Systems and methods for forming a nanopore in a lipid bilayer

Patent metadata

Field	Value
Publication number	US-10456993-B2
Application number	US-201414335616-A
Country	US
Kind code	B2
Filing date	Jul 18, 2014
Priority date	Feb 8, 2010
Publication date	Oct 29, 2019
Grant date	Oct 29, 2019

Abstract

Techniques for forming a nanopore in a lipid bilayer are described herein. In one example, an agitation stimulus level such as an electrical agitation stimulus is applied to a lipid bilayer wherein the agitation stimulus level tends to facilitate the formation of nanopores in the lipid bilayer. In some embodiments, a change in an electrical property of the lipid bilayer resulting from the formation of the nanopore in the lipid bilayer is detected, and a nanopore has formed in the lipid bilayer is determined based on the detected change in the lipid bilayer electrical property.

First claim

What is claimed is:

1. A method of forming a nanopore in a lipid bilayer including:
determining by a processor that a lipid bilayer is formed in a nanopore cell based on a first measurement from a sensing circuit;
in response to the determination that the lipid bilayer is formed and after nanopore forming molecules are deposited on the lipid bilayer, controlling by the processor a source to apply an agitation stimulus level to the lipid bilayer for up to one second, causing a disruption in the lipid bilayer that tends to facilitate the formation of nanopores in the lipid bilayer; and
controlling by the processor the source to apply a checking stimulus level following the agitation stimulus level to the lipid bilayer and determining that a nanopore has been formed based on a second measurement received from the sensing circuit sensing in response to the checking stimulus level, wherein an absolute magnitude of the agitation stimulus level is greater than an absolute magnitude of the checking stimulus level.

2. The method of claim 1 , wherein the agitation stimulus is selected from the group consisting of: electrical stimulus, mechanical stimulus, sound stimulus, chemical light stimulus, and thermal stimulus.

3. The method of claim 1 wherein the agitation stimulus comprises a voltage (V) level.

4. The method of claim 1 , wherein the agitation stimulus level tends to facilitate the insertion of α-hemolysin nanopore in the lipid bilayer.

5. The method of claim 1 , further comprising:
detecting by the sensing circuit a change in an electrical property of the lipid bilayer resulting from the formation of a nanopore in the lipid bilayer;
determining by the processor that a nanopore has formed in the lipid bilayer based on the detected change in the lipid bilayer electrical property.

6. The method of claim 5 , wherein one or more steps are automated.

7. The method of claim 5 , wherein detecting a change in the lipid bilayer electrical property comprises detecting a change in a resistance of the lipid bilayer.

8. The method of claim 5 , wherein determining that a nanopore is formed includes determining a number of nanopores formed based on a size of change in the lipid bilayer electrical property.

9. The method of claim 8 , further including controlling by the processor the source to apply an erasing electrical stimulus to erase the lipid bilayer when it is determined that more than one nanopore is formed in the lipid bilayer.

10. The method of claim 8 , further including controlling by the processor the source to apply another agitation electrical stimulus level to a lipid bilayer wherein the another agitation electrical stimulus level tends to facilitate the formation of nanopores in the lipid bilayer when it is determined that no nanopore is formed in the lipid bilayer.

11. The method of claim 5 , wherein detecting a change in the electrical property of the bilayer resulting from the formation of a nanopore in the lipid bilayer includes detecting a decrease in resistance of the bilayer.

12. The method of claim 1 , wherein the lipid bilayer is formed on a substantially planar lipid bilayer compatible solid surface.

13. The method of claim 1 , wherein the lipid bilayer is formed on a substantially planar hydrophilic solid surface.

14. The method of claim 1 , wherein the lipid bilayer is formed on a substantially planar electrode surface.

15. The method of claim 1 , wherein the nanopore cell is one of a plurality of nanopore cells in a nanopore array.

16. The method of claim 15 , wherein each of the plurality of nanopore cells is individually addressable.

17. The method of claim 15 , wherein each of the plurality of nanopore cells is individually controllable.

Assignees

• Genia Tech Inc
• Roche Sequencing Solutions Inc

Inventors

• Chen Roger J A
• Davis Randy

Classifications

• B82Y40/00

  Manufacture or treatment of nanostructures · CPC title

• G01N33/48721

  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 title

• Y10S977/797

  Lipid particle · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title

• B29C67/20 Primary

  for porous or cellular articles, e.g. of foam plastics, coarse-pored {(chemical aspects of working up macro-molecular substances to porous or cellular articles C08J9/00)} · CPC title