System for detection of spaced droplets

• US 9,132,394 B2
• Filed: 03/25/2011
• Issued: 09/15/2015
• 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: Makarewicz, Anthony J. Jr., Hiddessen, Amy L.
• Primary Examiner(s): Beisner, William H
• Assistant Examiner(s): HENKEL, DANIELLE B
• Application Number:
  US13/072,673
• Publication Number:
  US 20110311978A1
• Time in Patent Office: 1,635 Days
• Field of Search: 435/287.2, 435/6.12, 435/4, 435/91.2, 435/183, 506/27, 73/530.1, 73/540.1, 73/614.1, 738/632.1, 204/450, 204/554, 137/602, 137/892, 137/896
• US Class Current: 1/1
• CPC Class Codes: B01F 23/41 Emulsifying
• 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: B01L 2200/0689 Sealing
• 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 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2400/0478 pistons
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0622 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 3/563 Joints or fittings in general F16L; Separable fluid transfer means to transfer fluids between at least two containers, e.g. connectors
• 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: G01N 21/645 Specially adapted constructive features of fluorimeters

Abstract

System, including methods and apparatus, for detection of spaced droplets.

Claims

1. A detection system for droplet-based assays, comprising:

a channel network defining a flow path for droplets extending through a confluence region configured to increase an average distance between droplets, and to an examination region disposed downstream of the confluence region; and

a detector operatively connected to the examination region,

wherein the channel network includes a droplet inlet channel configured to carry droplets to the confluence region, wherein the droplet inlet channel forms a tapered region and a neck region extending from the tapered region to the confluence region, wherein the tapered region is sized such that droplets enter the neck region in single file, wherein the flow path has a smaller diameter in the neck region than in the examination region, and wherein at least one dilution inlet channel meets the droplet inlet channel at the confluence region and is configured to provide a dilution fluid that increases the average distance between droplets.

2. The detection system of claim 1, wherein the length of the neck region is between one and two times the average diameter of the droplets.

3. The detection system of claim 1, wherein the neck region has a substantially uniform diameter.

4. The detection system of claim 1, further comprising a droplet outlet channel that extends from the confluence region, and wherein the droplet outlet channel has about a same diameter as the neck region.

5. The detection system of claim 1, wherein the tapered region of the droplet inlet channel converges from a diameter of two or more average droplet diameters to a diameter of one or less than one average droplet diameters.

6. The detection system of claim 1, wherein the confluence region is formed at a junction between the droplet inlet channel, the at least one dilution inlet channel, and a droplet outlet channel.

7. The detection system of claim 6, wherein the at least one dilution inlet channel includes a pair of dilution inlet channels.

8. The detection system of claim 1, further comprising a light source configured to illuminate the examination region, wherein the detector is configured to detect light received from the examination region.

9. The detection system of claim 1, further comprising a tip having an open end configured to be placed into a container holding an emulsion such that the open end contacts the emulsion, wherein the flow path for droplets extends from the open end, through the confluence region, and to the examination region.

10. The detection system of claim 9, wherein the tip extends downward to the open end.

11. The detection system of claim 9, wherein the tip has a substantially uniform diameter.

12. The detection system of claim 1, wherein the diameter of the neck region is approximately equal to a diameter of the droplets.

13. A detection system for droplet-based assays, comprising:

a tip extending downward to an open end configured to be placed into a container holding an emulsion such that the open end contacts the emulsion;

a channel network defining a flow path for droplets, the flow path extending from the open end of the tip, through a confluence region configured to increase an average distance between droplets, and to an examination region disposed downstream of the confluence region; and

a detector operatively connected to the examination region,

wherein the channel network includes a droplet inlet channel configured to carry droplets to the confluence region, wherein the droplet inlet channel forms a tapered region and a neck region extending from the tapered region to the confluence region, and wherein the tapered region is sized such that droplets enter the neck region in single file.

14. The detection system of claim 13, wherein the length of the neck region is between one and two times the average diameter of the droplets.

15. The detection system of claim 13, wherein the neck region has a substantially uniform diameter.

16. The detection system of claim 13, further comprising a droplet outlet channel that extends from the confluence region, and wherein the droplet outlet channel has about a same diameter as the neck region.

17. The detection system of claim 13, wherein the tapered region of the droplet inlet channel converges from a diameter of two or more average droplet diameters to a diameter of one or less than one average droplet diameters.

18. The detection system of claim 13, wherein the confluence region is formed at a junction between the droplet inlet channel, a droplet outlet channel, and at least one dilution inlet channel configured to provide a dilution fluid.

19. The detection system of claim 18, wherein the at least one dilution inlet channel includes a pair of dilution inlet channels.

20. The detection system of claim 13, wherein the tip has a substantially uniform diameter.

21. The detection system of claim 13, wherein the diameter of the neck region is less than or approximately equal to a diameter of the droplets.

22. The detection system of claim 13, wherein the diameter of the neck region is approximately equal to the diameter of the droplets.

23. A method of detection for droplet-based assays using the detection system of claim 13, comprising:

driving droplets along the flow path from the tip, through the confluence region where a dilution fluid is introduced into the flow path, and through the examination region; and

detecting light from the examination region as droplets pass through.

24. The method of claim 23, further comprising a step of placing the open end of the tip into an emulsion, and a step of drawing droplets of the emulsion into the tip.

25. A detection system for droplet-based assays, comprising:

a channel network defining a flow path for droplets, the flow path extending through a confluence region configured to increase an average distance between droplets, and to an examination region disposed downstream of the confluence region; and

a detector operatively connected to the examination region,

wherein the channel network includes a droplet inlet channel configured to carry droplets to the confluence region, wherein the droplet inlet channel forms a tapered region and a neck region extending from the tapered region to the confluence region, wherein the tapered region is sized such that droplets enter the neck region in single file, and wherein the neck region has a diameter that is approximately equal to an average diameter of the droplets.

26. The detection system of claim 25, wherein the length of the neck region is between one and two times the average diameter of the droplets.

27. The detection system of claim 25, wherein the neck region has a substantially uniform diameter.

28. The detection system of claim 25, further comprising a droplet outlet channel that extends from the confluence region, and wherein the droplet outlet channel has about a same diameter as the neck region.

29. The detection system of claim 25, wherein the tapered region of the droplet inlet channel converges from a diameter of two or more average droplet diameters to a diameter of one or less than one average droplet diameters.

30. The detection system of claim 25, wherein the confluence region is formed at a junction between the droplet inlet channel, a droplet outlet channel, and at least one dilution inlet channel configured to provide a dilution fluid.

31. The detection system of claim 30, wherein the at least one dilution inlet channel includes a pair of dilution inlet channels.

32. The detection system of claim 25, further comprising a light source configured to illuminate the examination region, wherein the detector is configured to detect light received from the examination region.

33. The detection system of claim 25, further comprising a tip extending downward to an open end configured to be placed into a container holding an emulsion such that the open end contacts the emulsion, wherein the flow path for droplets extends from the open end, through the confluence region, and to the examination region.

34. The detection system of claim 25, wherein the flow path has a smaller diameter in the neck region than in the examination region.