Device for generating droplets

• US 10,258,988 B2
• Filed: 11/30/2016
• Issued: 04/16/2019
• Est. Priority Date: 09/23/2008
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Bio-Rad Laboratories Incorporated
• Inventors: Ness, Kevin D., Hindson, Benjamin J., Colston, Jr., Billy W., Masquelier, Donald A.
• Primary Examiner(s): Hobbs, Michael L
• Application Number:
  US15/365,883
• Publication Number:
  US 20170144160A1
• Time in Patent Office: 867 Days
• 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

Devices and methods for generating droplets. An exemplary device comprises a substantially planar base portion including a bottom surface having a plurality of microfluidic channels formed therein as recessed regions of the bottom surface. The device also comprises a plurality of protrusions projecting from a top surface of the base portion and each formed integrally with the base portion. The device further comprises a sample well, a carrier well, and a droplet well. Each well has an upper portion created by one of the protrusions. A cover layer is attached to the bottom surface of the base portion and seals a bottom side of each microfluidic channel.

Claims

1. A device for generating droplets, comprising:

a substantially planar base portion including a bottom surface having a plurality of microfluidic channels formed therein as recessed regions of the bottom surface;

a plurality of protrusions projecting from a top surface of the base portion and each formed integrally with the base portion;

a sample well, a carrier well, and a droplet well, wherein each well has an upper portion created by one of the protrusions; and

a cover layer attached to the bottom surface of the base portion and sealing a bottom side of each microfluidic channel;

wherein the plurality of microfluidic channels includes a first channel, a second channel, and a third channel that meet one another in a droplet-generation region, and

wherein the sample well is configured to supply sample-containing fluid to the droplet-generation region via the first channel, the carrier well is configured to supply carrier fluid to the droplet-generation region via the second channel, and the droplet well is configured to receive sample-containing droplets in carrier fluid from the droplet-generation region via the third channel.

2. The device of claim 1, wherein the cover layer is thinner than the base portion.

3. The device of claim 1, wherein the base portion and the plurality of protrusions are provided by an injection-molded polymer body.

4. The device of claim 3, wherein each well defines a central axis and has a floor and a side wall region, wherein the side wall region surrounds the central axis and extends upwardly from the floor, and wherein the polymer body provides the side wall region of each well.

5. The device of claim 4, wherein the polymer body provides the floor of each well.

6. The device of claim 4, wherein the polymer body defines a plurality of openings each extending completely through the body in a direction orthogonal to the bottom surface of the base portion, and wherein each opening creates the side wall region of one of the wells.

7. The device of claim 6, wherein each opening narrows at a position intermediate a top side of the body and the bottom surface of the base portion.

8. The device of claim 1, where the plurality of microfluidic channels includes a fourth channel that meets the first channel, the second channel, and the third channel in the droplet-generation region.

9. The device of claim 8, wherein the carrier well is configured to supply carrier fluid to the droplet-generation region via the second channel and the fourth channel.

10. The device of claim 1, wherein the device comprises another well that is fluidically connected to the droplet well via the droplet-generation region.

11. A method of generating droplets, the method comprising:

selecting the device of claim 1; and

forming sample-containing droplets in the droplet-generation region.

12. A device for generating droplets, comprising:

an upper member injection molded as a single piece, the upper member defining a plane and a plurality of microfluidic channels arranged parallel to the plane;

a sample well, a carrier well, and a droplet well, each well defining a central axis and including a floor and a side wall region, the side wall region being formed by the upper member, the side wall region extending upwardly from the floor and surrounding the central axis; and

a lower member attached to the upper member and sealing a bottom side of each microfluidic channel;

wherein the plurality of microfluidic channels includes a first channel, a second channel, and a third channel that meet one another in a droplet-generation region, and wherein the sample well is configured to supply sample-containing fluid to the droplet-generation region via the first channel, the carrier well is configured to supply carrier fluid to the droplet-generation region via the second channel, and the droplet well is configured to receive sample-containing droplets in carrier fluid from the droplet-generation region via the third channel.

13. The device of claim 12, wherein the upper member includes a substantially planar base portion having a bottom surface at which the lower member is attached to the upper member, and wherein the lower member is thinner than the base portion.

14. The device of claim 12, wherein the upper member defines a plurality of openings each extending completely through the upper member, wherein the lower member covers a bottom end of each opening, and wherein each opening creates the side wall region of one of the wells.

15. The device of claim 14, wherein each opening narrows at a position intermediate a top side and a bottom surface of the upper member.

16. The device of claim 12, wherein the device comprises another well that is fluidically connected to the droplet well via the droplet-generation region.

17. The device of claim 12, wherein the device includes a plurality of sample wells, a plurality of carrier wells, a plurality of droplet wells, and a plurality of droplet-generation regions, and wherein each droplet-generation region is connected to one of the sample wells, one of the carrier wells, and one of the droplet wells.

18. A method of generating droplets, the method comprising:

selecting the device of claim 12; and

forming sample-containing droplets in the droplet-generation region.

19. A device for generating droplets, comprising:

a body injection molded as a single piece, the body including a base portion and a plurality of annular protrusions projecting from a top surface of the base portion, the base portion defining a plurality of microfluidic channels;

a sample well, a carrier well, and a droplet well, wherein each well has an upper portion created by one of the annular protrusions; and

a cover layer attached to the base portion and sealing a bottom side of each microfluidic channel;

wherein the plurality of microfluidic channels includes a first channel, a second channel, and a third channel that meet one another in a droplet-generation region, and

wherein the sample well is configured to supply sample-containing fluid to the droplet-generation region via the first channel, the carrier well is configured to supply carrier fluid to the droplet-generation region via the second channel, and the droplet well is configured to receive sample-containing droplets in carrier fluid from the droplet-generation region via the third channel.

20. The device of claim 19, wherein the cover layer is thinner than the base portion.

21. The device of claim 19, wherein each well defines a central axis and has a floor and a side wall region, wherein the side wall region surrounds the central axis and extends upwardly from the floor, and wherein the body provides the side wall region of each well.

22. The device of claim 19, where the plurality of microfluidic channels includes a fourth channel that meets the first channel, the second channel, and the third channel in the droplet-generation region.

23. The device of claim 19, wherein the carrier well is configured to supply carrier fluid to the droplet-generation region via the second channel and the fourth channel.

24. The device of claim 19, wherein the device includes a plurality of sample wells, a plurality of carrier wells, a plurality of droplet wells, and a plurality of droplet-generation regions, and wherein each droplet-generation region is connected to one of the sample wells, one of the carrier wells, and one of the droplet wells.

25. The device of claim 19, wherein the device comprises another well that is fluidically connected to the droplet well via the droplet-generation region.

26. A method of generating droplets, the method comprising:

selecting the device of claim 19; and

forming sample-containing droplets in the droplet-generation region.