Device and method for microfluidic control of a first fluid in contact with a second fluid, wherein the first and second fluids are immiscible

• US 20070068573A1
• Filed: 08/22/2006
• Published: 03/29/2007
• Est. Priority Date: 08/22/2005
• Status: Abandoned Application

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Cox, David, Oldham, Mark, Wiyatno, Willy, Boege, Steven, Nordman, Eric, Lee, Linda, Sagatelyan, Dmitry, Reel, Richard, Woo, Sam
• Application Number:
  US11/508,756
• Publication Number:
  US 20070068573A1
• Time in Patent Office: Days
• US Class Current: 137/1
• CPC Class Codes: B01J 2219/00353 Pumps
• CPC Class Codes: B01J 2219/0036 Nozzles
• CPC Class Codes: B01J 2219/00364 Pipettes
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00657 One-dimensional arrays
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/0293 for liquids
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502784 : specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12Q 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6869 : Methods for sequencing
• CPC Class Codes: C12Q 1/6874 : involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2535/101 : Sanger sequencing method, i.e. oligonucleotide sequencing using primer elongation and dideoxynucleotides as chain terminators
• CPC Class Codes: F15C 5/00 : Manufacture of fluid circuit elements; Manufacture of assemblages of such elements integrated circuits
• CPC Class Codes: F16K 2099/0084 : Chemistry or biology, e.g. "lab-on-a-chip" technology
• CPC Class Codes: F16K 99/0001 : Microvalves microdevices B81B1/00; manufacture or treatment of devices or systems in or on a substrate B81C1/00; microfluidic structures B01L3/5027; micropumps F04B19/006
• CPC Class Codes: F16K 99/0011 : Gate valves or sliding valves
• CPC Class Codes: F16K 99/0013 : Rotary valves
• CPC Class Codes: G01N 1/14 : Suction devices, e.g. pumps; Ejector devices
• CPC Class Codes: G01N 27/44743 : Introducing samples
• CPC Class Codes: G01N 27/44769 : Continuous electrophoresis, i.e. the sample being continuously introduced, e.g. free flow electrophoresis [FFE]
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: Y10T 137/0318 : Processes
• CPC Class Codes: Y10T 137/4259 : With separate material addition
• CPC Class Codes: Y10T 137/85978 : With pump
• CPC Class Codes: Y10T 137/85986 : Pumped fluid control
• CPC Class Codes: Y10T 137/86863 : Rotary valve unit
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

Various embodiments described in the application relate to an apparatus, system, and method for fluidically controlling, within a conduit, discrete volumes of one or more fluids that are immiscible with a second fluid. The discrete volumes can be used for biochemical or molecular biology procedures involving small volumes, for example, microliter-sized volumes, nanoliter-sized volumes, or smaller. The system can comprise combinations of detectors, controllers, valves, and fluid supply units to control the spatial size, location and/or fluidic composition of discrete volumes separated from one another by a fluid that is immiscible with the fluid(s) of the discrete volumes, for example, aqueous immiscible-fluid-discrete volumes separated by an oil.

Claims

1. A method of splitting a discrete volume of a liquid contained in a conduit into two or more smaller discrete volumes of the liquid, the method comprising:

connecting to a first displacement pump a first conduit for receiving a first, smaller discrete volume of a first liquid;

connecting to a second displacement pump a second conduit for receiving a second, smaller discrete volume of the first liquid;

supplying to a junction of at least the first and second conduits a discrete volume of a first liquid in contact with a second liquid with which the first liquid is immiscible; and

moving the first and second displacement pumps in a first direction at the same time, thereby splitting the discrete volume of the first liquid into at least the first and second smaller discrete volumes of the first liquid;

wherein the ratio of the volume of the first smaller discrete volume of the first liquid to the volume of the second smaller discrete volume of the first liquid is based on the ratio of the volume drawn by the first displacement pump to the volume drawn by the second displacement pump.

2. The method of claim 1, wherein moving the first and second displacement pumps in a first direction at the same time comprises moving a linkage attached to at least the first and second displacement pumps.

3. The method of claim 2, further comprising:

connecting the common port of a first three-way valve to the first displacement pump;

connecting the first of two input/output ports of the first three-way valve to the first conduit;

connecting the second of two input/output ports of the first three-way valve to a third conduit;

connecting the common port of a second three-way valve to the output of the second displacement pump;

connecting the first of two input/output ports of the second three-way valve to the second conduit;

connecting the second of two input/output ports of the second three-way valve to a fourth conduit;

after moving the linkage in a first direction, moving the linkage in a second direction opposite the first direction, thereby displacing a first and second volume of the second liquid into the third and fourth conduits, respectively.

4. The method of claim 2, wherein the supplying to a junction comprises:

detecting the speed of the discrete volume of the first fluid, which is flowing in a third conduit connected to the junction of at least the first and second conduits, at a known distance from the junction; and

stopping the flow of the discrete volume of the first fluid at the junction.

5. An apparatus comprising:

a first displacement pump having a first cross-sectional area;

a first conduit for receiving a first, smaller discrete volume of a first liquid connected to the first displacement pump;

a second displacement pump having a second cross-sectional area;

a second conduit for receiving a second, smaller discrete volume of the first liquid connected to the second displacement pump;

a third conduit containing a discrete volume of a first liquid in a second liquid with which the first liquid is immiscible to, a junction of at least the first and second conduits; and

a linkage attached to at least the first and second displacement pumps, wherein when the linkage is moved in a first direction, the discrete volume of the first liquid splits into at least the first and second smaller discrete volumes of the first liquid, and the ratio of the volume of the first smaller discrete volume of the first liquid to the volume of the second smaller discrete volume of the first liquid is based on the ratio of the cross-sectional area of the first displacement pump to the cross-sectional area of the second displacement pump.

6. The apparatus of claim 5, further comprising:

a first three-way valve having a common port, and at least two input/output ports, wherein the common port of the first three-way valve is connected to the output of the first displacement pump, the first of two input/output ports of the first three-way valve is connected to the first conduit, and the second of two input/output ports of the first three-way valve is connected to a fourth conduit;

a second three-way valve having a common port and at least two input/output ports, wherein the common port of a second three-way valve is connected to the output of the second displacement pump, the first of two input/output ports of the second three-way valve is connected to the second conduit, and the second of two input/output ports of the second three-way valve is connected to a fifth conduit;

wherein when the linkage is moved in a second direction opposite the first direction, the first displacement pump displaces a first volume of the second liquid into the fourth conduit and the second displacement pump displaces a second volume of the second liquid into the fifth conduit.

7. The apparatus of claim 5, further comprising a detector adjacent the third conduit and operatively coupled to the motor.

8. The apparatus of claim 5, wherein at least one of the first and second three-way valves is a rotary three-way valve.

9. A method of adding two or more different miscible liquids in predetermined volumetric combination to an existing immiscible-fluid, discrete volume, the method comprising:

connecting to a first displacement pump a first conduit for supplying a first liquid;

connecting to a second displacement pump a second conduit for supplying a second liquid miscible with the first liquid;

connecting to a junction of the first and second conduits a third conduit for receiving the predetermined volumetric combination of the first liquid and second liquid, moving a linkage attached to the first and second displacement pumps in a first direction, thereby combining the first and second liquid in the predetermined volumetric relationship;

thereby creating an addition liquid;

supplying the addition liquid to a fourth conduit containing an immiscible-fluid, discrete volume of a fourth liquid, miscible with the addition liquid; and

adding a volume of the addition liquid to the immiscible-fluid, discrete volume.

10. The method of claim 9, further comprising:

connecting the common port of a first three-way valve to the first displacement pump;

connecting the first of two input/output ports of the first three-way valve to the first conduit;

connecting the second of two input/output ports of the first three-way valve to a fourth conduit;

connecting the common port of a second three-way valve to the second displacement pump;

connecting the first of two input/output ports of the second three-way valve to the second conduit;

connecting the second of two input/output ports of the second three-way valve to a fifth conduit;

before moving the linkage in a first direction, moving the linkage in a second direction opposite the first direction, thereby withdrawing a first and second volume of the second liquid from the fourth and fifth conduits, respectively.

11. A method of adding a first liquid in predetermined volumetric relationship to a second liquid with which it is immiscible, the method comprising:

connecting to a first displacement pump a first conduit for supplying a first liquid;

connecting to a second displacement pump a second conduit for supplying a second liquid immiscible with the first liquid;

connecting to a junction of the first and second conduits a third conduit for receiving immiscible-fluid, discrete volumes of at least one of the first liquid and second liquid,

moving a linkage attached to the first and second displacement pumps in a first direction, thereby generating immiscible-fluid, discrete volumes of at least the first liquid in the third conduit;

thereby creating a first set of immiscible-fluid, discrete volumes of the first liquid.

12. A method of adding a first liquid to a third liquid with which it is miscible, the method comprising

connecting to a first displacement pump a first conduit for supplying a first immiscible-fluid, discrete volume of a first liquid in contact with a second fluid with which the first liquid is immiscible;

connecting to a second displacement pump a second conduit for supplying a second immiscible-fluid, discrete volume of a third liquid in contact with the second fluid which the third liquid is immiscible;

connecting to a junction of the first and second conduits a third conduit for receiving a larger, immiscible-fluid, discrete volume of the first and third liquids in contact with the second fluid, with which the first and third liquids are each immiscible,

positioning the first immiscible-fluid, discrete volume in the first conduit and the second immiscible-fluid, discrete volume in the second conduit;

moving a linkage attached to the first and second displacement pumps in a first direction, thereby contacting the immiscible-fluid, discrete volume of first liquid with the immiscible-fluid, discrete volume of the third liquid to form a larger immiscible-fluid, discrete volume of the first and third liquids.

13. The method of claim 12, further comprising:

connecting the common port of a first three-way valve to the first displacement pump;

connecting the first of two input/output ports of the first three-way valve to the first conduit;

connecting the second of two input/output ports of the first three-way valve to a fourth conduit;

connecting the common port of a second three-way valve to the second displacement pump;

connecting the first of two input/output ports of the second three-way valve to the second conduit;

connecting the second of two input/output ports of the second three-way valve to a fifth conduit;

before moving the linkage in a first direction, moving the linkage in a second direction opposite the first direction, thereby withdrawing the first immiscible-fluid-discrete volume and the second immiscible-fluid-discrete-volume from the fourth and fifth conduits, respectively.

14. An apparatus comprising:

a conduit containing discrete volumes of aqueous liquid in contact with an immiscible, non-aqueous liquid;

an expandable and contractible reservoir in fluid communication with the conduit in at least one location and containing a volume of the immiscible, non-aqueous liquid;

a diaphragm coupled to the reservoir;

an enclosure coupled to the diaphragm, such that the diaphragm is between the enclosure and the reservoir;

wherein when the space between the enclosure and the diaphragm is positively pressurized, the fluid in the conduit is pressurized.

15. The apparatus of claim 14, wherein the conduit is in further fluid communication with the reservoir at a second location.

16. The apparatus of claim 15, wherein between the first and second locations, the conduit is adjacent a thermal cycler.

17. The apparatus of claim 15, wherein between the first and second locations, the conduit is adjacent at least one of the following:

a heating unit, cooling unit, isothermal zone, and a thermal cycler.

18. An apparatus comprising:

a stator having at least three through-holes from outer diameter to inner diameter;

a mating rotor having at least two through-holes, first, second, and third tubes, each coupled to one of the three through-holes of the stator, wherein at least one of the first, second, and third tubes contains one or more immiscible-fluid, discrete volumes;

wherein, when the stator and mating rotor are in a first relative position, a first of the at least two through-holes in the mating rotor aligns with two of the at least three through-holes in the stator, and when the stator and mating rotor are in a second relative position, a second of the at least two through-holes in the mating rotor is in fluid communication with one of the previous two of the at least three through-holes and the remaining third through-hole in the stator.

19. The apparatus of claim 18, wherein at least the mating rotor and the first and second conduits are made from the same type of material.

20. An apparatus for preventing flow of discrete-volumes of a first fluid in contact with a second fluid with which the first fluid is immiscible, the apparatus comprising:

a first member connected to a first tube for supplying one or more discrete volumes of a first fluid in contact with a second fluid with which the first fluid is immiscible;

a second member having a through-hole for receiving discrete volumes of the first fluid from the first tube in a first position,

wherein when the second member is in a second position with respect to the first member, discrete volumes of the first fluid cannot flow through the through-hole in the second member.

21. The apparatus of claim 20, wherein the second member is rotatable between the first and second position.

22. The apparatus of claim 20, wherein the second member is translatable between the first and second position.

23. An apparatus for preventing flow of discrete-volumes of a first fluid in contact with a second fluid with which the first fluid is immiscible, the apparatus comprising:

an immiscible-fluid-discrete-volume tube;

a first pinching member having a radiused surface in at least tangential contact with the immiscible-fluid-discrete-volume tube;

a second pinching member in a first position with respect to the immiscible-fluid-discrete-volume tube, wherein a section of the tube is pinched between the radiused surface of the first pinching member and the second pinching member, such that discrete volumes of the first fluid cannot flow past the compressed section.

24. The apparatus of claim 23 further comprising a solenoid in contact with one of the first and second pinching member.

25. A method of splitting a discrete volume of a first fluid in contact with a second fluid with which it is immiscible, the method comprising:

positioning a first part of an immiscible-fluid-discrete-volume in a through-hole of a first member that is in a first position, wherein a through-hole in the first member is aligned with an immiscible-fluid-discrete-volume supply tube;

moving the first member to a second position, wherein the through-hole in the first member is aligned with a spacing-fluid supply tube, thereby turning the first part of an immiscible-fluid-discrete-volume into a first-split-part, immiscible-fluid, discrete volume in the through-hole;

moving the first-split-part, immiscible-fluid, discrete volume into a first-split-part tube that is aligned with the through-hole in the second position;

moving the first member back to the first position, wherein the through-hole is aligned with the immiscible-fluid-discrete-volume supply.

26. The method of claim 25, wherein positioning a first part comprises:

detecting the speed of an immiscible-fluid, discrete volume in the immiscible-fluid-discrete-volume supply conduit at a known distance from the first member;

stopping the force moving the immiscible-fluid, discrete volume; and

detecting a known point of the immiscible-fluid, discrete volume in the first member at a desired distance from an opening of the through-hole in the first member.

27. A method of removing gas phase bubbles from spacing fluid between discrete volumes of a first fluid which is immiscible with the spacing fluid, the method comprising:

flowing a stream of fluid comprising at least two immiscible-fluid-discrete-volumes separated by spacing fluid and a gas phase bubble therein in a first conduit in a first direction toward a junction of the first conduit, a second conduit and at least a third conduit;

flowing spacing fluid in the second conduit away from the junction in a vertical direction;

flowing at least one of the at least two immiscible-fluid-discrete-volumes separated by spacing fluid into the third conduit, the longitudinal axis of which forms an angle in the junction of greater than zero and less than 180 degrees with the longitudinal axis of the second conduit at the junction;

flowing the gas phase bubble into the junction, wherein the gas phase bubble transfers, due to a smaller specific gravity than the spacing fluid, to the spacing fluid flowing away from the junction in a vertical direction, thereby removing the gas phase bubble from the spacing fluid between the two of the at least two immiscible-fluid-discrete-volumes.

28. The method of claim 27, wherein the angle in the junction between the longitudinal axis of the third conduit and the longitudinal axis of the second conduit is approximately 90 degrees.

29. The method of claim 27 further comprising:

flowing a stream of spacing fluid in a fourth conduit toward the junction, wherein the longitudinal axis of the fourth conduit forms an angle in the junction with the longitudinal axis of the second conduit of approximately 90 degrees.

30. A method comprising:

alternately introducing a first fluid and a second fluid, that is immiscible with the first fluid, into a conduit, to form a set of immiscible discrete volumes of the second fluid, each immiscible discrete volume of the set being separated from one or more other immiscible discrete volumes of the set by the first fluid, the set comprising a first end and a second end;

moving the set of immiscible discrete volumes in a first direction by withdrawing from the conduit, some of the first fluid from the first end of the set; and

moving the set in the first direction by adding to the conduit, more first fluid at the second end of the set.

31. The method of claim 30, wherein the first fluid comprises an oil and the second fluid comprises an aqueous liquid that is immiscible in the oil.

32. The method of claim 30, further comprising moving the set past a valve in the conduit and closing the valve before moving the set in the first direction by adding to the conduit more first fluid at the second end of the set.

33. The method of claim 32, wherein the closing the valve comprises rotating a rotary valve.

34. The method of claim 32, wherein the closing the valve comprising actuating a linear, sliding valve.