Device and method for making discrete volumes of a first fluid in contact with a second fluid, which are immiscible with each other

• US 20070062583A1
• Filed: 08/22/2006
• Published: 03/22/2007
• Est. Priority Date: 08/22/2005
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Lehto, Dennis, Cox, David, Wojtowicz, Janusz, Oldham, Mark, Greiner, Douglas, Wiyatno, Willy, Nurse, James, Lee, Linda, Reel, Richard, Woo, Sam
• Granted Patent: US 7,955,864 B2
• Time in Patent Office: Days
• US Class Current: 137/565.10
• 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 generating, 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 an apparatus comprising at least one conduit operatively connected to one or more pumps for providing 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. An apparatus comprising:

a first conduit having an outer perimeter and a length;

a second conduit having an inner perimeter, wherein at least a portion of the length of the first conduit is inside of the second conduit, thereby defining a space between the outer perimeter of the first conduit and the inner perimeter of the second conduit;

a first pump operatively connected to the first conduit, wherein the first pump is configured to flow fluids through the first conduit in a first direction; and

a second pump operatively connected to the second conduit, wherein the second pump is configured to flow a second fluid through the second conduit in a second direction that is opposite the first direction.

2. The apparatus of claim 1, further comprising a control unit configured to synchronize actuation of the first pump and the second pump.

3. The apparatus of claim 1, further comprising a conduit positioner configured to axially move one of the first conduit and the second conduit with respect to the other.

4. The apparatus of claim 3, further comprising a control unit configured to synchronize actuation of the first pump, the second pump, and the conduit positioner.

5. The apparatus of claim 1, wherein the first conduit has an end surface, the second conduit has an end surface, and the end surface of the first conduit is beyond the end surface of the second conduit.

6. The apparatus of claim 1, wherein the second conduit comprises a block having a through-hole.

7. The apparatus of claim 6, wherein the first conduit comprises an end surface and the end surface is disposed within the through-hole of the block.

8. The apparatus of claim 6, wherein the block further comprises a passageway formed therein;

the passageway having a first end at the through-hole and extending away from the through-hole.

9. The apparatus of claim 8, further comprising a third pump operatively connected to the passageway and configured to draw fluid from the through-hole into the passageway.

10. The apparatus of claim 1, wherein the first conduit has a first inner diameter, the first inner diameter is from about 10 microns to about 2000 microns, the second conduit has a second inner diameter, and the second inner diameter is from about 20 microns to about 5000 microns.

11. The apparatus of claim 1, wherein the first conduit comprises a tube.

12. A system comprising the apparatus of claim 3, and a sample liquid disposed in a sample container, wherein the conduit positioner is configured to move a tip of the first conduit into the sample container and into contact with the sample liquid.

13. A system for aspirating liquids, comprising:

at least one differential pressure source;

an aspirating tube in communication with the at least one differential pressure source and comprising an intake tip;

at least one fluid container in communication with the intake tip, the at least one fluid container containing at least a first fluid and a second fluid, the first fluid and the second fluid being immiscible with each other; and

at least one intake tip positioning unit configured to alternately aspirate the first fluid and the second fluid by raising and lowering one of the at least one fluid container and the intake tip relative to the other and using differential pressure delivered by the at least one differential pressure source.

14. A system according to claim 13, wherein at least one of the first fluid and the second fluid are covered with a covering liquid layer.

15. A system according to claim 13, wherein the at least one fluid container comprises a set of wells.

16. A system according to claim 13, wherein the at least one fluid container comprises a first well containing therein both the first fluid and the second fluid in separate layers.

17. A method comprising:

disposing an intake tip of an aspirating conduit in a fluid container comprising at least first and second fluids that are immiscible with one another and form layers in the fluid container;

aspirating the first fluid through the intake tip and into the aspirating conduit;

moving one of the intake tip and the fluid container up or down relative to the other until the intake tip is immersed in the second fluid in the fluid container; and

aspirating the second fluid through the intake tip and into the aspirating conduit.

18. The method of claim 17, further comprising then moving the one of the intake tip and the fluid container back until the intake tip is immersed in the first fluid and then aspirating the first fluid into the aspirating conduit.

19. The method of claim 18, further comprising then moving the one of the intake tip and the fluid container back until the intake tip is immersed in the second fluid and aspirating the second fluid into the aspirating conduit.

20. A method comprising:

flowing a first fluid into a through-hole of a slider housed in a slider housing and positioned at a first position;

shifting the slider from the first position to a second position relative to the slider housing, to align the through-hole of the slider with an output conduit containing a second fluid that is immiscible with the first fluid; and

moving the first fluid from inside the through-hole out of the through-hole and into the output conduit, with a source of additional second fluid to form a discrete volume of the first fluid in contact with the second fluid.

21. The method of claim 20, further comprising moving some of the second fluid through the through-hole in the slider and into the output conduit adjacent the first fluid.

22. The method of claim 21, further comprising:

shifting the slider back to the first position;

moving the second fluid from inside the through-hole out of the through-hole, with the first fluid that is immiscible with the second fluid, filling the through-hole with a second portion of the first fluid;

shifting the slider from the first position back to the second position, to again align the through-hole of the slider with the output conduit; and

moving the second portion of the first fluid from inside the through-hole out of the through-hole and into the output conduit, with more of the second fluid, to form a second discrete-volume of the first fluid in contact with the second fluid.

23. The method of claim 22, wherein the volume of the first discrete-volume and the volume of the second discrete-volume are the same volume.

24. A system comprising:

a housing;

a slider arranged in the housing for sliding movement therein between at least a first position and a second position, the slider comprising a through hole;

a first fluid supply conduit operatively connected to a first end of the through-hole when the slider is in the first position;

a waste conduit operatively connected to a second, opposite end of the through-hole when the slider is in the first position;

a second fluid supply conduit operatively connected to the first end of the through-hole when the slider is in the second position;

an immiscible-fluid-discrete-volume-forming conduit operatively connected to the second, opposite end of the through-hole when the slider is in the second position;

a supply of a first fluid operatively connected to the first fluid supply conduit; and

a supply of a second fluid operatively connected to the second fluid supply conduit, wherein the second fluid and the first fluid are immiscible with respect to one another.

25. A method comprising:

merging together at an junction of a first pair of conduits and a second pair of conduits a first fluid and a second fluid, the first fluid comprising a spacing fluid and the second fluid comprising an immiscible-discrete-volume-forming fluid that is immiscible with the first fluid, such that a set of immiscible-fluid-discrete-volumes of the second fluid are formed that are spaced apart from one another by the first fluid, wherein at least one of the first fluid, the second fluid, and the set of immiscible-fluid-discrete-volumes flows through a rotary valve comprising a stator and a rotor, the first pair of conduits comprises a first conduit and a second conduit that each pass through the stator, the second pair of conduits comprises a third conduit and a fourth conduit that each pass through the stator, and the rotor comprises a through-hole that, in a first position, fluidly communicates the first conduit with the second conduit, and in a second position fluidly communicates the third conduit with the fourth conduit, and the method further comprises rotating the rotor from the first position to the second position.

26. The method of claim 25, wherein at least a portion of each of the first conduit, the second conduit, and the third conduit is disposed in the rotor, and the junction comprises an junction of the three portions in the rotor.

27. The method of claim 25, wherein the rotor comprises at least two independent, non-intersecting through-holes formed therein, one of the at least two through-holes is in fluid communication with the first conduit, and another of the at least two through-holes is in fluid communication with the second conduit.

28. A system comprising:

a rotary valve comprising a stator and a rotor;

a first conduit in operatively connected to the rotary valve;

a second conduit operatively connected to the rotary valve;

an junction of the first conduit and the second conduit in the rotor of the rotary valve;

a first fluid comprising a spacing fluid in fluid communication with the first conduit;

a second fluid comprising an immiscible-discrete-volume-forming fluid, that is immiscible with the first fluid, in fluid communication with the second conduit;

a third conduit operatively connected to the rotary valve and in fluid communication with the intersection; and

a fourth conduit operatively connected to the rotary valve and in fluid communication with the intersection, wherein the rotor comprises a through-hole that, in a first position, fluidly communicates the first conduit with the second conduit, and in a second position fluidly communicates the third conduit with the fourth conduit.

29. A system comprising:

an immiscible-fluid-discrete-volume-forming conduit comprising an intake end;

an electro-wetting device comprising one or more electro-wetting pathways for transporting one or more droplets, and an output site along at least one of the one or more electro-wetting pathways; and

a positioning unit for positioning the intake tip adjacent or at the output site.

30. The system of claim 29, further comprising a negative pressure source operatively connected to the immiscible-fluid-discrete-volume-forming conduit.

31. A method comprising:

transporting a first droplet of a first fluid along an electro-wetting pathway of an electro-wetting device;

merging the first droplet with a second droplet of a second fluid that is miscible with the first fluid, to form a third droplet; and

drawing the third droplet into an immiscible-fluid-discrete-volume-forming conduit.

32. The method of claim 31, further comprising, prior to the drawing step, first drawing a third fluid into the immiscible-fluid-discrete-volume-forming conduit, wherein the third fluid is immiscible with the immiscible-fluid-discrete-volume.

33. The method of claim 32, further comprising drawing more of the third fluid into the immiscible-fluid-discrete-volume-forming conduit after drawing the immiscible-fluid-discrete-volume into the immiscible-fluid-discrete-volume-forming conduit.

34. A device comprising:

a substrate and an elastically deformable bottom cover, the substrate comprising

a bottom wall having a central axis of rotation and a lower surface,

an annular wall extending upward from the bottom wall and defining a central reservoir radially inward with respect to the annular wall,

a plurality of through-holes in the bottom wall in the central reservoir,

a plurality of radial reservoirs formed in the substrate and disposed radially outward with respect to the annular wall, each radial reservoir comprising at least one sidewall and a bottom, and

a plurality of through-holes, at least one in the bottom of each radial reservoir;

wherein the elastically deformable bottom cover is attached to the lower surface of the bottom wall and spaced-apart from portions of the lower surface of the bottom wall such that a respective radial fluid channel is provided between each through-hole in the central reservoir and a respective through-hole of the plurality of through-holes in the radial reservoirs.

35. The device of claim 34, wherein the central reservoir contains a first fluid, and at least a first radial reservoir of the plurality of radial reservoirs contains a second fluid that is immiscible in the first fluid.

36. The device of claim 35, wherein the elastically deformable bottom cover is configured to create positive pressure in the radial fluid channel between the first radial reservoir and the respective through-hole in the central reservoir when the elastically deformable bottom cover is pushed upwardly adjacent the radial fluid channel such that second fluid in the radial fluid channel is forced through the through-hole in the central reservoir and into the central reservoir.

37. A system comprising:

the device of claim 34;

a rotatable support comprising a holder for holding the device;

a drive unit for rotating the rotatable support, while holding the device, about the central axis of rotation; and

a plunger configured to press against the bottom cover.

38. The system of claim 37, further comprising a negative pressure source and a conduit, the conduit being operatively connected to the negative pressure source and comprising an intake tip positioned in the central reservoir.

39. A method comprising:

forming a first droplet of a second fluid in a first fluid, wherein the first fluid and the second fluid are immiscible with respect to one another and have different densities;

moving at least one of the first droplet and an intake tip of a conduit relative to one another such that the first droplet is disposed adjacent the intake tip; and

drawing the first droplet and an amount of the first fluid through the intake tip and into the conduit.

40. The method of claim 39, further comprising:

forming in the first fluid a second droplet of a third fluid that is miscible with the second fluid; and

merging the second droplet with the first droplet, thereby forming a merged droplet of first and third fluid, before the drawing step, wherein the drawing comprises drawing the merged droplet through the intake tip and into the conduit.

41. The method of claim 39, wherein the first fluid is disposed in a container having a bottom, and the second fluid is less dense than the first fluid, and further comprising

ejecting a volume of the second fluid from a through-hole in the bottom, thereby forming the first droplet, which rises in the first fluid.

42. The method of claim 39, wherein the second fluid has a greater density than the first fluid, the droplet of the second fluid is formed on a film, and the method further comprises submersing the film in the first fluid.

43. A method comprising:

providing a device, the device comprising a substrate, the substrate comprising

a bottom wall having a central axis of rotation and a lower surface,

an annular wall extending upward from the bottom wall and defining a central reservoir radially inward with respect to the annular wall,

a first through-hole extending through the bottom wall in the central reservoir,

a radial reservoir formed in the substrate and disposed radially outward with respect to the annular wall, the radial reservoir comprising at least one sidewall and a bottom, and

a second through-hole extending through the bottom of the radial reservoir;

the device further comprising an elastically deformable bottom cover attached to the lower surface of the bottom wall and spaced-apart from a portion of the lower surface of the bottom wall such that a respective radial fluid channel is provided between the first through-hole and the second through hole, wherein the central reservoir contains a first fluid, the radial fluid channel comprises a second fluid, the second fluid is less dense than the first fluid, the first fluid and the second fluid are immiscible with respect to one another, and the method comprises

forcing the elastically deformable bottom cover upwardly toward the lower surface of the bottom wall to create positive pressure in the respective radial fluid channel that forces a droplet of the second fluid to exit the first through-hole and enter the central reservoir.

44. The method of claim 43, further comprising drawing the droplet of the second fluid and surrounding first fluid into an intake tip of a conduit.

45. A system comprising:

an immiscible-fluid-discrete-volume-forming conduit; and

a magnetohydrodynamic pump device comprising one or more magnetohydrodynamic pumps configured to transport one or more immiscible-fluid-discrete-volumes to the immiscible-fluid-discrete-volume-forming conduit;

a supply of a first fluid operatively connected to the one or more magnetohydrodynamic pumps; and

a supply of a second fluid operatively connected to the one or more magnetohydrodynamic pumps, wherein the second fluid is immiscible with the first fluid.

46. The system of claim 45, further comprising a negative pressure source operatively connected to the immiscible-fluid-discrete-volume-forming conduit.

47. A method comprising:

actuating a magnetohydrodynamic pump to transport a first droplet of a first fluid along a pathway of a device;

merging the first droplet with a second droplet of a second fluid that is miscible with the first fluid, to form a third droplet; and

introducing the droplet into an immiscible-fluid-discrete-volume-forming conduit.

48. The method of claim 47, further comprising, moving a third fluid into the immiscible-fluid-discrete-volume-forming conduit, wherein the third fluid is immiscible with the third droplet.

49. The method of claim 47, wherein the introducing comprises actuating a magnetohydrodynamic pump.

50. A conduit rinsing system comprising:

a tubular conduit including a tip and an outer surface;

a cleaning vessel comprising a top, a bottom, an outer annular wall, an inner annular wall having a top rim and an inner surface, a space provided between the outer annular wall and the inner annular wall, and at least one port formed in the bottom and communicating with the space, wherein the tip is disposed in the cleaning vessel between the top and the bottom; and

a closure flap disposed between the top of the cleaning vessel and the top rim of the inner annular wall, the closure flap being spaced from the top rim, wherein a rinse space is provided between the inner surface of the inner annular wall and the outer surface of the conduit, and an opening is provided in the bottom and in fluid communication with the rinse space.

51. The conduit rinsing system of claim 50, further comprising a supply of at least one cleaning fluid operatively connected to the at least one port.

52. A method comprising:

applying a negative pressure to a conduit system comprising an intake tip;

contacting the intake tip with a first fluid and a second fluid that is immiscible with the first fluid, while applying the negative pressure, to draw the first fluid and the second fluid into the conduit system and form a set of discrete volumes of the first fluid spaced apart from one another by the second fluid, the set moving in a first direction in the conduit system; and

thereafter, applying a positive pressure to the conduit system to push the set of discrete volumes in the conduit system.

53. The method of claim 52, wherein the applying a positive pressure causes the set to move in the first direction.

54. The method of claim 52, wherein the applying a positive pressure causes the set to move in the conduit system a second direction that is opposite the first direction.

55. The method of claim 52, wherein the negative pressure is applied to the conduit system until the set moves past a first diverter and the method comprises then changing a position of the diverter before applying the positive pressure.

56. The method of claim 52, wherein the negative pressure is applied to the conduit system until the set moves past a valve and a port, and the method comprises then closing the valve, and wherein the applying positive pressure comprises applying a positive pressure through the port.

57. The method of claim 52, comprising applying the negative pressure with a reversible pump, and reversing the action of the reversible pump to apply the positive pressure.

58. The method of claim 52, wherein the contacting further comprises applying an electro-wetting force to move at least one of the first fluid and the second fluid to a location adjacent the intake tip.