Method and apparatus for the discretization and manipulation of sample volumes

• US 10,421,070 B2
• Filed: 09/29/2015
• Issued: 09/24/2019
• Est. Priority Date: 08/15/2008
• Status: Active Grant

• Current Assignee: University of Washington
• Original Assignee: University of Washington
• Inventors: Chiu, Daniel T., Cohen, Dawn E., Jeffries, Gavin D. M.
• Primary Examiner(s): Forman, Betty J
• Application Number:
  US14/869,871
• Publication Number:
  US 20160096172A1
• Time in Patent Office: 1,456 Days
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/165 Specific details about hydrophobic, oleophobic surfaces
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1861 using radiation
• CPC Class Codes: B01L 2400/0439 ultrasonic vibrations, vibrating piezo elements
• CPC Class Codes: B01L 2400/0633 with moving parts
• CPC Class Codes: B01L 3/502 with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• 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: G01N 2035/00356 Holding samples at elevated temperature (incubation)
• CPC Class Codes: G01N 2035/1032 Dilution or aliquotting
• CPC Class Codes: G01N 2035/1034 Transferring microquantities of liquid
• 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: G01N 35/10 Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices G01N35/0099 takes precedence

Abstract

Embodiments of the present invention relate to methods and apparatuses for the discretization and manipulation of sample volumes that is simple, robust, and versatile. It is a fluidic device that partitions a sample by exploiting the interplay between fluidic forces, interfacial tension, channel geometry, and the final stability of the formed droplet and/or discretized volume. These compartmentalized volumes allow for isolation of samples and partitioning into a localized array that can subsequently be manipulated and analyzed. The isolation of the discretized volumes along with the device'"'"'s inherent portability render our invention versatile for use in many areas, including but not limited to PCR, digital PCR, biological assays for diagnostics and prognostics, cancer diagnosis and prognosis, high throughput screening, single molecule and single cell reactions or assays, the study crystallization and other statistical processes, protein crystallization, drug screening, environmental testing, and the coupling to a wide range of analytical detection techniques for biomedical assays and measurements. The minimal fluid interconnects and simple flow geometry makes the device easy to use and implement, economical to fabricate and operate, and robust in its operations.

Claims

1. An apparatus comprising:

a flow channel having a flow axis, the flow channel being in selective fluid communication with a source of a continuous fluid comprising an aqueous solution;

a first fluidic harbor offset from the flow axis and fluidically coupled to the flow channel at a first location of the flow channel and at a second location of the flow channel;

a second fluidic harbor offset from the flow axis and fluidicially coupled to the flow channel at a first plurality of locations upstream of the first location along the flow channel;

a third fluidic harbor offset from the flow axis and fluidically coupled to the flow channel at a second plurality of locations downstream of the second location along the flow channel; and

a primer solution immiscible with the aqueous solution, the primer solution disposed within at least two of the fluidic harbors;

wherein the first location is upstream of the second location along the flow channel.

2. The apparatus of claim 1 wherein the flow channel is in selective fluid communication with a source of a primer fluid.

3. The apparatus of claim 2 wherein the primer fluid contains a surfactant.

4. The apparatus of claim 2 wherein a surface of at least one of the fluidic harbors is modified to affect its interaction with the continuous fluid and/or the primer fluid.

5. The apparatus of claim 1 wherein the continuous fluid is an aqueous solution comprising an analyte.

6. The apparatus of claim 5 wherein the analyte is a nucleic acid.

7. The apparatus of claim 5 wherein the analyte is a biological cell.

8. The apparatus of claim 1 wherein the flow channel contains a primer fluid and the primer fluid contains a surfactant.

9. The apparatus of claim 1 wherein a shape of at least one of the plurality of fluidic harbors is other than spherical.

10. The apparatus of claim 1 wherein the plurality of fluidic harbors are arranged in series along the flow channel.

11. The apparatus of claim 1 further comprising a second channel in fluid communication with at least two fluidic harbors of the plurality of fluidic harbors.

12. The apparatus of claim 1 further comprising a second channel whose dimension is smaller than the flow channel.

13. The apparatus of claim 1 wherein a surface of at least one of the fluidic harbors is hydrophobic.

14. The apparatus of claim 1 wherein at least one of the fluidic harbors is oleophilic.

15. The apparatus of claim 1 wherein at least one of the fluidic harbors is located out of a plane defined by the flow channel.

16. The apparatus of claim 1 wherein the fluidic harbor comprises a nucleic acid.

17. An apparatus comprising:

a flow channel having a flow axis, the flow channel being in selective fluid communication with a source of a first continuous liquid and in selective fluid communication with a source of a second continuous liquid immiscible with the first liquid;

a first fluidic harbor offset from the flow axis and fluidically coupled to the flow channel at a first location of the flow channel and at a second location of the flow channel, wherein the first location is upstream of the second location along the flow channel;

a second fluidic harbor offset from the flow axis and fluidically coupled to the flow channel at a first plurality of locations upstream of the first location along the flow channel;

a third fluidic harbor offset from the flow axis and fluidically coupled to the flow channel at a second plurality of locations downstream of the second location along the flow channel; and

a primer fluid contained within at least one of the plurality of fluidic harbors, wherein the primer fluid is immiscible with the first continuous liquid.

18. The apparatus of claim 17 wherein the flow channel is in selective fluid communication with a source of a primer fluid.

19. The apparatus of claim 18 wherein the primer fluid contains a surfactant.

20. The apparatus of claim 17 wherein the flow channel contains a primer fluid.

21. The apparatus of claim 20 wherein the primer fluid contains a surfactant.

22. The apparatus of claim 17 wherein the first continuous liquid is an aqueous solution.

23. The apparatus of claim 22 wherein the aqueous solution contains an analyte.

24. The apparatus of claim 23 wherein the analyte is a nucleic acid.

25. The apparatus of claim 23 wherein the analyte is a biological cell.

26. The apparatus of claim 17 wherein the second continuous liquid contains a surfactant.

27. The apparatus of claim 17 wherein a shape of at least one of the plurality of fluidic harbors is other than spherical.

28. The apparatus of claim 27 wherein a cross-section of at least one of the fluidic harbors is T-shaped, L-shaped, triangular, rectangular, or square.

29. The apparatus of claim 17 wherein a surface of at least one of the fluidic harbors is modified to affect its interaction with the first liquid and/or the second liquid.

30. The apparatus of claim 17 wherein the plurality of fluidic harbors are arranged in series along the flow channel.

31. The apparatus of claim 17 further comprising a second channel in fluid communication with at least two fluidic harbors of the plurality of fluidic harbors.

32. The apparatus of claim 17 wherein a surface of at least one of the fluidic harbors is hydrophobic.

33. The apparatus of claim 17 wherein at least one of the fluidic harbors is oleophilic.

34. The apparatus of claim 17 wherein at least one of the fluidic harbors is located out of a plane defined by the flow channel.

35. The apparatus of claim 34 wherein at least one fluidic harbor has a height taller than the height of the flow channel.

36. The apparatus of claim 17 wherein the fluidic harbor comprises a nucleic acid.

37. The apparatus of claim 17 wherein the primer fluid and the second continuous liquid comprise the same fluid.