APPARATUS AND METHOD

• US 20110036994A1
• Filed: 09/04/2008
• Published: 02/17/2011
• Est. Priority Date: 09/04/2007
• Status: Active Grant

• Current Assignee: Base4 Innovation Ltd.
• Original Assignee: Base4 Innovation Ltd.
• Inventors: Frayling, Cameron
• Granted Patent: US 8,865,455 B2
• Time in Patent Office: Days
• US Class Current: 250/459.1
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 21/65 Raman scattering
• CPC Class Codes: G01N 21/658 enhancement Raman, e.g. surface plasmons
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44756 Apparatus specially adapted therefor
• CPC Class Codes: Y10S 977/704 Nucleic acids, e.g. DNA or RNA
• CPC Class Codes: Y10S 977/733 Nanodiaphragm
• CPC Class Codes: Y10S 977/781 Possessing nonosized surface openings that extend partially into or completely through the host material

Abstract

An apparatus for investigating a molecule comprising a channel provided in a substrate, a metallic moiety capable of plasmon resonance which is associated with the channel in a position suitable for the electromagnetic field produced by the metallic moiety to interact with a molecule passing therethrough, means to induce a molecule to pass through the channel, means to induce surface plasmon resonance in the metallic moiety; and means to detect interaction between the electromagnetic field produced by the metallic moiety and a molecule passing through the channel. Methods of investigating molecules are also provided.

Claims

1. An apparatus for investigating a molecule comprising:

at least one channel provided in a substrate, the channel being suitable for the passage of the molecule therethrough;

at least one metallic moiety capable of plasmon resonance, the at least one metallic moiety being associated with said channel in a position suitable for the electromagnetic field produced by the metallic moiety to interact with a molecule passing therethrough;

means to induce a molecule to pass through the channel;

means to induce surface plasmon resonance in the metallic moiety; and

means to detect interaction between the electromagnetic field produced by the metallic moiety and a molecule passing through the channel.

2. An apparatus according to claim 1 comprising two reservoirs separated from each other by the substrate, the first reservoir suitable to receive a sample comprising the molecule to be investigated, and a second reservoir suitable for the molecule to pass into once it has passed through the channel.

3. An apparatus according to claim 1 wherein the channel is an aperture.

4. An apparatus according to claim 1 wherein the aperture has a diameter of from approximately 0.5 nm to 100 nm.

5. An apparatus according to claim 1 wherein the aperture has a diameter of from approximately 1 nm to 50 nm.

6. An apparatus according to claim 1 wherein the aperture has a diameter of from approximately 1 nm to 30 nm.

7. An apparatus according to claim 1 having a diameter of from 1 nm to 5 nm.

8. An apparatus according to claim 1 wherein the aperture comprises an organic pore.

9. An apparatus according to claim 1 wherein the aperture is provided by a gel.

10. An apparatus according to claim 1 wherein the aperture is provided in a substrate such as a film.

11. An apparatus according to claim 1 wherein the aperture is formed from a nanoporous material selected from the group consisting of porous gold, a colloidal crystal, a colloidal cavity and a metal toroid.

12. An apparatus according to claim 1 comprising a plurality of apertures passing through the substrate.

13. An apparatus according to claim 12 wherein the apertures are arranged in an array.

14. An apparatus according to claim 1 wherein the metallic moiety comprises gold, silver, copper or aluminium.

15. An apparatus according to claim 1 wherein the metallic moiety is substantially spherical.

16. An apparatus according to claim 1 wherein the metallic moiety is substantially annular.

17. An apparatus according to claim 1 wherein the metallic moiety is a polygonal prism.

18. An apparatus according to claim 1 wherein the metallic moiety has a diameter of from 50 nm to 150 nm.

19. An apparatus according to claim 1 wherein the metallic moiety is positioned so as to be 50 nm or less from the molecule as it passes through the channel.

20. An apparatus according to claim 1 wherein the means to induce a molecule to pass through the channel comprises one or more of means to induce electrophoresis, a micro-fluidics device, or a laser trapping device.

21. An apparatus according to claim 1 comprising control means to control the passage of the molecule towards, into and/or through the aperture.

22. An apparatus according claim 21 wherein the control means comprise a porous matrix through which the molecule passes, the porous matrix lying immediately adjacent to the entrance to the channel.

23. An apparatus according to claim 1 wherein the means to detect interaction of the metallic moiety and the molecule comprise means to detect fluorescence or means to detect Raman scattering.

24. An apparatus according to claim 23 wherein the means to detect fluorescence comprise a photomultiplier or an avalanche photodiode.

25. An apparatus according to claim 23 wherein the means to detect Raman scattering comprise a Raman spectrometer.

26. A method for investigating a molecule comprising the steps of:

providing a fluid sample comprising a molecule to be investigated;

inducing said molecule to pass through a channel provided in a substrate, the channel having an associated metallic moiety which is induced to have plasmon resonance; and

detecting interaction between the electromagnetic field produced by the metallic moiety and the molecule as it passes through the channel.

27. The method according to claim 26 wherein the molecule is a biological molecule.

28. The method according to claim 27 wherein the molecule is a nucleic acid.

29. The method according to claim 26 wherein the molecule comprises a label which interacts with the metallic moiety.

30. The method according to claim 29 wherein the label is a fluorophore.

31. The method according to claim 30 wherein the label is one of Rose Bengal, C₅-maleimide, C₂-maleimide, Rhodamine Red, C₂-meleimide, and Rhodamine Green.

32. The method according to claim 31 wherein the label has an emission peak at slightly lower energy than the localised surface plasmon resonance peak of the metallic moiety.

33. A substrate suitable for use in an apparatus for investigating a molecule comprising:

at least one channel provided in the substrate, the channel suitable for the passage of a molecule therethrough; and

at least one metallic moiety capable of having have plasmon resonance induced, the at least one metallic moiety being associated with said channel in a position such that the electromagnetic field produced by the metallic moiety is able to interact with a molecule passing therethrough.

34. A method of sequencing a nucleic acid comprising the steps of:

providing a fluid sample comprising the nucleic acid to be sequenced;

inducing a single nucleic acid molecule to pass in a linear conformation through a pore in a substrate, the pore having a diameter of 0.5-100 nm and having an associated annular metallic moiety which is induced to have plasmon resonance; and

detecting interaction between the electromagnetic field produced by the annular metallic moiety and the single nucleic acid molecule as it passes through the channel.

35. A method according to claim 34, wherein the pore has a diameter of 1-50 nm.

36. A method according to claim 34, wherein all of one or more of the four bases in the nucleic acid is labeled with a fluorophore.