Methods and devices for electronic and magnetic sensing of the contents of microfluidic flow channels

• US 8,440,093 B1
• Filed: 10/11/2006
• Issued: 05/14/2013
• Est. Priority Date: 10/26/2001
• Status: Expired due to Fees

• Current Assignee: Fluidigm Corporation
• Original Assignee: Fuidigm Corporation
• Inventors: Nassef, Hany, Facer, Geoff, Unger, Marc
• Primary Examiner(s): Tran, Binh X
• Application Number:
  US11/546,939
• Time in Patent Office: 2,407 Days
• Field of Search: 216/84, 216/86, 436/180, 324/71.1
• US Class Current: 216/84
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0655 pinch valves
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: C03C 25/68 by etching
• CPC Class Codes: G01N 15/1031 by measuring electrical or magnetic effects
• CPC Class Codes: G01N 15/12 by observing changes in resistance or impedance across apertures when traversed by individual particles, e.g. by using the Coulter principle
• CPC Class Codes: G01N 2015/1029 Particle size
• CPC Class Codes: G01N 27/00 Investigating or analysing materials by the use of electric, electrochemical, or magnetic means G01N3/00 – G01N25/00 take precedence; measurement or testing of electric or magnetic variables or of electric or magnetic properties of materials G01R
• CPC Class Codes: G01N 27/60 by investigating electrostatic variables , e.g. electrographic flaw testing G01N27/007 takes precedence
• CPC Class Codes: G01N 33/48707 by electrical means G01N33/49, G01N33/493 take precedence
• CPC Class Codes: G01N 33/48721 Investigating individual macromolecules, e.g. by translocation through nanopores Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

The presence of a detectable entity within a detection volume of a microfabricated elastomeric structure is sensed through a change in the electrical or magnetic environment of the detection volume. In embodiments utilizing electronic detection, an electric field is applied to the detection volume and a change in impedance, current, or combined impedance and current due to the presence of the detectable entity is measured. In embodiments utilizing magnetic detection, the magnetic properties of a magnetized detected entity alter the magnetic field of the detection volume. This changed magnetic field induces a current which can reveal the detectable entity. The change in resistance of a magnetoresistive element may also reveal the passage of a magnetized detectable entity.

Claims

1. A method of detecting entities of a selected entity type in a microfabricated structure, wherein the entity type is selected from bacterial cells, mammalian cells, and viral particles, the method comprising:

(a) providing a microfabricated structure including a flow channel, and a deflectable elastomeric membrane;

wherein the flow channel comprises a detection volume having selected physical dimensions with a selected width;

(b) applying an electric field across the detection volume;

(c) flowing a fluid containing a plurality of entities of the selected entity type through the detection volume at a selected flow rate;

(d) measuring a change in current and/or voltage of said applied electric field resulting from one of the entities in the fluid passing through and past the detection volume; and

(e) correlating said change(s) with the presence of an entity of the selected entity type within the flow channel,

wherein the method comprises selecting a microfabricated structure having the physical dimensions and width of the detection volume specified in step (a), in conjunction with a flow rate of the fluid through the detection volume specified in step (c), such that the combination of said physical dimensions and width of the flow channel and said flow rate result in only one entity of the selected entity type passing through the detection volume at a time.

2. The method of claim 1, wherein step (d) comprises measuring a change in voltage across the detection volume.

3. The method of claim 1, wherein step (d) comprises measuring a change in current across the detection volume.

4. The method of claim 1, wherein the detection volume is defined by electrodes separated from the flow channel by a dielectric material that permits an AC electrical field to be applied by the electrodes through the flow channel.

5. The method of claim 1, wherein the deflectable membrane is an elastomer with a Young'"'"'s modulus of 50 Pa to 10 MPa.

6. The method of claim 1, wherein the deflectable membrane is an elastomer with a Young'"'"'s modulus of about 1 MPa.

7. The method of claim 1, wherein the wherein the deflectable membrane comprises polydimethylsiloxane (PDMS).

8. The method of claim 1 wherein changes in the current and/or voltage are detected by electrodes positioned transversely on opposite sides of the detection volume.

9. The method of claim 8 wherein the same electrodes are used to apply the electric field and to detect the change in voltage and/or current across the detection volume.

10. The method of claim 8 wherein different electrodes are used to apply the electric field and to detect the change in voltage and/or current across the detection volume.

11. The method of claim 8 wherein the electrodes are in communication with an AC power supply such that an electric field having a constant frequency of oscillation is applied across the detection volume.

12. The method of claim 8 wherein the electrodes are in communication with an AC power supply such that said electric field varies in frequency of oscillation during measurement.

13. The method of claim 8 wherein the electrodes are in communication with a DC power supply such that a constant electric field is applied along the detection volume.

14. The method of claim 1 wherein changes in the current and/or voltage are detected by electrodes positioned at different points longitudinally along the flow channel.

15. The method of claim 14 wherein the same electrodes are used to apply the electric field and to detect the change in voltage and/or current along the detection volume.

16. The method of claim 14 wherein different electrodes a reused to apply the electric field and to detect the change in voltage and/or current across the detection volume.

17. The method of claim 14 wherein the electrodes are in communication with an AC power supply such that an electric field having a constant frequency of oscillation is applied along the detection volume.

18. The method of claim 14 wherein the electrodes are in communication with an AC power supply such that said electric field varies in frequency of oscillation during measurement.

19. The method of claim 14 wherein the electrodes are in communication with a DC power supply such that a constant electric field is applied along the detection volume.

20. A method of detecting a magnetized entity of a selected entity type in a microfabricated structure, wherein the entity type is selected from bacterial cells, mammalian cells, viral particles, microparticles, and nanoparticles, the method comprising:

(a) providing the microfabricated structure including a flow channel and a deflectable membrane;

wherein within the flow channel is a detection volume configured to receive one entity of the selected entity type or an ensemble thereof at a time;

wherein the microfabricated structure further comprises a conductive coil structure proximate to the detection volume;

(b) measuring a current and/or a voltage induced in the coil structure by passage of an entity of the selected entity type or an ensemble thereof through the detection volume; and

(c) correlating the current and/or voltage induced in the coil structure with the presence of an entity of the selected entity type within the flow channel.

21. The method of claim 20 wherein the conductive coil structure is formed in elastomeric material adjacent to the flow channel.

22. The method of claim 20 wherein the conductive coil structure is coaxial with the flow channel.

23. A method of detecting a magnetized entity of a selected entity type in a microfabricated structure, wherein the entity type is selected from bacterial cells, mammalian cells, viral particles, microparticles, and nanoparticles, the method comprising:

(a) providing the microfabricated structure including a flow channel and a deflectable membrane;

wherein within the flow channel is a detection volume configured to receive one entity of the selected entity type or an ensemble thereof at a time;

wherein the microfabricated structure further comprises a magnetoresistive element proximate to the detection volume;

(b) measuring a changed resistance across the magnetoresistive element caused by passage of an entity of the selected entity type or an ensemble thereof through the detection volume; and

(c) correlating said changed resistance with the presence of an entity of the selected entity type within the flow channel.

24. A method of detecting isolated macromolecules in a microfabricated structure, the method comprising:

(a) providing a microfabricated structure including a flow channel and a deflectable membrane;

wherein the flow channel comprises a detection volume having selected physical dimensions with a selected width;

(b) applying an electric field to the detection volume;

(c) flowing a fluid containing said isolated macromolecules through the detection volume at a selected flow rate;

(d) measuring a change in current and/or voltage of said applied electric field resulting from a macromolecule or ensemble thereof passing through and past the detection volume; and

(e) correlating said change(s) with the presence of a macromolecule within the flow channel,

wherein the method comprises selecting a microfabricated structure having the physical dimensions and width of the detection volume specified in step (a), in conjunction with a flow rate of the fluid through the detection volume specified in step (c), such that the combination of said physical dimensions and width of the flow channel and said flow rate result in only one of said macromolecules or an ensemble thereof passing through the detection volume at a time.

25. The method of claim 24 wherein said macromolecule is a nucleic acid or a protein.

26. The method of claim 24, wherein step c) comprises measuring a change in voltage across the detection volume, and step d) comprises correlating the change in voltage with the presence of a macromolecule within the flow channel.

27. The method of claim 24, wherein step c) comprises measuring a change in current across the detection volume, and step d) comprises correlating the change in current with the presence of a macromolecule within the flow channel.

28. The method of claim 24, wherein the detection volume is defined by electrodes separated from the flow channel by a dielectric material that permits an AC electrical field to be applied by the electrodes through the flow channel.

29. The method of claim 24 wherein changes in the current and/or voltage are detected by electrodes positioned transversely on opposite sides of the detection volume.

30. The method of claim 29 wherein the same electrodes are used to apply the electric field and to detect the change in voltage and/or current across the detection volume.

31. The method of claim 29 wherein different electrodes are used to apply the electric field and to detect the change in voltage and/or current across the detection volume.

32. The method of claim 29 wherein the electrodes are in communication with an AC power supply such that said electric field has a constant frequency of oscillation.

33. The method of claim 29 wherein the electrodes are in communication with an AC power supply such that said electric field varies in frequency of oscillation during measurement.

34. The method of claim 29 wherein the electrodes are in communication with a DC power supply such that a constant electric field is applied along the detection volume.

35. The method of claim 24 wherein changes in the current and/or voltage are detected by electrodes positioned at different points longitudinally along the flow channel.

36. The method of claim 35 wherein the same electrodes are used to apply the electric field and to detect the change in voltage and/or current across the detection volume.

37. The method of claim 35 wherein different electrodes are used to apply the electric field and to detect the change in voltage and/or current across the detection volume.

38. The method of claim 35 wherein the electrodes are in communication with an AC power supply such that said electric field has a constant frequency of oscillation.

39. The method of claim 35 wherein the electrodes are in communication with an AC power supply such that said electric field varies in frequency of oscillation during measurement.

40. The method of claim 35 wherein the electrodes are in communication with a DC power supply such that a constant electric field is applied along the detection volume.