NANOSCALE APERTURES HAVING ISLANDS OF FUNCTIONALITY

• US 20170253480A1
• Filed: 03/24/2017
• Published: 09/07/2017
• Est. Priority Date: 03/31/2008
• Status: Active Grant

• Current Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Original Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Inventors: Turner, Stephen, Kuse, Ron, Kearns, Gregory, Monadgemi, Pezhman, Foquet, Mathieu, Martinez, Drew
• Granted Patent: US 10,562,766 B2
• Time in Patent Office: Days
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00509 Microcolumns
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0896 Nanoscaled
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites

Abstract

Methods, compositions and arrays for non-random loading of single analyte molecules into array structures are provided. Arrays of confined regions are produced wherein each confined region comprises a single island within the confined region. The island can be selectively functionalized with a coupling agent to couple a single molecule of interest within the confined region.

Claims

1. An array of nanoscale apertures comprising:

a transparent substrate having a cladding layer disposed on its surface, the cladding layer having a plurality of nanoscale apertures extending therethrough;

each nanoscale aperture having walls and a base, wherein an isolation layer is on the walls and on a portion of the base of the nanoscale aperture;

wherein a portion of the base of the nanoscale aperture comprises an island of substrate surrounded by isolation layer.

2. The array of claim 1 wherein the substrate comprises a silica based material.

3. The array of claim 1 wherein the substrate comprises a fused silica.

4. The array of claim 1 wherein the substrate comprises more than 10,000 nanoscale apertures.

5. The array of claim 1 wherein the nano scale apertures comprise holes having a circular lateral profile.

6. The array of claim 1 wherein the apertures have a cross sectional dimension between 1 nm and 500 nm.

7. The array of claim 1 wherein at least some of the plurality of nanoscale apertures comprise a single molecule of interest.

8. The array of claim 7 wherein the single molecule of interest comprises an enzyme.

9. The array of claim 1 wherein at least some of the plurality of nanoscale apertures comprise a single active polymerase enzyme, a single active template nucleic acid, or a single active primer attached to the island.

10-35. -35. (canceled)

36. A method for forming an island of substrate surface within a nanoscale aperture comprising:

providing a substrate having a cladding layer on top, the cladding layer having a plurality of nanoscale apertures extending therethrough;

conformally depositing an isolation layer on the cladding layer and exposed portions of the substrate;

conformally depositing a sacrificial layer onto the top of the isolation layer;

directionally etching the sacrificial layer such that the sacrificial layer remains on the walls of the nanoscale apertures, and the sacrificial layer is removed from the region of the nanoscale aperture between the sacrificial layer on the walls, exposing a portion of the isolation layer within the nanoscale aperture;

etching the portion of the isolation layer within the nanoscale aperture to expose a portion of the substrate; and

removing the sacrificial layer to produce a structure having an island of exposed substrate surface surrounded by isolation layer within each nanoscale aperture.

37. The method of claim 36 wherein the isolation layer is deposited by atomic layer deposition (ALD).

38. The method of claim 36 wherein the isolation layer comprises a metal oxide.

39. The method of claim 38 wherein the isolation layer comprises alumina.

40. The method of claim 36 wherein the substrate comprises a transparent material.

41. The method of claim 40 wherein the substrate comprises a silica based material.

45. The method of claim 36 wherein the sacrificial layer comprises silicon or germanium or silicon and germanium.

46. The method of claim 36 wherein the nanoscale apertures comprise holes having a circular lateral profile.

47. The method of claim 36 wherein the substrate comprises more than 10,000 nanoscale apertures.

48. The method of claim 36 further comprising binding a polymerase enzyme, template nucleic acid, or primer selectively onto the island.

42-44. -44. (canceled)

49. A method for forming an island of substrate surface within a nanoscale aperture comprising:

depositing an isolation layer onto a transparent substrate;

forming nanopits in the isolation layer that extend to the substrate surface;

depositing, exposing, and developing a resist to form a pillar of resist on top of and extending over each nanopit;

depositing a cladding layer such that the cladding layer covers the pillars of resist and the exposed regions of isolation layer; and

removing the resist resulting in lift-off of the portion of the cladding layer covering the pillars of resist, thereby forming a nanoscale apertures in the cladding layer, each having a nanopit at its base surrounded by isolation layer.

50-94. -94. (canceled)