such as by providing mating threads on an outside surface 202B of first annular member 202 and an inside surface 204A of second annular member 204. Female fitting 210 is disposed within a cavity 206A of third annular member 206 and extends through a bore 206B of third annular member 206. The proximal end of female fitting 210, which can be defined by a flanged portion thereof, abuts the distal end of hollow gasket 208 and may abut the distal ends of first annular member 202 and/or second annular member 204. Female fitting 210 has a bore 210B beginning at a proximal aperture 210C disposed in axial alignment with bore 208A of hollow gasket 208. In the illustrated embodiment, at least a portion of bore 210B of female fitting 210 is tapered, and this tapered profile is complementary to a tapered profile presented by an outside surface 212A of male fitting 212 to effect a removable seal interface.
Connection of external pumps PAPD to microfluidic chip MFC and to external components, such as switching valves and plate handlers as discussed below, requires the use of tubes or other conduits. These should be of minimal internal volume for efficient use of reagents, and their walls should have minimal compliance to avoid their behaving like a pressure �capacitor� in which the walls expand (and thus the internal volume increases) as pressure increases to drive fluid flows. Materials such as fused silica can be readily obtained as microcapillaries with small internal diameters and rigid walls. Additionally, the capillaries should be shielded from thermal fluctuations because thermal expansion of the capillaries will cause them to behave like thermal pumps, and oscillations in temperature will result in noise in the flows through these capillaries. Such shielding can be either as an insulative wrap around the capillaries, or all components of the system, including the capillaries, can be housed in a single temperature-controlled enclosure.
Referring now to FIGS. 15A-15C, non-limiting examples of liquid handling systems are illustrated. These systems can be implemented with pump assembly PA in accordance with any of the embodiments of sample processing apparatus SPA disclosed herein. The automation provided by these systems offers many advantages. First, the automation can allow unattended refill of reagents in pumps PAPD, thus enabling the system to run unattended without operator intervention for days at a time. Second, the automation can allow automatic change of reagent in pumps PA-PD, and thus allow the system to test a series of reagents such as in screening pharmaceutical compounds, as well as the automatic reconfiguration of loaded reagents to automatically test the network of hypotheses for automated assay development and automatic hypothesis testing with intelligent systems. The automation also reduces the frequency that operators need to make and break fluidic interconnects. Thus, contamination and air bubbles in the system can be reduced, and the service life of the fluidic interconnects extended. These systems can incorporate an automated liquid handler that can be computer controlled via integrated computer software as part of any embodiment of the microfluidic systems disclosed herein. Managing the microfluidic system with a single software package enables real time decision-making and feedback control, thereby giving the system unprecedented flexibility and run time. This approach has not heretofore been practicable for displacement flows, because of the absence of displacement pumps that pump slowly enough for microfluidic systems as discussed hereinabove. An example of a suitable automated liquid handling system is the FAMOS� micro autosampler available from LC Packings, Sunnyvale, Calif. This system provides for automated sample injection of any volume ranging from 50 nl up to 25 μl from 96- and 384-well plates. The device can include a sample tray that is equipped with Peltier cooling to avoid degradation of thermally labile samples.
When switching valve SV switches to position 2, one of pumps PA, PB, PC and PD can be connected through injection loop INL to microfluidic chip MFC. One of pumps PA, PB, PC and PD can advance fluid from injection loop INL through a corresponding capillary IA, IB, IC and ID into microfluidic chip MFC. Simultaneously, the carriage can move capillary 274 to a well of multi-well plate MWP having a rinsing fluid. Syringe pump SP can then repeatedly pull fluid into and then expel fluid from capillary 274 to rinse it clean.
Carry-over can occur as different fluids are added into a microfluidic chip, such as microfluidic chip MFC shown in FIGS. 15A-15C. Carry-over can become greater as the volumetric flow rate through the microfluidic chip decreases, and can become extremely problematic at the very low flow rates desired for microfluidic systems, such as 30 nl/min. This is because the volumes displaced through the system are small relative to the volumes contained in the system. For example, the internal volume (sometimes referred to as �dead space�) of the smallest commercially available switching valve is 28 nl�Model CN2 switching valve from Valco Instrument Company of Houston, Tex., U.S.A. Thus, any void volumes or sources of contamination, which would be insignificant for faster flows that displace larger volumes per unit time, are now significant and, frequently, debilitating.
20-140 seconds: Pump PD=0 nl/minute, Pump PC=15 nl/minute 140-260 seconds: Pump PD increases linearly to 15 nl/minute, Pump PC decreases linearly to 0 nl/minute 260-380 seconds: Pump PD=15 nl/minute, Pump PC=0 nl/minute Pump PB flowed at a constant 10 nl/minute throughout.
FIGS. 19A-19C illustrate different views of a fluid freeze valve, generally designated FFVS, applied to a fluid-carrying capillary IL. Referring specifically to FIG. 19A, a top perspective view of fluid freeze valve FFVS is illustrated. Fluid freeze valve FFVS can include a movable top plate MTP and a thermoelectric cooler TEC (such as the Peltier Temperature Controller available from Stable Micro Systems Ltd. of London, England). Movable top plate MTP can be rotatably movable with respect to thermoelectric cooler TEC such that capillary IL can be positioned between movable top plate MTP and thermoelectric cooler TEC. FIG. 19B illustrates a side cross-sectional view of movable top plate MTP, thermoelectric cooler TEC, and capillary IL wherein thermo-electric cooler TEC is not energized such that fluid F can flow through lumen L of capillary IL in the �on� state. Movable top plate MTP can be made of a material having low thermal mass, low thermal conductivity, and does not absorb water. Movable top plate MTP can form an airtight seal around thermo-electric cooler TEC, or the assembly can be placed in an air-tight, low humidity chamber, such that water from the atmosphere does not condense onto thermoelectric cooler TEC, thereby adding thermal mass. FIG. 19C illustrates a side cross-sectional view of movable top plate MTP, thermoelectric cooler TEC, and capillary IL wherein thermo-electric cooler TEC is energized for reducing the temperature of capillary IL such that fluid F reaches a solid or nearly solid state to stop fluid flow through lumen L of capillary IL in the �off� state. Thermo-electric cooler TEC can also apply heat to capillary IL such that fluid F in a frozen or nearly frozen state can rapidly thaw, thereby returning the fluid freeze valve FFVS to the �on� state.
FIGS. 20A, 20B, and 20C illustrates a top, front and side view, respectively, of another fluid freeze valve, generally designated FFVS, applied to a fluid-carrying capillary IL. Fluid freeze valve FFVS can include a thermo-electric cooler TEC for application to a capillary IL. Thermo-electric cooler TEC can be attached to a heat sink HS containing a circulating water heat exchanger for removing heat from thermoelectric cooler TEC. Heat sink HS can also include tubes T1 and T2 for delivering and returning fluid to a liquid chiller (not shown). Tubes T1 and T2 can be connected to heat sink HS via quick-connects QC1 and QC2, respectively. The assembly can be mounted into a mounting plate MP for mounting to external supports.
Referring to FIG. 20A, fluid freeze valve FFVS can include an insulated housing surrounding thermoelectric cooler TEC comprising a removable top plate RTP lined on its internal surface with a conformal thermal insulation CTI that both pushes capillary IL against the surface of thermoelectric cooler TEC and thermally isolates capillary IL and thermoelectric cooler TEC from oscillations in ambient temperature. Similarly, the sides of thermoelectric cooler TEC can be surrounded by thermal insulation TI to further thermally isolate capillary IL and thermo-electric cooler TEC. Insulation can be important when a freeze valve is used to control low flow rates, such as of the nanoliter/minute scale. This can be important because water increases with volume when it freezes. For example, a thermoelectric cooler (such as thermoelectric cooler TEC shown in FIG. 20) of about 2 centimeters across can freeze about 2 centimeters of fluid in a capillary. If the capillary has an internal diameter of 50 micrometers, two centimeters of this capillary confines about 20 nanoliters. A length of 1 millimeter encloses about 2.0 nanoliters. Water increases volume about 9% when it freezes. If the edges of the frozen volume of fluid move 1 millimeter due to oscillations of ambient temperature that can affect either the temperature of the capillary or the temperature of thermoelectric cooler TEC, then the fluid adjacent to the frozen plug of fluid will change volume by about 0.14 nanoliters. For example, for flows of about 15 nanoliters/minute, such a 1 mm thaw over 1 minute represents a variation of more than 1%. Note that a capillary IL having a larger internal diameter can have a larger volume per unit length, so in the case where the fluid thaws over a fixed length, then a capillary having a larger diameter may introduce more noise to the flow.
Fluid freeze valves (such as fluid freeze valves FFVS shown in FIGS. 19A-19C and 20A-20C) can be applied to the systems described herein for stopping flow in a capillary attached to a microfluidic chip. For example, a fluid freeze valve can be applied to a capillary connecting a microsyringe pump and a microfluidic chip, a capillary connecting a microsyringe pump and an outside reservoir, or a capillary connecting a microfluidic chip and an outside multi-well plate or reservoir. It is important that the connection between the capillary and the microfluidic chip have minimal dead volume and minimal void volume, or carry-over may be increased.
FIGS. 21A-21D illustrate top plan views of different stages in a sample process run by a microfluidic system, generally designated MS. Microfluidic system MS can include a microfluidic chip MFC having injection loop INL and a plurality of fluid freeze valves VS1, VS2, and VS3. Injection loop INL can comprise a microchannel etched in microfluidic chip MFC having dimensions of about 150 micrometers wide, 150 micrometers deep, and 2 centimeters long for yielding a volume of 450 nanoliter. Alternatively, microchannel can have other suitable dimensions for achieving a desired volume. Microfluidic chip MFC can include a first and second input channel CH1 and CH2 for fluidly connecting or communicating at a merge point ML for combining fluids advanced therein from microsyringe pumps MP1 and MP2, respectively. Injection loop INL can be fluidly connected at one end to capillary CP1 and at an opposing end to capillary CP2. Capillaries CP1 and CP2 can be made of fused silica with 150 micrometers outside diameter and 75 micrometers inside diameter, respectively, available from Polymicro Technologies LLC. of Phoenix, Ariz. Capillaries can be connected to chips in accordance with embodiments disclosed in co-pending, commonly owned U.S. Provisional Application entitled MICROFLUIDIC CHIP APPARATUSES, SYSTEMS, AND METHODS HAVING FLUIDIC AND FIBER OPTIC INTERCONNECTIONS, U.S. Provisional Application No. 60/707,246 (Attorney Docket No. 447/9914/2), the content of which is incorporated herein in its entirety.
FIG. 21E is a top plan view of another exemplary microfluidic chip, generally designated MFC, having an injection loop INL; interconnect channels IC1, IC2, and IC3 for connecting to capillaries that connect to microsyringe pumps MP1, MP2, and MP3, respectively (FIG. 21); an interconnect channel ICCP3 that can connect to output capillary CP3 (FIG. 21); an interconnect channels ICCP1 and ICCP2 that can connect to capillaries CP1 and CP2, respectively (FIG. 21); an aging loop AL; and fiducial marks (F1, F2, and F3) for automated alignment.
FIGS. 25, 26A, 26B, and 26C illustrate cross-sectional views of different configurations for forming seals S1 and S2 shown in FIG. 23. Referring to FIG. 25, a cross-sectional view of a configuration for forming seal S1 is illustrated. Seal S1 can be formed when rubber septum RS is positioned to cover well W of multi-well plate MWP. According to one embodiment, foot SLF is a circular foot that presses uniformly onto septum RS such that seal S2 between septum RS and multi-well plate MWP can withstand the pressure. Seal S2 between septum RS and needle PN can be formed by the action of needle PN piercing septum RS. Thus, seal S2 can be formed by septum RS that is pushed by foot SLF.
In some instances, fluid in an injection loop (such as injection loop INL shown in FIGS. 21A-21D) should be maintained at a temperature different than that of an aging loop (such as aging loop AL shown in FIGS. 21A-21D). For example, a biochemical assay should be run at 370 Celsius in the aging loop while the fluid in the injection loop should be stored at 40 Celsius until the fluid enters the aging loop. FIGS. 28A and 28B illustrate schematic views of different microfluidic systems, generally designated MS, for maintaining fluids in an injection loop INL and aging loop AL at different temperatures. Microfluidic system MS can include a microfluidic chip MFC, a waste unit WU, a vacuum unit VU, a multi-well plate MWP, microsyringe pumps MP1, MP2, and MP3, and an injection loop INL. Waste unit WU can be connected to aging loop AL via a capillary CP1. Vacuum unit VU and multi-well plate MWP can be connected to injection loop INL via capillaries CP2 and CP3, respectively. Microfluidic system MS can also include fluid freeze valves VS1, VS2, and VS3 connected to capillaries CP1, CP2, and CP3, respectively.
Referring specifically to FIG. 28A, injection loop INL can comprise channels CH1 and CH2 in microfluidic chip MFC and a capillary CP4. Channels CH1 and CH2 and capillary CP4 can form injection loop INL and fluidly connect microsyringe MP3 and multi-well plate MWP at one end of injection loop INL to vacuum unit VU, aging loop AL, waste unit WU, and microsyringe pumps MP1 and MP2 at an opposing end of injection loop INL. Microfluidic system MS can also include a temperature control device TCD (such as the Peltier thermoelectric device) connected to a portion of capillary CP4 for cooling the fluid in that portion of capillary CP4. Temperature control device TCD can maintain the fluid at a desired temperature such as a desired temperature lower than the fluid in aging loop AL. Capillaries can be connected to chips in accordance with embodiments disclosed in a co-pending, commonly owned U.S. Provisional Application entitled MICROFLUIDIC CHIP APPARATUSES, SYSTEMS, AND METHODS HAVING FLUIDIC AND FIBER OPTIC INTERCONNECTIONS, U.S. Provisional Application No. 60/707,246 (Attorney Docket No. 447199/4/2), the content of which is incorporated herein in its entirety.
FIG. 28B illustrates a schematic diagram of microfluidic chip MFC having a portion containing injection loop INL that extends into temperature control device TCD. In this embodiment, injection loop INL is contained entirely on-chip and is located to a side portion of microfluidic chip MFC attached to temperature control unit TCU.
Referenced byCiting PatentFiling datePublication dateApplicantTitleUS8037788 *Jul 24, 2008Oct 18, 2011Dionex CorporationTight-spot fitting and driver, and method of use thereof* Cited by examinerClassifications U.S. Classification277/314, 277/628, 285/89, 277/625International ClassificationB01L99/00, F16J15/00, F16L21/02Cooperative ClassificationB01F13/0059, B01L3/502738, B01L2300/044, B01L2300/1822, B01L3/565, B01L3/50273, B01F5/0653, B01L2400/0487, B01L3/502715, B01L2300/1883, B01L3/502707, F04B19/006, B01L2300/0816, B01F5/0471, B01L2400/0677, B01L2300/1894, B01F5/0646European ClassificationB01F5/06B3F12, B01L3/5027B, B01L3/5027A, B01L3/565, B01F5/04C14, B01F5/06B3F, B01F13/00M, F04B19/00MLegal EventsDateCodeEventDescriptionMar 12, 2010ASAssignmentOwner name: AB SCIEX LLC,MASSACHUSETTSFree format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:EKSIGENT TECHNOLOGIES, LLC;US-ASSIGNMENT DATABASE UPDATED:20100312;REEL/FRAME:24066/427Effective date: 20100212Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:EKSIGENT TECHNOLOGIES, LLC;US-ASSIGNMENT DATABASE UPDATED:20100413;REEL/FRAME:24066/427Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:EKSIGENT TECHNOLOGIES, LLC;REEL/FRAME:024066/0427Owner name: AB SCIEX LLC, MASSACHUSETTSMar 6, 2007ASAssignmentOwner name: EKSIGENT TECHNOLOGIES, LLC, CALIFORNIAFree format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:SMITHKLINE BEECHAM CORPORATION;REEL/FRAME:018962/0919Effective date: 20070305Owner name: EKSIGENT TECHNOLOGIES, LLC,CALIFORNIAFree format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:SMITHKLINE BEECHAM CORPORATION;US-ASSIGNMENT DATABASE UPDATED:20100413;REEL/FRAME:18962/919Dec 11, 2006ASAssignmentOwner name: SMITHKLINE BEECHAM CORPORATION, PENNSYLVANIAFree format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:VOTAW, GREGORY A;JUNGE, KELLY;POLLACK, MICHAEL G.;AND OTHERS;REEL/FRAME:018610/0762;SIGNING DATES FROM 20061201 TO 20061207RotateOriginal ImageGoogle Home - Sitemap - USPTO Bulk Downloads - Privacy Policy - Terms of Service - About Google Patents - Send FeedbackData provided by IFI CLAIMS Patent Services©2012 Google