METHOD AND SYSTEM FOR TRANSFERRING AND/OR CONCENTRATING A SAMPLE

• US 20100294665A1
• Filed: 07/11/2008
• Published: 11/25/2010
• Est. Priority Date: 07/12/2007
• Status: Abandoned Application

• Current Assignee: U S Genomics Incorporated
• Original Assignee: U S Genomics Incorporated
• Inventors: Zhou, Yi, Allen, Richard, Crane, Bryan, Burton, Randall E., Krogmeier, Jeffrey R., Sabourin, Nicaulas, Larson, Jonathan W., Papkov, Vyacheslav, Zhong, Qun
• Application Number:
  US12/668,579
• Publication Number:
  US 20100294665A1
• Time in Patent Office: Days
• US Class Current: 204/520
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 2001/4038 electric methods, e.g. electromigration, electrophoresis, ionisation
• CPC Class Codes: G01N 2001/4088 filtration

Abstract

A system for transferring and/or concentrating a sample is provided. The system includes a chamber with a membrane brave positioned within the chamber. An electrode assembly is configured to create an electric field across the membrane to move a charged sample through the chamber such that the sample collects and may concentrate on the membrane. The system may include a plurality of membranes. The system may also include a plurality of microchannels outwardly extending from the channel, where the membrane extends along the plurality of microchannels. Aspects of the invention are also directed to a pipette which may be used to transfer and concentrate a sample on a membrane. Certain embodiments are directed to methods and systems for concentrating a nucleic acid sample.

Claims

1. A system for concentrating a sample, the system comprising:

a first concentrator, the first concentrator comprising;

a first chamber;

a first semi-permeable membrane positioned within the first chamber, the first membrane having a first sample contacting surface;

an electrode assembly configured to selectively create an electric field across the first membrane to move a sample at least partially through the first chamber, from a starting position to the first sample contacting surface;

a second concentrator positioned downstream of the first concentrator, the second concentrator comprising;

a second chamber, wherein the second chamber is in selective fluid communication with the first chamber;

a second semi-permeable membrane positioned within the second chamber, the second membrane having a second sample contacting surface;

wherein the electrode assembly is also configured to selectively create an electric field across the second membrane to move a sample at least partially through the second chamber from an intermediate position to the second sample contacting surface; and

wherein the area of the first sample contacting surface is greater than the area of the second sample contacting surface.

2. The system of claim 1, wherein the ratio between the first sample contacting surface and the second sample contacting surface is at least approximately 2.

3. The system of claim 2, wherein the ratio between the first sample contacting surface and the second sample contacting surface is at least approximately 10.

4. The system of claim 3, wherein the ratio between the first sample contacting surface and the second sample contacting surface is at least approximately 20.

5. The system of claim 1, wherein the electrode assembly comprises:

a first electrode configured to selectively create an electric field across the first membrane; and

a second electrode configured to selectively create an electric field across the second membrane.

6. The system of claim 1, wherein the first and second membranes are polymer membranes.

7. The system of claim 1, further comprising a starting membrane in a reaction chamber positioned upstream of the first concentrator, wherein the starting membrane defines the sample starting position.

8. The system of claim 1, further comprising a plurality of substantially parallel microchannels outwardly extending from at least a portion of the first chamber, wherein the first membrane extends along the plurality of microchannels such that an electric field is selectively created through the plurality of microchannels.

9. The system of claim 8, wherein the plurality of substantially parallel microchannels includes a first set of substantially parallel microchannels and a second set of substantially parallel microchannels, wherein the first and second set are positioned on opposing sides of the first chamber.

10. The system of claim 1, further comprising a plurality of substantially parallel microchannels outwardly extending from at least a portion of the second chamber, wherein the second membrane extends along the plurality of microchannels such that an electric field is selectively created through the plurality of microchannels.

11. The system of claim 1, wherein the first concentrator is formed on a first micro-fluidic chip and the second concentrator is formed on a second micro-fluidic chip.

12. The system of claim 11, wherein the first concentrator includes a shallow depth region positioned between the first membrane and the electrode assembly such that the electric field is selectively created through the shallow depth region.

13. A method of transferring and concentrating a sample, the method comprising acts of:

providing the system for concentrating a sample as recited in claim 1;

creating an electric field with the electrode assembly across the first membrane such that a negatively charged sample moves from a starting position to the first sample contacting surface, whereby the concentration of the sample is greater on the first sample contacting surface than at the starting position.

14. The method of claim 13, further comprising the act of:

reversing the electric field to remove the negatively charged sample from the first sample contacting surface.

15. The method of claim 13, wherein the sample is a charged nucleic acid sample.

16. A system for concentrating a sample, the system comprising:

a chamber having an inlet port;

a plurality of substantially parallel microchannels outwardly extending from the channel;

a semi-permeable membrane extending within the chamber, the membrane having a first sample contacting surface, wherein the membrane extends along the plurality of microchannels;

an electrode assembly configured to selectively create an electric field through the plurality of microchannels and across the membrane to move a sample at least partially through the chamber, from a starting position to the first sample contacting surface.

17. The system of claim 16, wherein the membrane extends at least partially into the plurality of microchannels.

18. The system of claim 16, wherein the membrane is spaced apart from the plurality of microchannels.

19. The system of claim 18, further comprising a backside flow branch positioned between the membrane and the microchannels to provide fluid flow adjacent the membrane.

20. The system of claim 16, wherein the membrane is a polymer membrane.

21. The system of claim 16, wherein the chamber and the plurality of microchannels are pre-etched into a microfluidic chip.

22. The system of claim 16, wherein the plurality of microchannels are arranged to create a substantially uniform current density across the membrane when the electrode assembly creates an electric field.

23. The system of claim 16, further comprising a channel opposing the first sample contacting surface, wherein the electrode assembly is configured to selectively create an electric field into the channel opposing the first sample contacting surface to move a sample away from the first sample contacting surface.

24. A pipette, comprising:

a body having a passageway therethrough, the body having a first end and a second end;

a semi-permeable membrane positioned within the passageway, the semi-permeable membrane being spaced apart from the first end of the body;

an electrolytic buffer solution within the passageway; and

a positive electrode positioned at the second end of the body, wherein the positive electrode may selectively create an electric field through the passageway to the first end of the body when used in association with a negative electrode.

25. The pipette of claim 24, wherein the semi-permeable membrane is formed by a gel matrix plug.

26. The pipette of claim 24, wherein the semi-permeable membrane is formed by a regenerated cellulose membrane.

27. The pipette of claim 24, further comprising an organic solvent layer positioned within the passageway adjacent the semi-permeable membrane and proximate the first end of the body.

28. The pipette of claim 26, wherein the organic solvent layer is butanol.

29. The pipette of claim 25, wherein the gel matrix plug is formed with an acrylimide gel.

30. The pipette of claim 24, wherein the semi-permeable membrane is substantially resistant to the flow of a nucleic acid through the semi-permeable membrane.

31. The pipette of claim 24, further comprising a negative electrode positioned proximate the first end of the body, wherein the negative electrode and positive electrode create an electric field through the passageway.

32. The pipette of claim 24, wherein the positive electrode is positioned within the body.

33. A method of transferring and concentrating a sample, the method comprising acts of:

providing a pipette having a body with a passageway therethrough, a semi-permeable membrane positioned within the passageway, and an electrolytic buffer solution within the passageway;

placing a first end of the pipette in contact with a negatively charged sample contained within a reservoir; and

creating an electric field through the passageway of the pipette body such that a first portion of the negatively charged sample is drawn into a first end of the pipette body, whereby the concentration of the first portion of the sample is greater than the concentration of the sample in the reservoir.

34. The method of claim 33, further comprising the act of:

reversing the electric field to dispense the first portion of the negatively charged sample from the pipette.

35. The method of claim 33, further comprising the act of:

hydrodynamically dispensing the first portion of the negatively charged sample from the pipette.

36. The method of claim 33, wherein placing a first end of the pipette in contact with the negatively charged sample contained within the reservoir comprises placing the first end of the pipette in contact with a reservoir of a nucleic acid sample.

37. The method of claim 36, wherein the first portion of the sample includes nucleic acids and the first portion is drawn into a first end of the pipette body without substantially shearing the nucleic acids.

38. A system for transferring a sample, the system comprising:

a first chamber;

a first semi-permeable membrane positioned within the first chamber, the first membrane having a first sample contacting surface;

an assembly configured to move a sample at least partially through the first chamber, from a starting position to the first sample contacting surface;

a second chamber, wherein the second chamber is in selective fluid communication with the first chamber;

a second semi-permeable membrane positioned within the second chamber, the second membrane having a second sample contacting surface;

wherein the assembly is also configured to move a sample at least partially through the second chamber from an intermediate position to the second sample contacting surface, wherein the assembly includes an electrode assembly configured to selectively create an electric field across the second membrane to move a sample to the second sample contacting surface; and

wherein the area of the first sample contacting surface is greater than the area of the second sample contacting surface.

39. The system of claim 38, wherein the assembly includes an electrode assembly configured to selectively create an electric field across the first membrane to move a sample to the first sample contacting surface.

40. The system of claim 38, wherein the assembly hydrodynamically moves a sample to the first sample contacting surface.

41. The system of claim 38, further comprising:

a third chamber, wherein the third chamber is in selective fluid communication with the second chamber;

a third semi-permeable membrane positioned within the third chamber, the third membrane having a third sample contacting surface;

wherein the assembly is also configured to move a sample at least partially through the third chamber from an intermediate position to the third sample contacting surface, wherein the electrode assembly is configured to selectively create an electric field across the third membrane to move a sample to the third sample contacting surface.

42. The system of claim 41, wherein the area of the second sample contacting surface is greater than the area of the third sample contacting surface.

45. A method of performing a reaction with a sample, the method comprising acts of:

providing the system as recited in claim 39;

placing a sample within the first chamber;

creating an electric field with the electrode assembly across the first membrane such that a sample moves to the first sample contacting surface;

placing a reagent within the first chamber; and

performing a reaction with the sample and the reagent.

46. The method of claim 45, wherein performing a reaction with the sample and the reagent comprises creating an electric field with the electrode assembly across the first membrane such that the reagent moves to the first sample contacting surface to react with the sample.

47. The method of claim 45, wherein performing a reaction with the sample and the reagent comprises hydrodynamically moving the reagent to the first sample contacting surface to react with the sample.

48. The method of claim 45, further comprising:

flushing excess reagent out of the first chamber.

49. The method of claim 45, wherein the reaction is a chemical reaction.

50. The method of claim 45, wherein the reaction is a biological reaction.

51. The method of claim 45, wherein the reagent includes at least one of fluorescent intercalating dyes, bisPNA tags, restriction endonucleases, and DNA-binding reagents.

52. The method of claim 45, wherein the sample comprises nucleic acids.

43. A method of transferring and concentrating a sample, the method comprising acts of:

providing a chamber with a semi-permeable membrane positioned within the chamber, with the membrane having a sample contacting surface, and a channel opposing the sample contacting surface;

placing a negatively charged sample within the chamber;

creating an electric field across the membrane such that the negatively charged sample is drawn through the chamber and to the sample contacting surface of the membrane;

creating an electric field into the channel opposing the sample contacting surface to move the negatively charged sample away from the sample contacting surface and towards the channel; and

creating an electric field between the chamber and the channel to compress the sample in the chamber.

44. The method of claim 43, further comprising the acts of:

creating an electric field across the membrane such that the negatively charged sample moves back toward the sample contacting surface; and

creating an electric field between the chamber and the channel to compress the sample.