High-speed DNA sequencing with optically active nanopore

Patent metadata

Field	Value
Publication number	US-10330631-B2
Application number	US-201514815228-A
Country	US
Kind code	B2
Filing date	Jul 31, 2015
Priority date	Jul 31, 2015
Publication date	Jun 25, 2019
Grant date	Jun 25, 2019

Abstract

A nanoscale-sized pore positioned between two reservoirs may sequence biomolecules by detecting changes in the emitted light due to a change in charge of portions of the biomolecules as they pass through the pore such as affect an emission frequency of a quantum structure proximate to the pore opening. The nanopores may be fabricated using local droplet etching whose randomness is accommodated by lowering the droplet density to permit isolation of nanopores in tiles that may be adhered to an underlying supporting substrate having an aligned opening. The nanopore-tiles may be integrated with commonly applied glass chips and may be employed in microfluidic circuitry.

First claim

We claim:

1. An apparatus for a measurement of biomolecules comprising:
a separator having a nanopore providing a passage through the separator, the nanopore incorporating a nanoscale semiconductor element proximate and fixed with respect to the passage and adapted to emit light, such light as emitted from the nanoscale semiconductor element having a frequency dependent on a charge of a portion of a biomolecule passing through the nanopore, the nanoscale semiconductor element providing physical structure producing a quantum confinement of electrons dropping between a conduction band and valence band to change a frequency of the light emitted by those electrons as a function of field induced interaction with the biomolecules;
a reservoir system holding a fluid on opposite sides of the separator to provide a flow of biomolecules through the nanopore from one side of the separator to the other;
a spectrometer receiving emitted light from the nanoscale semiconductor element to measure frequency of that light with flow of biomolecules through the nanopore; and
an electronic computer communicating with the spectrometer and executing a program to relate light frequency measured by the spectrometer to the structure of the portion of the biomolecules thereby providing a sequencing of a biomolecule structure as it passes through the nanopore.

2. The apparatus of claim 1 wherein the nanoscale semiconductor element is a donut concentric with the nanopore and bounded by different materials on nanoscale dimensions to provide a structure exhibiting quantum confinement effects.

3. The apparatus of claim 2 wherein the different materials are semiconductor materials.

4. The apparatus of claim 3 wherein the nanoscale semiconductor element and different materials are selected from group III/V semiconductors; group II/V semiconductors, and strained silicon and/or germanium.

5. The apparatus of claim 4 wherein the group III/V semiconductors are selected from the group consisting of gallium arsenide, aluminum gallium arsenide, and indium arsenide.

6. The apparatus of claim 1 wherein the separator is a solid material substantially unbroken outside of the nanopore over an area contacting fluid of the reservoir structure.

7. The apparatus of claim 1 wherein the separator is a membrane holding the nanopore and adhered to a substrate of different material, the substrate having an aperture aligned with the nanopore.

8. The apparatus of claim 1 wherein the reservoir system includes electrodes communicating with reservoir fluids to provide for an ionic flow from one side of the separator to the other.

9. The apparatus of claim 1 wherein the nanopore is substantially circular in cross-section.

10. The apparatus of claim 1 wherein the nanopore is noncircular in cross-section and sized to control an orientation of the biomolecules that are non-circular in cross-section as they pass through the nanopore.

11. The apparatus of claim 1 including an output device providing a human readable output indicating a sequence of the biomolecules.

12. An apparatus for a measurement of biomolecules comprising:
a separator having a nanopore providing a passage through the separator, the nanopore incorporating a nanoscale semiconductor element proximate to the passage and adapted to emit light with a frequency dependent on a charge of a portion of a biomolecule passing through the nanopore;
a reservoir system holding a fluid on opposite sides of the separator to provide a flow of biomolecules through the nanopore from one side of the separator to the other;
a spectrometer receiving emitted light from the nanoscale semiconductor element to measure frequency of that light with flow of biomolecules through the nanopore; and
an electronic computer communicating with the spectrometer and executing a program to relate light frequency measured by the spectrometer to the structure of the portion of the biomolecules thereby providing a sequencing of a biomolecule structure as it passes through the nanopore;
wherein the nanoscale semiconductor element is a donut concentric with the nanopore and hounded by different materials on nanoscale dimensions to provide a structure exhibiting quantum confinement effects; and
wherein the nanopore has a dimension of less than 1000 nanometers and wherein the doughnut extends less than 10 nanometers inward from the nanopore passage measured in a plane normal to an axis of the nanopore through the separator.

13. An apparatus for a measurement of biomolecules comprising:
a separator having a nanopore providing a passage through the separator, the nanopore incorporating a nanoscale semiconductor element proximate to the passage and adapted to emit light with a frequency dependent on a charge of a portion of a biomolecule passing through the nanopore;
a reservoir system holding a fluid on opposite sides of the separator to provide a flow of biomolecules through the nanopore from one side of the separator to the other;
a spectrometer receiving emitted light from the nanoscale semiconductor element to measure frequency of that light with flow of biomolecules through the nanopore; and
an electronic computer communicating with the spectrometer and executing a program to relate light frequency measured by the spectrometer to the structure of the portion of the biomolecules thereby providing a sequencing of a biomolecule structure as it passes through the nanopore;
further including a light source for providing stimulating energy to the nanoscale semiconductor element to promote an emission of light from the nanoscale semiconductor element.

14. A method for characterizing biomolecules comprising the steps of:
(a) passing the biomolecules from a first to a second reservoir through a nanopore providing a nanoscale semiconductor element proximate and fixed with respect and to the nanopore to emit light, such light, as emitted from the nanoscale semiconductor element, having a frequency dependent on a charge of a portion of a biomolecule passing through the nanopore, the nanoscale semiconductor element having structure providing quantum confinement of electrons dropping between a conduction band and valence band to change a frequency of the light emitted by those electrons as a function of field induced interaction with the biomolecules;
(b) during a transit of the biomolecules through the nanopore, applying a stimulating energy to the nanoscale semiconductor element to promote an emission of light therefrom;
(c) measuring a frequency of emitted light; and
(d) relating the frequency of emitted light over time to a sequence of structures of the biomolecules passing through the nanopore to provide a sequencing of the biomolecules.

15. A method of manufacturing separators for isolating reservoirs of liquid across at least one optically active nanopore, the method comprising the steps of:
(A) fabricating a matrix material on a sacrificial layer supported by a first substrate;
(B) subjecting the matrix material to local droplet etching in which metal droplets erode nanoscale holes through the matrix material to the sacrificial layer;
(C) preparing a second substrate with a plurality of apertures larger than the nanoscale holes, wherein the second substrate is thicker than the matrix material;
(D) removing the matrix material from the sacrificial layer and adhering it to the second substrate so that at least one nanopore aligns with at least one aperture; and
(E) dividing the adhered matrix material and second substrate to provide at least one separator element including a continuous passage through a nanoscale hole and aperture.

Assignees

• Wisconsin Alumni Res Found

Inventors

• Blick Robert H
• Gwozdz Paul H
• Bhat Abhishek
• Hansen Wolfgang
• Heyn Christian

Classifications

• B82Y15/00

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

• G01N21/6408

  with measurement of decay time, time resolved fluorescence · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · 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

• G01N2030/8827

  involving nucleic acids · CPC title