SYSTEM FOR FORMING AN ARRAY OF EMULSIONS

• US 20110092392A1
• Filed: 12/07/2010
• Published: 04/21/2011
• Est. Priority Date: 09/23/2008
• Status: Active Grant

• Current Assignee: Lawrence Livermore National Security LLC (Government of the United States of America)
• Original Assignee: QuantaLife, Inc. (Bio-Rad Laboratories Incorporated)
• Inventors: Makarewicz, Anthony Joseph JR., Colston, Billy Wayne JR., Milanovich, Fred Paul, Ness, Kevin Dean, Masquelier, Donald Arthur, Hindson, Benjamin Joseph
• Granted Patent: US 9,216,392 B2
• Time in Patent Office: Days
• US Class Current: 506/27
• CPC Class Codes: B01F 2101/23 Mixing of laboratory samples e.g. in preparation of analysing or testing properties of materials
• CPC Class Codes: B01F 23/41 Emulsifying
• CPC Class Codes: B01F 23/4143 Microemulsions
• CPC Class Codes: B01F 23/4145 Emulsions of oils, e.g. fuel, and water
• CPC Class Codes: B01F 33/3011 using a sheathing stream of a fluid surrounding a central stream of a different fluid, e.g. for reducing the cross-section of the central stream or to produce droplets from the central stream
• CPC Class Codes: B01F 35/10 Maintenance of mixers
• CPC Class Codes: B01F 35/165 Making mixers or parts thereof
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/0689 Sealing
• 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 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/041 Connecting closures to device or container
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0858 Side walls
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2400/0478 pistons
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/049 : vacuum
• CPC Class Codes: B01L 2400/0622 : distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 3/0241 : Drop counters; Drop formers making arrays for combinatorial libraries B01J19/0046; automation of dispensing for analysis G01N35/10
• 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: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: B29C 2045/0079 : applying a coating or covering
• CPC Class Codes: B29C 45/0053 : combined with a final operation, e.g. shaping injection-compression moulding B29C45/561
• CPC Class Codes: B29C 45/006 : Joining parts moulded in separate cavities
• CPC Class Codes: B29L 2031/752 : Measuring equipment
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 2021/6439 : with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/3563 : for analysing solids; Preparation of samples therefor
• CPC Class Codes: G01N 21/49 : within a body or fluid
• CPC Class Codes: G01N 21/6428 : Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6486 : Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: Y02A 90/10 : Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

Abstract

System, including apparatus and methods, for forming an array of emulsions. The system may comprise a plate including an array of emulsion production units. Each unit may include at least one first input well, a second input well, and an output well connected to the first and second input wells by channels that form a droplet generator. The system also may comprise a vacuum or pressure source configured to be connected operatively to wells of the plate to form a pressure drop between the input wells and the output well of each unit that is capable of driving a first fluid and a second fluid from respective first and second input wells of such unit and through the droplet generator, for collection as an emulsion in the output well of such unit.

Claims

1. A system for forming an array of emulsions, comprising:

a plate including an array of emulsion production units, each unit including

at least one first input well,

a second input well, and

an output well connected to the first and second input wells by channels that form a droplet generator; and

a vacuum or pressure source configured to be connected operatively to wells of the plate to form a pressure drop between the input wells and the output well of each unit that is capable of driving a first fluid and a second fluid from respective first and second input wells of such unit and through the droplet generator, for collection as an emulsion in the output well of such unit.

2. The system of claim 1, wherein the vacuum or pressure source controls air pressure in wells to which the source is configured to be connected operatively.

3. The system of claim 1, further comprising an instrument including the vacuum or pressure source and a manifold, wherein the vacuum or pressure source is configured to be connected operatively to wells of the plate via the manifold.

4. The system of claim 3, further comprising one or more gaskets that engage the plate and the manifold to form a seal between the manifold and the wells of the plate to which the vacuum or pressure source is configured to be connected operatively.

5. The system of claim 1, wherein the vacuum or pressure source is a vacuum source configured to form a pressure sink at the output wells of the plate when the vacuum source is connected operatively to the output wells.

6. The system of claim 1, wherein the vacuum or pressure source is a pressure source configured to be connected operatively to input wells of the plate.

7. The system of claim 1, wherein the first fluid provides a continuous phase of the emulsion, wherein the wells of each unit include only one first input well configured to hold the first fluid, and wherein a pair of the channels of each unit extends to the droplet generator of such unit from the only one first input well.

8. The system of claim 1, wherein the plate includes a linear array of three or more emulsion production units.

9. The system of claim 1, wherein wells of the plate are spaced according to a well spacing of a standard microplate.

10. The system of claim 1, wherein the second fluid provides a dispersed phase of the emulsion, and wherein the second input well of each unit is configured to hold the second fluid and is disposed between the first input well and the output well of such unit.

11. The system of claim 10, wherein the first input well, the second input well, and the output well of each unit are arranged along a same line.

12. The system of claim 1, wherein each channel of each unit extends to the droplet generator of such unit from a bottom region of a well.

13. The system of claim 1, wherein the plate includes an upper member attached to a lower member, wherein the upper member forms side walls of the wells of each unit and also forms top and side walls of each channel of each unit, and wherein the lower member extends under each well and channel to form a bottom wall of such well and channel.

14. The system of claim 13, wherein the upper member is formed of an injection-molded polymer.

15. The system of claim 14, wherein each of the upper and lower members is formed by a respective, continuous piece of material.

16. The system of claim 1, wherein the plate includes an upper member attached to a lower member, wherein the upper member includes upper and lower surfaces, wherein the upper member defines through-holes corresponding to the wells of each unit and extending from the upper surface to the lower surface and also defines grooves corresponding to the channels of each unit and formed in the lower surface, and wherein the lower member is attached to the upper member at the lower surface to form a bottom wall below each through-hole and groove.

17. The system of claim 16, wherein the lower member is a sheet of material that is substantially thinner than the upper member.

18. The system of claim 1, wherein the first input well of a unit contains a nonaqueous continuous phase, wherein the second input well of such unit contains an aqueous phase configured for PCR amplification, and wherein the output well of such unit contains an emulsion including droplets of the aqueous phase disposed in the nonaqueous continuous phase.

19. The system of claim 1, wherein the plate includes an upper member attached to a lower member to form an array of emulsion production units each configured to produce a separate emulsion, and wherein the lower member has an upper surface that is flat and that abuts a lower surface of the upper member to form a bottom wall of openings formed in the lower surface and corresponding to the wells and the channels of each unit.

20. The system of claim 19, wherein each of the upper and lower members is formed by a respective, continuous piece of material.

21. The system of claim 1, wherein the at least one first input well of each emulsion production unit is not shared with other emulsion production units of the plate.

22. A method of forming an array of emulsions, comprising:

selecting a plate including a plurality of emulsion production units, each unit including

at least two input wells,

an output well, and

a droplet generator disposed along a flow path between the input wells and the output well;

disposing a first fluid in at least one first input well and a second fluid in a second input well of each unit; and

creating a pressure drop between the input wells and the output well of each unit such that the first fluid and the second fluid of each unit are driven from the input wells, through the droplet generator, and to the output well of such unit, for collection as an emulsion including droplets of the second fluid disposed in the first fluid.

23. The method of claim 22, wherein the step of selecting a plate includes a step of selecting a plate having the droplet generator of each unit formed by a site of channel intersection, and wherein the site of channel intersection is formed in part by one or more channels extending to the site from the first input well of the unit.

24. The method of claim 23, wherein the site of channel intersection is formed in part by a pair of channels extending to the site of channel intersection from a first input well of the unit.

25. The method of claim 22, wherein the step of selecting a plate includes a step of selecting a plate having only two input wells for each unit.

26. The method of claim 22, wherein the step of selecting a plate includes a step of selecting a plate that includes a linear array of three or more emulsion production units.

27. The method of claim 22, wherein the step of selecting a plate includes a step of selecting a plate having wells of the plate spaced according to a well spacing of a standard microplate.

28. The method of claim 22, wherein the step of disposing includes a step of disposing an aqueous phase in an input well of each unit with such input well being opposingly flanked by another input well and an output well of such unit.

29. The method of claim 22, wherein the step of disposing includes a step of disposing a liquid comprising a fluorocarbon oil in at least one input well of each unit.

30. The method of claim 22, wherein the step of creating a pressure drop drives the first fluid and the second fluid of each unit through a droplet generator structured at least substantially as a cross.

31. The method of claim 22, wherein the step of creating a pressure drop drives the first fluid of each unit into a pair of channels extending to the droplet generator from a single input well of such unit holding the first fluid.

32. The method of claim 22, wherein the step of creating a pressure drop includes a step of applying pressure to the input wells of each unit.

33. The method of claim 22, wherein the step of creating a pressure drop includes a step of applying a vacuum to the output well of each unit.

34. The method of claim 33, wherein the step of applying a vacuum causes production of separate emulsions in parallel by the units.

35. The method of claim 22, wherein the step of creating a pressure drop includes a step of engaging the plate with one or more gaskets to form a sealed relation between the one or more gaskets and at least one well of each unit.

36. The method of claim 22, wherein the step of disposing includes a step of disposing a second fluid that is an aqueous phase configured for PCR amplification.

37. The method of claim 22, wherein the plate is a first plate, wherein the step of creating a pressure drop is performed by an instrument, and wherein the step of creating a pressure drop is repeated using the instrument and a second plate including a plurality of emulsion production units.

38. The method of claim 22, wherein the step of creating a pressure drop is performed by an instrument including a manifold operatively connected to a vacuum or pressure source, and wherein the step of creating a pressure drop is performed without contact between the manifold and the first fluid, the second fluid, or the emulsion of any of the units.

39. The method of claim 22, wherein the step of disposing includes a step of disposing a first fluid including a surfactant and a second fluid including a surfactant.

40. The method of claim 22, wherein the step of creating a pressure drop includes a step of controlling air pressure in wells of the plate.

41. A method of forming an array of emulsions, comprising:

selecting a plate including a plurality of emulsion production units each including

only two input wells,

an output well, and

a droplet generator formed at an intersection of channels extending from the input wells to the output well;

disposing, in respective input wells of each unit, a continuous phase and an aqueous phase that is configured for PCR amplification; and

applying a vacuum in parallel to the output wells of the units such that the continuous phase and the aqueous phase of each unit are driven from the input wells, through the droplet generator, and to the output well, for collection as an emulsion including droplets of the aqueous phase disposed in the continuous phase.

42. The method of claim 41, wherein the step of selecting a plate includes a step of selecting a plate that includes a linear array of three or more emulsion production units.

43. The method of claim 41, wherein the step of disposing includes a step of disposing an aqueous phase in an input well of each unit with such input well being opposingly flanked by another input well and an output well of such unit.

44. The method of claim 41, wherein the step of applying a vacuum draws the continuous phase and the aqueous phase of each unit through a droplet generator structured at least substantially as a cross.

45. A method of forming an array of emulsions, comprising:

selecting a plate including a continuous upper member that is sealed underneath by a sheet of material having a flat upper surface, to form an array of three or more emulsion production units, each unit including

a pair of input wells,

an output well, and

a droplet generator structured at least substantially as a cross at an intersection of channels extending from the input wells to the output well;

disposing, in respective input wells of each unit, a continuous phase and an aqueous phase that is configured for PCR amplification; and

creating a pressure drop between the input wells and the output well of each unit such that the continuous phase and the aqueous phase of each unit are driven from the input wells, through the droplet generator, and to the output well, for collection as an emulsion including droplets of the aqueous phase disposed in the continuous phase.

46. The method of claim 45, wherein the step of creating a pressure drop includes a step of applying a vacuum to the output wells of the units.

47. The method of claim 45, wherein the step of creating a pressure drop includes a step of applying pressure to the inputs wells of the units.

48. The method of claim 45, wherein the step of creating a pressure drop causes production of separate emulsions in parallel by the units.