Miniature support for thin films containing single channels or nanopores and methods for using the same

• US 6,746,594 B2
• Filed: 06/08/2001
• Issued: 06/08/2004
• Est. Priority Date: 11/06/1998
• Status: Expired due to Term

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation), Regents of the University of California (University of California)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation), Regents of the University of California (University of California)
• Inventors: Deamer, David W., Akeson, Mark A., Branton, Daniel
• Primary Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US09/877,745
• Publication Number:
  US 20020056651A1
• Time in Patent Office: 1,096 Days
• Field of Search: 204/450, 204/451, 204/518, 204/520, 204/409, 204/600, 204/601, 204/627, 204/630, 204/418, 204/789, 204/403.01, 204/403.06, 204/403.07, 204/403.08, 204/416, 205/775, 205/792, 205/793, 205/777.5, 422/50, 422/68.1, 422/82.01
• US Class Current: 205/777.5
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2565/631 being a biochannel or pore
• 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: Y10S 977/781 Possessing nonosized surface openings that extend partially into or completely through the host material
• CPC Class Codes: Y10S 977/788 Of specified organic or carbon-based composition
• CPC Class Codes: Y10S 977/932 for electronic or optoelectronic application

Abstract

Single-channel thin film devices and methods for using the same are provided. The subject devices comprise cis and trans chambers connected by an electrical communication means. At the cis end of the electrical communication means is a horizontal conical aperture sealed with a thin film that includes a single nanopore or channel. The devices further include a means for applying an electric field between the cis and trans chambers. The subject devices find use in applications in which the ionic current through a nanopore or channel is monitored, where such applications include the characterization of naturally occurring ion channels, the characterization of polymeric compounds, and the like.

Claims

1. A single-channel thin film device comprising:

a cis chamber;

a trans chamber;

an electrical communication means for holding a conductor of electrical current connecting said cis and trans chambers and having a cis terminus and a trans terminus; and

a single-channel horizontal sealed aperture at said cis terminus.

2. The device according to claim 1, wherein said single-channel horizontal sealed aperture comprises:

a conical aperture; and

a thin film sealing said aperture, wherein said thin film comprises a single nanopore.

3. The device according to claim 1, wherein said device further comprises a means for applying an electric field between said cis and trans chambers.

4. The device according to claim 2 wherein said nanopore is selected from the group consisting of a naturally occurring proteinaceous channel and a synthetic pore.

5. The device according to claim 1, wherein said electrical communication means is a U-shaped tube connecting said cis chamber with said trans chamber.

6. In a method in which the current through a nanopore of a single-channel horizontal sealed aperture is monitored, the improvement comprising:

monitoring current through a nanopore using the single-channel thin film device according to claim 1.

7. The method according to claim 6, wherein said method further includes characterizing a naturally occurring ion channel.

8. The method according to claim 6, wherein said method further includes characterizing a polymeric compound.

9. The method according to claim 8, wherein said method further includes sequencing a nucleic acid.

10. A single-channel thin film device comprising:

a cis chamber;

a trans chamber;

a U-shaped tube connecting said cis chamber with said trans chamber and having a cis terminus and a trans terminus;

a conical aperture at said cis terminus, wherein said aperture is sealed with a thin film comprising a single nanopore; and

a means for applying an electric field between said cis and trans chambers.

11. The device according to claim 10, wherein said conical aperture has an inner diameter ranging from about 1 to 50 μ

m.

12. The device according to claim 10, wherein said nanopore has an inner diameter ranging from about 1 to 10 nm.

13. The device according to claim 12, wherein said nanopore is selected from the group consisting of a naturally occurring protein channel and a synthetic pore.

14. The device according to claim 13, wherein said naturally occurring protein channel is a heptameric channel of α

-hemolysin.

15. The device according to claim 14, wherein said channel is a rectifying channel.

16. A method of monitoring ionic current through a nanopore, said method comprising:

applying an electrical field between the cis and trans chambers of a device according to claim 10, and

measuring the ionic current through said nanopore.

17. A single-channel thin film device comprising:

a cis chamber;

a trans chamber;

a U-shaped tube connecting said cis chamber with said trans chamber and having a cis terminus and a trans terminus;

a conical aperture at said cis terminus, wherein said conical aperture has an inner diameter ranging from about 1 to 50 μ

m and is sealed with a lipid bilayer comprising a single rectifying heptameric channel of α

-hemolysin; and

a means for applying an electric field between said cis and trans chambers.

18. The device according to claim 17, wherein said conical aperture is fabricated from Teflon.

19. The device according to claim 17, wherein said means for applying an electric field comprises a cis electrode and a trans electrode.