Two-chamber dual-pore device

Patent metadata

Field	Value
Publication number	US-9696277-B2
Application number	US-201214358001-A
Country	US
Kind code	B2
Filing date	Nov 13, 2012
Priority date	Nov 14, 2011
Publication date	Jul 4, 2017
Grant date	Jul 4, 2017

Abstract

Provided is a device comprising a channel through and defined by a plurality of layers surrounding the channel, the channel connecting a first and a second chambers separated by the plurality of layers, wherein the plurality of layers comprise a first layer, a second layer; and a conductive middle layer disposed between the first and second layers, wherein the channel comprises (a) a first region defined by the first layer, denoted as an inlet, that is about 0.5 nm to about 100 nm in diameter and (b) a second region defined by the second layer, denoted as an outlet, wherein the inlet and the outlet are about 10 nm to about 1000 nm apart from each other, and wherein the first and second chambers and the middle layer are connected to a power supply. Also provided are methods of preparing and using the device, in particular for nucleic acid sequencing.

First claim

The invention claimed is:

1. A device comprising a channel through and defined by a plurality of layers surrounding the channel, the channel connecting a first chamber and a second chamber separated by the plurality of layers, wherein the plurality of layers comprise:
a first layer;
a second layer; and
a conductive middle layer disposed between the first and second layers,

wherein the channel comprises (a) an inlet defined by the first layer that is between 0.5 nm and 100 nm in diameter and (b) an outlet defined by the second layer, wherein the inlet and the outlet are between 10 nm and 1000 nm apart from each other; and wherein the first and second chambers and the middle layer are connected to a power supply; wherein the device further comprises at least one sensor capable of identifying a polymer or an individual component of a polymer during movement of the polymer through the inlet and outlet, wherein the sensor measures an ionic current through the inlet.

2. The device of claim 1 wherein the inlet has a depth of from 0.1 to 100 nm.

3. The device of claim 1 wherein the inlet has a diameter of 1 nm to 20 nm.

4. The device of claim 1 wherein the outlet has a diameter of 1 nm to 100 nm.

5. The device of claim 1 wherein the outlet has a depth of 0.1 nm to 100 nm.

6. The device of claim 1 wherein the device further comprises an additional layer wherein the additional layer comprises a dielectric material.

7. The device of claim 6 wherein the additional layer is 1 nm to 50 nm in thickness.

8. The device of claim 1 wherein the first layer and/or the second layer comprises a metallic material and wherein the device further comprises insulating material between the first layer and the middle layer and/or between the middle layer and the second layer.

9. The device of claim 8 wherein the middle layer comprises a metal selected from chrome, aluminum, platinum, and gold.

10. The device of claim 1 wherein the power supply is configured to provide a first voltage V 1 between the first chamber and the middle layer, and a second voltage V 2 between the middle layer and the second chamber, and wherein the first voltage and the second voltage are independently adjustable.

11. The device of claim 10 wherein the power supply comprises a voltage-clamp system to generate each of the first and second voltages.

12. The device of claim 10 wherein the middle layer is adjusted to be ground relative to the two voltages.

13. The device of claim 1 , wherein the sensor is configured to identify the polymer or the individual component by measuring a current, a voltage, pH, an optical feature, or residence time associated with the polymer or one or more components of the polymer.

14. The device of claim 1 , wherein the sensor is configured to form a tunnel gap allowing the polymer to pass through the tunnel gap when the polymer is loaded within the inlet and the outlet.

15. A method for controlling the movement of a charged polymer through a channel, comprising the steps of:
(a) loading a sample comprising a charged polymer in one of a first chamber or a second chamber of a device,
wherein the device comprises a channel through and defined by a plurality of layers surrounding the channel, the channel connecting a first chamber and a second chamber separated by the plurality of layers, wherein the plurality of layers comprise:
a first layer;
a second layer; and
a conductive middle layer disposed between the first and second layers,

wherein the channel comprises (a) an inlet defined by the first layer that is between 0.5 nm and 100 nm in diameter and (b) an outlet defined by the second layer,
wherein the inlet and the outlet are between 10 nm and 1000 nm apart from each other; wherein the first and second chambers and the middle layer are connected to a power supply; and
wherein the device is connected to a voltage-clamp or patch-clamp system for providing a first voltage between the first chamber and the middle layer, and a second voltage between the middle layer and the second chamber;

(b) setting an initial first voltage V 1 and an initial second voltage V 2 so that the polymer moves through the chambers, thereby locating the polymer across both the inlet and outlet; and
(c) adjusting the first voltage and the second voltage so that both voltages generate force to pull the charged polymer away from the middle layer, wherein the two voltages are different in magnitude, under controlled conditions, so that the charged polymer moves through the channel and both the inlet and the outlet in one direction and in a controlled manner.

16. The method of claim 15 wherein the adjusted first voltage and second voltage at step (c) are 10 times to 10,000 times as high, in magnitude, as the difference between the two voltages.

17. A device comprising a channel through and defined by a plurality of layers surrounding the channel, the channel connecting a first chamber and a second chamber separated by the plurality of layers, wherein the plurality of layers comprise:
a first layer;
a second layer; and
a conductive middle layer disposed between the first and second layers,
wherein the channel comprises (a) an inlet defined by the first layer that is between 0.5 nm and 100 nm in diameter and (b) an outlet defined by the second layer, wherein the inlet and the outlet are between 10 nm and 1000 nm apart from each other, wherein the outlet has a diameter greater than the diameter of the inlet; and wherein the first and second chambers and the middle layer are connected to a power supply.

18. The device of claim 17 , further comprising at least one sensor capable of identifying a polymer or an individual component of a polymer during movement of the polymer through the inlet and outlet, wherein the sensor is configured to identify the polymer or the individual component by measuring a current, a voltage, pH, an optical feature, or residence time associated with the polymer or one or more components of the polymer.

Assignees

• Univ California
• Univ Brigham Young

Inventors

• Dunbar William
• Schmidt Holger
• Hawkins Aaron

Classifications

• G01N33/48721 Primary

  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

• B82Y15/00

  Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors · CPC title

• G01N27/44791 Primary

  Microapparatus (sample containers with integrated microfluidic structures B01L3/5027) · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title

• C12Q2565/631

  being a biochannel or pore · CPC title