System for forming an array of emulsions

• US 9,126,160 B2
• Filed: 12/08/2010
• Issued: 09/08/2015
• 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: Bio-Rad Laboratories Incorporated
• Inventors: Ness, Kevin D., Hindson, Benjamin J., Colston, Billy W. Jr., Masquelier, Donald A.
• Primary Examiner(s): HOBBS, MICHAEL L
• Application Number:
  US12/963,523
• Publication Number:
  US 20110086780A1
• Time in Patent Office: 1,735 Days
• Field of Search: 435/287.2, 506/23, 506/40
• US Class Current: 1/1
• 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 35/165 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

A system, including method and apparatus, for forming an array of emulsions. The system may include a plate providing an array of emulsion production units each configured to produce a separate emulsion and each including a set of wells interconnected by channels that intersect to form a site of droplet generation. Each set of wells, in turn, may include (1) at least one first input well to receive a continuous phase, (2) a second input well to receive a dispersed phase, and (3) an output well configured to receive from the site of droplet generation an emulsion of droplets of the dispersed phase disposed in the continuous phase.

Claims

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

a plate providing an array of emulsion production units each configured to produce a separate emulsion and each including a set of wells interconnected by a set of channels forming a channel junction, each channel being bounded circumferentially, each set of wells including

at least one first input well to receive a continuous phase,

a second input well to receive a dispersed phase, and

an output well;

wherein the set of channels includes at least two input channels extending separately from the input wells to the channel junction, at which droplets of the dispersed phase are generated in the continuous phase, and an output channel extending from the channel junction to the output well, in which an emulsion is collected.

2. The system of claim 1, wherein the set of wells of each unit includes only one first input well, and wherein a pair of the channels of the set of channels of each unit extend separately from one another to the channel junction of such unit from the only one first input well.

3. The system of claim 1, wherein only one of the channels of each unit extends from the second input well to the channel junction of such unit, and wherein only one of the channels of each unit extends from the channel junction to the output well of such unit.

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

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

6. The system of claim 1, wherein the second input well of each unit is disposed between the first input well and the output well of such unit.

7. The system of claim 6, wherein the first input well, the second input well, and the output well of each unit are arranged along a same line.

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

9. 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 the channels of each unit, and wherein the lower member extends under each well and channel of each unit to form a bottom wall of such well and channel.

10. The system of claim 9, wherein the upper member is formed of an injection-molded polymer.

11. The system of claim 9, wherein each of the upper and lower members is formed by a respective, continuous piece of material.

12. 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 under each through-hole and groove.

13. The system of claim 12, wherein the lower member is a sheet of material that is substantially thinner than the upper member.

14. 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.

15. 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.

16. The system of claim 1, further comprising an instrument configured to create a pressure drop between the input wells and the output well of each unit such that the continuous phase and the dispersed phase of each unit are driven from the input wells, through the channel junction, and to the output well of such unit, for collection as an emulsion including droplets of the dispersed phase disposed in the continuous phase.

17. The system of claim 16, wherein the instrument includes a vacuum source.

18. The system of claim 16, wherein the instrument includes a pressure source.

19. The system of claim 16, wherein the instrument is configured to operate the emulsion production units without contacting liquid contents of any wells of the units.

20. A system for forming an array of emulsions in parallel, comprising:

a plate having an upper member attached to a lower member to form an array of emulsion production units each configured to produce a separate emulsion, each unit including a set of wells interconnected by a set of channels forming a channel junction, each channel being bounded circumferentially, each set of wells including

at least one first input well to receive a continuous phase,

a second input well to receive a dispersed phase, and

an output well;

wherein the set of channels includes at least two input channels extending separately from the input wells to the channel junction, at which droplets of the dispersed phase are generated in the continuous phase, and an output channel extending from the channel junction to the output well, in which an emulsion is collected, 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.

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