US20070020147A1 - Miniaturized fluid delivery and analysis system - Google Patents
Miniaturized fluid delivery and analysis system Download PDFInfo
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- US20070020147A1 US20070020147A1 US11/505,762 US50576206A US2007020147A1 US 20070020147 A1 US20070020147 A1 US 20070020147A1 US 50576206 A US50576206 A US 50576206A US 2007020147 A1 US2007020147 A1 US 2007020147A1
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502738—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by integrated valves
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F04—POSITIVE - DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS FOR LIQUIDS OR ELASTIC FLUIDS
- F04B—POSITIVE-DISPLACEMENT MACHINES FOR LIQUIDS; PUMPS
- F04B43/00—Machines, pumps, or pumping installations having flexible working members
- F04B43/02—Machines, pumps, or pumping installations having flexible working members having plate-like flexible members, e.g. diaphragms
- F04B43/04—Pumps having electric drive
- F04B43/043—Micropumps
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
- B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0883—Serpentine channels
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0887—Laminated structure
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
- B01L2400/0481—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure squeezing of channels or chambers
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/06—Valves, specific forms thereof
- B01L2400/0605—Valves, specific forms thereof check valves
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/06—Valves, specific forms thereof
- B01L2400/0633—Valves, specific forms thereof with moving parts
- B01L2400/0638—Valves, specific forms thereof with moving parts membrane valves, flap valves
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
- Y10T436/2575—Volumetric liquid transfer
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Clinical Laboratory Science (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Dispersion Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Mechanical Engineering (AREA)
- General Engineering & Computer Science (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention provides a method for combining a fluid delivery system with an analysis system for performing immunological or other chemical of biological assays. The method comprises a miniature plastic fluidic cartridge containing a reaction chamber with a plurality of immobilized species, a capillary channel, and a pump structure along with an external linear actuator corresponding to the pump structure to provide force for the fluid delivery. The plastic fluidic cartridge can be configured in a variety of ways to affect the performance and complexity of the assay performed.
Description
- This application claims priority to U.S. patent application Ser. No. 10/437,046, filed May 14, 2003, which is hereby incorporated by reference herein in its entirety.
- This invention relates to a system comprising a fluid delivery and analysis cartridge and an external linear actuator. More particularly, the invention relates to a system for carrying out various processes, including screening, immunological diagnostics, DNA diagnostics, in a miniature fluid delivery and analysis cartridge.
- Recently, highly parallel processes have been developed for the analysis of biological substances such as, for example, proteins and DNA. Large numbers of different binding moieties can be immobilized on solid surfaces and interactions between such moieties and other compounds can be measured in a highly parallel fashion. While the sizes of the solid surfaces have been remarkably reduced over recent years and the density of immobilized species has also dramatically increased, typically such assays require a number of liquid handling steps that can be difficult to automate without liquid handling robots or similar apparatuses.
- A number of microfluidic platforms have recently been developed to solve such problems in liquid handling, reduce reagent consumptions, and to increase the speed of such processes. Examples of such platforms are described in U.S. Pat. Nos. 5,856,174 and 5,922,591. Such a device was later shown to perform nucleic acid extraction, amplification and hybridization on HIV viral samples as described by Anderson et al, “Microfluidic Biochemical Analysis System”, Proceeding of the 1997 International Conference on Solid-State Sensors and Actuators, Tranducers '97, 1997, pp. 477-480. Through the use of pneumatically controlled valves, hydrophobic vents, and differential pressure sources, fluid reagents were manipulated in a miniature fluidic cartridge to perform nucleic acid analysis.
- Another example of such a microfluidic platform is described in U.S. Pat. No. 6,063,589 where the use of centripetal force is used to pump liquid samples through a capillary network contained on compact-disc liquid fluidic cartridge. Passive burst valves are used to control fluid motion according to the disc spin speed. Such a platform has been used to perform biological assays as described by Kellog et al, “Centrifugal Microfluidics: Applications,” Micro Total Analysis System 2000, Proceedings of the uTas 2000 Symposium, 2000, pp. 239-242. The further use of passive surfaces in such miniature and microfluidic devices has been described in U.S. Pat. No. 6,296,020 for the control of fluid in micro-scale devices.
- An alternative to pressure driven liquid handling devices is through the use of electric fields to control liquid and molecule motion. Much work in miniaturized fluid delivery and analysis has been done using these electro-kinetic methods for pumping reagents through a liquid medium and using electrophoretic methods for separating and perform specific assays in such systems. Devices using such methods have been described in U.S. Pat. No. 4,908,112, U.S. Pat. No. 6,033,544, and U.S. Pat. No. 5,858,804.
- Other miniaturized liquid handling devices have also been decribed using electrostatic valve arrays (U.S. Pat. No. 6,240,944), Ferrofluid micropumps (U.S. Pat. No. 6,318,970), and a Fluid Flow regulator (U.S. Pat. No. 5,839,467).
- The use of such miniaturized liquid handling devices has the potential to increase assay throughput, reduce reagent consumption, simplify diagnostic instrumentation, and reduce assay costs.
- The system of the invention comprises a plastic fluidic device having at least one reaction chamber connected to pumping structures through capillary channels and external linear actuators. The device comprises two plastic substrates, a top substrate and a bottom substrate containing capillary channel(s), reaction chamber(s), and pump/valve chamber(s)—and a flexible intermediate interlayer between the top and bottom substrate which provides providing a sealing interface for the fluidic structures as well as valve and pump diaphragms. Passive check valve structures are formed in the three layer device by providing a means for a gas or liquid to flow from a channel in the lower substrate to a channel in the upper substrate by the bending of the interlayer diaphragm. Furthermore flow in the opposite direction is controlled by restricting the diaphragm bending motion with the lower substrate. Alternatively check valve structures can be constructed to allow flow from the top substrate to the bottom substrate by flipping the device structure. Pump structures are formed in the device by combining a pump chamber with two check valve structures operating in the same direction. A hole is also constructed in the lower substrate corresponding to the pump chamber. A linear actuator—external to the plastic fluidic device—can then be placed in the hole to bend the pump interlayer diaphragm and therefore provide pumping action to fluids within the device. Such pumping structures are inherently unidirectional.
- In one embodiment the above system can be used to perform immunoassays by pumping various reagents from an inlet reservoir, through a reaction chamber containing a plurality of immobilized antibodies or antigens, and finally to an outlet port. In another embodiment the system can be used to perform assays for DNA analysis such as hybridization to DNA probes immobilized in the reaction chamber. In still another embodiment the device can be used to synthesize a series of oligonucleotides within the reaction chamber. While the system of the invention is well suited to perform solid-phase reactions within the reaction chamber and provide the means of distributing various reagents to and from the reaction chamber, it is not intended to be limited to performing solid-phase reactions only.
- The system of the invention is also well suited for disposable diagnostic applications. The use of the system can reduce the consumables to only the plastic fluidic cartridge and eliminate any cross contamination issues of using fixed-tipped robotic pipettes common in high-throughput applications.
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FIG. 1A is a top view of a pump structure within the plastic fluidic device of the invention. -
FIG. 1B is a cross section view of the pump structure within the plastic fluidic device of the invention. -
FIG. 2 is a top view of a plastic fluidic device of the invention configured as a single-fluid delivery and analysis device. -
FIG. 3 is a top view of a plastic fluidic device of the invention configured as a 5-fluid delivery and analysis device. -
FIG. 4 is a top view of a plastic fluidic device of the invention configured as a re-circulating 3-fluid delivery and analysis device. - The system of the invention comprises a plastic fluidic cartridge and a linear actuator system external to the fluidic cartridge.
FIG. 1A shows a cross-sectional view of a pump structure formed within the fluidic cartridge of the invention. The plastic fluidic cartridge comprises three primary layers: anupper substrate 21, alower substrate 22, and a flexibleintermediate interlayer 23, as shown inFIG. 1B . The three layers can be assembled by various plastic assembly methods such as, for example, screw assembly, heat staking, ultrasonic bonding, clamping, or suitable reactive/adhesive bonding methods. The upper and lower substrates, depicted as 21 and 22 inFIG. 1B , both contain a variety of features that define channels of capillary dimensions as well as pump chambers, valve chambers, reaction chambers, reservoirs, and inlet/outlet ports within the cartridge.FIG. 1B shows a top view of the pump structure ofFIG. 1A . The pump is defined by apump chamber 14 and twopassive check valves 15 that provide a high resistance to flow in one direction only.Passive check valves 15 comprise alower substrate channel 13 and anupper substrate channel 11 separated byinterlayer 23 such that holes throughinterlayer 23, depicted asholes 12 inFIG. 1B , are contained withinupper substrate channel 11 but not withinlower substrate channel 13. Such check valve structures provide a low resistance to a gas/liquid flowing fromlower substrate channel 13 toupper substrate channel 11 and likewise provide a high resistance to a gas/liquid flowing fromupper substrate channel 11 tolower substrate channel 13.Pump chamber 14 comprises an upper substrate chamber and ahole 141 inlower substrate 22 tofree interlayer 23 to act as adiaphragm 25, as depicted inFIG. 1B . Alinear actuator 24 external to the fluidic cartridge can then be placed in the hole 131 to benddiaphragm 25 and therefore provide the necessary force to deform the diaphragm. -
FIG. 2 shows a top view of a plastic fluidic cartridge of the invention configured as a single-fluid delivery and analysis device. Fluid is first placed into thereservoir 31 manually or automated using a pipette or similar apparatus. Apump structure 32 similar to that ofFIG. 1B is contained within the device. By repeatedly actuating an external linear actuator, fluid inreservoir 31 is pumped through thepump structure 32, thecapillary channel 33 and into thereaction chamber 34.Reaction chamber 34 contains a plurality of immobilizedbio-molecules 35 for specific solid-phase reactions with said fluid. After a specified reaction time, the fluid is pumped throughreaction chamber 34 and out theexit port 36. -
Upper substrate 21 andlower substrate 22 of the plastic fluidic cartridge of the invention can be constructed using a variety of plastic materials such as, for example, polymethyl-methacrylate (PMMA), polystyrene (PS), polycarbonate (PC), Polypropylene (PP), polyvinylchloride (PVC). In the case of optical characterization of reaction results within a reaction chamber,upper substrate 21 is preferably constructed out of a transparent plastic material. Capillaries, reaction chambers, and pump chambers can be formed inupper substrate 21 andlower substrate 22 using methods such as injection molding, compression molding, hot embossing, or machining. Thicknesses ofupper substrate 21 andlower substrate 22 are suitably in, but not limited to, the range of 1 millimeter to 3 millimeter in thickness.Flexible interlayer 23 can be formed by a variety of polymer and rubber materials such as latex, silicone elastomers, polyvinylchloride (PVC), or fluoroelastomers. Methods for forming the features ininterlayer 23 include die cutting, rotary die cutting, laser etching, injection molding, and reaction injection molding. -
Linear actuator 24 of the present invention, as depicted inFIG. 1B , is preferred to be, but not limited to, an electromagnetic solenoid. Other suitable linear actuators include a motor/cam/piston configuration, a piezoelectric linear actuator, or motor/linear gear configuration. - The plastic fluidic cartridge, as shown in
FIG. 2 , can be utilized to perform immunological assays withinreaction chamber 34 by immobilizing a plurality of bio-molecules such asdifferent antibodies 35. In one exemplary embodiment, a sample containing an unknown concentration of a plurality of antigens or antibodies is first placed withinreservoir 31. The external linear actuator is then repeatedly actuated to pump the sample fromreservoir 31 toreaction chamber 34. The sample is then allowed to react with the immobilizedantibodies 35 for a set reaction time. At the end of the set reaction time, the sample is then excluded fromreaction chamber 34 throughexit port 36. A wash buffer is then placed inreservoir 31 and the external linear actuator is repeatedly actuated to pump the wash buffer throughreaction chamber 34 and out theexit port 36. Such wash steps can be repeated as necessary. A solution containing a specific secondary antibody conjugated with a detectable molecule such as a peroxidase enzyme, alkaline phosphatase enzyme, or fluorescent tag is placed intoreservoir 31. The secondary antibody solution is then pumped intoreaction chamber 34 by repeatedly actuating the linear actuator. After a predetermined reaction time, the solution is pumped out throughexit port 36.Reaction chamber 34 is then washed in a similar manner as previously describe. In the case of an enzyme conjugate, a substrate solution is placed intoreservoir 31 and pumped intoreaction chamber 34. The substrate will then react with any enzyme captured by the previous reactions with the immobilizedantibodies 35 providing a detectable signal. For improved assay performance,reaction chamber 34 can be maintained at a constant 37° C. - According to the present invention, the plastic fluidic cartridge need not be configured as a single-fluid delivery and analysis device.
FIG. 3 shows a plastic cartridge configured as a five fluid delivery and analysis device. Such a device can perform immunological assays, such as competitive immunoassay, immunosorbent immunoassay, immunometric immunoassay, sandwich immunoassay and indirect immunoassay, by providing immobilized antibodies inreaction chamber 46. Herereaction chamber 46 is not configured as a wide rectangular area, but a serpentine channel of dimensions similar to capillary dimension. This configuration provides more uniform flow through the reaction chamber at the expense of wasted space. For example, during immunoassays, a sample containing unknown concentrations of a plurality of antigens or antibodies is placed inreservoir 41. A wash buffer is placed inreservoir 42.Reservoir 43 remains empty to provide air purging. A substrate solution specific to the secondary antibody conjugate is placed inreservoir 44. The secondary antibody conjugate is placed inreservoir 45. Each reservoir is connected to apump structure 1′ similar to that ofFIG. 1 .Pump structures 1′ provide pumping fromreservoirs reaction chamber 46 to awaste reservoir 49. Asecondary reaction chamber 47 is provided for negative control and is isolated from the sample ofreservoir 41 bycheck valve 48. The protocol for performing immunoassays in this device is equivalent to that described previously for the single-fluid configuration with the distinct difference that each separated reagent is contained in a separate reservoir and pumped with a separate pump structure using a separate external linear actuator. First, an external linear actuator corresponding to a pump connected toreservoir 41 is repeatedly actuated until a sample fluid fillsreaction chamber 46. After a predetermined reaction time, the sample fluid is pumped to wastereservoir 49 using either a pump connected to samplereservoir 41 or a pump connected toair purge reservoir 43. Next the wash buffer is pumped intoreaction chamber 46 by repeatedly actuating the external actuator corresponding to a pump structure connected to washreservoir 42. The wash and/or air purge cycle can be repeated as necessary. A secondary antibody solution is then pumped intoreaction chamber 46 by repeatedly actuating the external linear actuator corresponding to a pump structure connected toreservoir 45. After a predetermined reaction time, the secondary antibody solution is excluded fromreaction chamber 46 either by a pump connected toreservoir 45 or a pump connected toair purge reservoir 43.Reaction chamber 46 is then washed as before. The substrate is pumped intoreaction chamber 46 by repeatedly actuating a linear actuator corresponding to a pump connected toreservoir 44. After a predetermined reaction time, the substrate is excluded fromreaction chamber 46 and replaced with wash buffer fromreservoir 42. Results of the immunoassay can then be confirmed by optical measurements throughupper substrate 21. - Furthermore, the reactions performed with the plastic fluidic cartridge of the invention need not be limited to reactions performed in stationary liquids.
FIG. 4 shows a plastic fluidic cartridge according to the invention, configured to provide continuous fluid motion throughreaction chamber 55. In this configuration,reservoirs FIG. 3 , but in this case the pump structures are connected to anintermediate circulation reservoir 56. For example,pump structure 57 is connected tocirculation reservoir 56 to provide continuous circulation of fluid fromcirculation reservoir 56 throughreaction chamber 55 and returning tocirculation reservoir 56. In this manner, a fluid can be circulated throughreaction chamber 55 without stopping. Such a fluid motion can provide better mixing, faster reactions times, and complete sample reaction with immobilized species inreaction chamber 55.Pump structure 58 is connected such that it provides pumping of fluids fromcirculation reservoir 56 to wastereservoir 54. Immunological assays similar to those described above can be performed in this device by immobilizing antibodies inreaction chamber 55 placing the sample containing unknown concentrations of antigens or antibodies in thecirculation reservoir 56, placing a solution of secondary antibody conjugate inreservoir 52, placing a substrate solution inreservoir 53, and placing a wash buffer inreservoir 51. The remaining protocol is identical to the above method with the addition of transferring fluids to and from thecirculation reservoir 56 and continuously circulating during all reaction times. - The system of the present invention can also be used to perform DNA hybridization analysis. Using the plastic cartridge of
FIG. 4 , a plurality of DNA probes are immobilized inreaction chamber 55. A sample containing one or more populations of fluorescently tagged, amplified DNA of unknown sequence is placed inreservoir 52. A first stringency wash buffer is placed inreservoir 51. A second stringency wash buffer is placed inreservoir 53.Reaction chamber 55 is maintained at a constant temperature of 52° C. The sample is transferred tocirculation reservoir 56 by repeatedly actuating a linear actuator corresponding to a pump structure connected toreservoir 52. The sample is then circulated throughreaction chamber 55 by repeatedly actuating a linear actuator corresponding to pumpstructure 57. The sample is circulated continuously for a predetermined hybridization time typically from 30 minutes to 2 hours. The sample is then excluded from thecirculation reservoir 56 andreaction chamber 55 by actuatingpump structures circulation reservoir 56 by repeatedly actuating the linear actuator corresponding to the pump structure connected toreservoir 51. The first stringency wash buffer is then circulated throughreaction chamber 55 in the same manner described above. After a predetermined wash time, the first stringency wash buffer is excluded fromreaction chamber 55 andcirculation reservoir 56 as described above. A second stringency wash buffer is then transferred tocirculation reservoir 56 and circulated throughreaction chamber 55 in a manner similar to that previously described. After the second wash buffer is excluded, the DNA hybridization results can be read by fluorescent imaging. - The invention being thus described, it will be obvious that the invention may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.
Claims (7)
1-11. (canceled)
12. A method of performing immunological assays of a fluid sample comprising a plurality of bio-molecules at unknown concentrations, comprising the steps of:
(a) placing said fluid sample into a sample reservoir defined in a fluidic cartridge;
(b) placing a wash buffer in a wash buffer reservoir defined in said fluidic cartridge;
(c) providing an air purge reservoir in said fluidic cartridge that remains empty to provide air purging;
(d) placing a secondary antibody conjugate in an antibody reservoir defined in said fluidic cartridge;
(e) placing a substrate solution specific to a secondary antibody conjugate in a substrate reservoir defined in said fluidic cartridge;
(f) pumping, by a pump structure, each of said fluid sample, said wash buffer, said substrate solution, and said secondary antibody conjugate to a waste reservoir through a first reaction chamber;
(g) providing a second reaction chamber for negative control and isolating said second reaction chamber from said fluid sample of said first reservoir by a check valve; and
(h) confirming results of immunoassay by optical measurements.
13. The method, as recited in claim 12 , wherein the step (f) further comprises the steps of:
(f-1) pumping said fluid sample from said sample reservoir to said first reaction chamber until said fluid sample at least partially fills said first reaction chamber;
(f-2) pumping said fluid sample from said first reaction chamber to said waste reservoir after a first predetermined reaction time;
(f-3) pumping, one or more times, said wash buffer from said wash buffer reservoir into said waste reservoir via said first reaction chamber;
(f-4) pumping said secondary antibody conjugate from said antibody reservoir into said first reaction chamber;
(f-5) pumping said secondary antibody conjugate from said first reaction chamber to said waste reservoir after a second predetermined reaction time;
(f-6) pumping said substrate solution from said substrate reservoir into said first reaction chamber; and
(f-7) pumping said substrate solution from said first reaction chamber to said waste reservoir after a third predetermined reaction time and at least partially filling said first reaction chamber with said wash buffer from said wash buffer reservoir.
14. The method, as recited in claim 13 , wherein said pump structure comprises a plurality of pumps and wherein said sample reservoir, said wash buffer reservoir, said air purge reservoir, said substrate reservoir, and said antibody reservoir is each connected to at least one pump in said plurality of pumps for pumping said fluid sample, said wash buffer, said substrate solution, and said secondary antibody conjugate through said first reaction chamber to said waste reservoir respectively.
15. The method, as recited in claim 13 , wherein said sample reservoir is a circulation reservoir and said pump structure is connected to said circulation reservoir to provide a continuous and repeated circulation of fluid, circulating from said circulation reservoir to said first reaction chamber and returning to said circulation reservoir, so that said fluid sample, said wash buffer, said substrate solution and said secondary antibody conjugate are capable of being circulated through said first reaction chamber without stopping.
16. The method, as recited in claims 13, 14 or 15, wherein said reservoir, said wash buffer reservoir, said air purge reservoir, said substrate reservoir, and said antibody reservoir is each connected to said first reaction chamber by one or more channels of capillary dimensions, wherein said fluidic cartridge includes a first substrate, a second substrate and an flexible intermediate interlayer sealedly interfaced between said first substrate and said second substrate to form therein said sample reservoir, said wash buffer reservoir, said air purge reservoir, said substrate reservoir, said antibody reservoir, said one or more channels, and said reaction chamber, and wherein said fluidic cartridge further provides a fluid flow controlling structure therein to restrict a flow of said fluid sample, said wash buffer, said substrate solution, and said second antibody conjugate through said first reaction chamber via said one or more channels in one direction only.
17. The method, as recited in claim 16 , wherein in said pumping step (f), said fluid sample, said wash buffer, said substrate solution, and said secondary antibody conjugate are respectively pumped by pumping actions in a plurality of pump chambers, wherein said plurality of pump chambers are defined in said fluidic cartridge and said pumping actions are provided by a plurality of linear actuators so as to pump said fluid sample, said wash buffer, said substrate solution, and said secondary antibody conjugate to flow from said sample reservoir, said wash buffer reservoir, said air purge reservoir, said substrate reservoir, and said antibody reservoir through said first reaction chamber via said one or more channels.
Priority Applications (1)
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US11/505,762 US20070020147A1 (en) | 2002-09-27 | 2006-08-16 | Miniaturized fluid delivery and analysis system |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
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TW091122431A TW590982B (en) | 2002-09-27 | 2002-09-27 | Micro-fluid driving device |
TW91122431 | 2002-09-27 | ||
US10/437,046 US7241421B2 (en) | 2002-09-27 | 2003-05-14 | Miniaturized fluid delivery and analysis system |
US11/505,762 US20070020147A1 (en) | 2002-09-27 | 2006-08-16 | Miniaturized fluid delivery and analysis system |
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US10/437,046 Division US7241421B2 (en) | 2002-09-27 | 2003-05-14 | Miniaturized fluid delivery and analysis system |
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US11/505,793 Active 2026-06-16 US8323887B2 (en) | 2002-09-27 | 2006-08-16 | Miniaturized fluid delivery and analysis system |
US11/505,762 Abandoned US20070020147A1 (en) | 2002-09-27 | 2006-08-16 | Miniaturized fluid delivery and analysis system |
US12/650,479 Abandoned US20100105065A1 (en) | 2002-09-27 | 2009-12-30 | Miniaturized Fluid Delivery and Analysis System |
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US11/504,303 Active - Reinstated 2024-11-04 US7666687B2 (en) | 2002-09-27 | 2006-08-15 | Miniaturized fluid delivery and analysis system |
US11/505,793 Active 2026-06-16 US8323887B2 (en) | 2002-09-27 | 2006-08-16 | Miniaturized fluid delivery and analysis system |
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Also Published As
Publication number | Publication date |
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US20070031287A1 (en) | 2007-02-08 |
US8323887B2 (en) | 2012-12-04 |
US20100105065A1 (en) | 2010-04-29 |
CN100394184C (en) | 2008-06-11 |
TW590982B (en) | 2004-06-11 |
US20040063217A1 (en) | 2004-04-01 |
US20070020148A1 (en) | 2007-01-25 |
US7241421B2 (en) | 2007-07-10 |
US7666687B2 (en) | 2010-02-23 |
CN1548957A (en) | 2004-11-24 |
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