Method and apparatus for the discretization and manipulation of sample volumes

• US 9,180,453 B2
• Filed: 07/27/2009
• Issued: 11/10/2015
• 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:
  US12/510,110
• Publication Number:
  US 20100041046A1
• Time in Patent Office: 2,297 Days
• US Class Current: 1/1
• 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. A method comprising:

providing a fluidic lattice comprising,

a flow channel having a flow axis, and

a plurality of fluidic harbors in fluidic communication with the flow channel and offset from the flow axis;

flowing a primer continuous liquid through the fluidic lattice and into the flow channel and into the fluidic harbors;

flowing a first continuous liquid through the fluidic lattice to provide the first continuous liquid within the plurality of fluidic harbors, wherein the primer continuous liquid is immiscible with the first continuous liquid such that flowing the first continuous liquid through the fluidic lattice displaces at least a portion of the primer continuous liquid from the flow channel and from the plurality of fluidic harbors; and

flowing a second continuous liquid through the fluidic lattice, the second continuous liquid immiscible with the first continuous liquid such that the second continuous liquid displaces the first continuous liquid from the flow channel while allowing the first continuous liquid to remain in the plurality of fluidic harbors.

2. The method of claim 1 wherein the first continuous liquid comprises an aqueous solution.

3. The method of claim 1 wherein the second continuous liquid is flowed at a fast rate to displace some of the first continuous liquid from at least one of the plurality of fluidic harbors.

4. The method of claim 1 wherein the first continuous liquid is flowed at a fast rate to displace less than all of the primer continuous liquid from at least one of the plurality of fluidic harbors.

5. The method of claim 1 wherein the first continuous liquid comprises an analyte, the method further comprising performing analysis of the first continuous liquid within at least one of the plurality of fluidic harbors.

6. The method of claim 5 wherein the analyte comprises a biological material.

7. The method of claim 6 wherein the biological material is selected from a cell, a virus, a prion, a nucleic acid, a protein, a genetic material, an expressed product of a genetic material, a crystallizing molecule, or a particle.

8. The method of claim 6 wherein performing the analysis comprises performing biochemical analysis.

9. The method of claim 5 wherein the first continuous liquid comprises a plurality of strands of analyte nucleic acids and the plurality of fluidic harbors is configured such that either one or more analyte nucleic acid strands or no analyte nucleic acid strands are contained within each of the plurality of fluidic harbors, the method further comprising performing digital polymerase chain reaction (PCR) within the plurality of fluidic harbors.

10. The method of claim 5 wherein the analyte undergoes crystallization in the fluidic harbor.

11. The method of claim 5 wherein the analyte contains at least one rare cell selected from cells expressing a malignant phenotype, fetal cells, circulating endothelial cells, tumor cells, cells infected with a virus, cells transfected with a gene of interest, aberrant subtypes of T-cells present in the peripheral blood of subjects afflicted with autoimmune or autoreactive disorders, and aberrant subtypes of B-cells present in the peripheral blood of subjects afflicted with autoimmune or autoreactive disorders.

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

13. The method of claim 12 wherein a cross-section of at least one of the fluidic harbors is T-shaped, L-shaped, triangular, rectangular, circular, elliptical, polygonal, or square.

14. The method of claim 1 wherein at least one of the plurality of fluidic harbors is displaced in a vertical direction above the flow axis, in order to retain a buoyant species.

15. The method of claim 1 wherein at least one of the plurality of fluidic harbors is displaced in a vertical direction below the flow axis, in order to retain a dense species.

16. The method of claim 1 further comprising modifying a surface of at least one of the fluidic harbors to affect its interaction with the first continuous liquid and/or the second continuous liquid.

17. The method of claim 1 further comprising delivering a material or an energy to the at least one of the plurality of fluidic harbors through an auxiliary channel.

18. The method of claim 1 wherein the plurality of fluidic harbors are arranged in parallel, in series, or in both parallel and in series along the flow channel.

19. The method of claim 1 further comprising causing a contents of one of the plurality of fluidic harbors to mix with a chemical.

20. The method of claim 19 wherein the mixing results from breaking down a wall between the one of the plurality of fluidic harbors, and a chamber containing the chemical.

21. The method of claim 20 wherein the chamber is located out of a plane defined by the flow channel.

22. The method of claim 1 further comprising removing a contents including the first continuous liquid, from at least one of the plurality of fluidic harbors.

23. The method of claim 22 wherein the contents are removed through an access channel.

24. The method of claim 22 wherein the contents are removed by flowing another continuous liquid through the fluidic lattice, the another continuous liquid immiscible with the contents.

25. The method of claim 24 wherein a viscosity of the another continuous liquid is different from a viscosity of the contents.

26. The method of claim 24 wherein the another continuous liquid includes a surfactant.

27. The method of claim 22 wherein the contents are removed by an application of force to the contents of the at least one of the fluidic harbors.

28. The method of claim 27 wherein the force is selected from at least one of thermal energy, vacuum, pneumatic pressure, acoustic pressure, ultrasonic pulses, radiation pressure, electromagnetic field, electrowetting, photo-induced surface wetting, electrocapillarity, surface or interfacial tension, thermal-gradient-derived forces, electrostatic interaction, optical forces, or magnetic forces.

29. The method of claim 22 wherein the contents are removed utilizing a valve.

30. The method of claim 22 wherein the contents are removed by piercing the fluidic lattice with a needle.

31. The method of claim 22 further comprising further manipulating the contents removed from the one of the plurality of fluidic harbors.

32. The method of claim 1 wherein a continuous liquid comprises an undisrupted volume that is equal to or greater than the volume of two fluidic harbors.

33. The method of claim 1 wherein a continuous liquid comprises an undisrupted volume that is equal to or greater than the volume of five fluidic harbors.

34. The method of claim 1 wherein a continuous liquid comprises an undisrupted volume that is equal to or greater than the volume of 20 fluidic harbors.

35. The method of claim 1 wherein flowing the first continuous liquid through the fluidic lattice displaces a portion of the primer continuous liquid from a fluidic harbor such that a fluidic packet of the first continuous liquid is surrounded at least in part by a remaining portion of the primer continuous liquid in the fluidic harbor and the remaining portion of the primer continuous liquid contacts at least a portion of the surface of the fluidic harbor.

36. The method of claim 1 wherein at least one fluidic harbor comprises a drain channel in fluidic communication with the at least one fluidic harbor and with the flow channel.

37. A method of performing isothermal digital polymerase chain reaction, the method comprising:

providing a fluidic lattice comprising,

a flow channel having a flow axis, and

a plurality of fluidic harbors in fluidic communication with the flow channel and offset from the flow axis;

flowing a primer continuous liquid through the fluidic lattice and into the flow channel and into the fluidic harbors;

flowing a first continuous liquid through the fluidic lattice, wherein the primer continuous liquid is immiscible with the first continuous liquid such that flowing the first continuous liquid through the fluidic lattice displaces at least a portion of the primer continuous liquid from the flow channel and from the plurality of fluidic harbors, and wherein the first continuous liquid contains a plurality of strands of analyte nucleic acids such that either one or more analyte nucleic acid strands or no analyte nucleic acid strands are provided within each of the plurality of fluidic harbors;

flowing a second continuous liquid through the fluidic lattice, the second continuous liquid immiscible with the first continuous liquid such that the second continuous liquid displaces the first continuous liquid from the flow channel while allowing the first continuous liquid to remain in the plurality of fluidic harbors;

heating the first continuous liquid within the plurality of the fluidic harbors; and

detecting a concentration of the analyte nucleic acid strands within the plurality of the fluidic harbors.

38. A method of performing digital polymerase chain reaction (PCR), the method comprising:

providing a fluidic lattice comprising,

a flow channel having a flow axis, and

a plurality of fluidic harbors in fluidic communication with the flow channel and offset from the flow axis;

flowing a primer continuous liquid through the fluidic lattice and into the flow channel and into the fluidic harbors;

flowing a first continuous liquid through the fluidic lattice, wherein the primer continuous liquid is immiscible with the first continuous liquid such that flowing the first continuous liquid through the fluidic lattice displaces at least a portion of the primer continuous liquid from the flow channel and from the plurality of fluidic harbors, and wherein the first continuous liquid contains a plurality of strands of analyte nucleic acids such that either one or more analyte nucleic acid strands or no analyte nucleic acid strands are provided within each of the plurality of fluidic harbors;

flowing a second continuous liquid through the fluidic lattice, the second continuous liquid immiscible with the first continuous liquid such that the second continuous liquid displaces the first continuous liquid from the flow channel while allowing the first continuous liquid to remain in the plurality of fluidic harbors;

performing a plurality of heating cycles of the first continuous liquid within the plurality of the fluidic harbors; and

detecting a concentration of the analyte nucleic acid strands within the plurality of the fluidic harbors.