Device for the analysis of nucleic acid molecules

Patent metadata

Field	Value
Publication number	US-10933416-B2
Application number	US-201615571679-A
Country	US
Kind code	B2
Filing date	May 6, 2016
Priority date	May 7, 2015
Publication date	Mar 2, 2021
Grant date	Mar 2, 2021

Abstract

The invention proposes a device for analyzing nucleic acid molecules (M), comprising: —a bead (20), on which one molecule can be anchored at one end, —a surface (520), on which the molecule can be anchored at the other end, —an actuator (30), adapted to cause the bead to move relative to said surface in one direction, —a sensor (50), adapted to measure a distance between the bead and the surface, the device further comprising a well (11), having an axis (X-X) extending along the direction of motion of the bead and a bottom (110) formed by said surface, said well being filled with electrically conductive solution (40), and receiving the bead, the sensor being adapted to measure an impedance of the well, depending on a distance between the bead and the surface, to determine, the distance between the bead and the surface.

First claim

The invention claimed is:

1. Device for analyzing nucleic acid molecules, comprising:
a well, comprising:
(i) a bottom surface and a top surface opposite the bottom surface,
(ii) an axis extending between the bottom surface and the top surface,
(iii) a nucleic acid molecule anchored at a first end to the bottom surface;
(iv) an electrically conductive solution, filling at least part of the well; wherein a cross-sectional area of the well perpendicular to the axis increases from the bottom surface to the top surface of the well; and

a bead disposed within the well, the bead anchored at a second end of the nucleic acid molecule, the second end opposite the first end;
an actuator configured to cause the bead to move relative to the bottom surface along the axis of the well; and
a sensor configured to measure an impedance of the well, the impedance depending on a distance between the bead and the surface, such that, from the measured impedance, the distance between the bead and the surface is determined.

2. The device according to claim 1 , wherein the sensor comprises:
a main electrode, positioned on the top surface of the well, in contact with the electrically conductive solution, the electrode being submitted to a known potential,
a secondary electrode adjacent the bottom surface of the well,
, and
an electronic circuit, configured to measure a current flowing between the electrodes.

3. The device according to claim 2 , wherein the electronic circuit comprises:
a current to voltage amplifier connected to the secondary electrode,
a voltmeter adapted to measure an output voltage of the current to voltage amplifier, and
a computing circuit adapted to compute an impedance of the well from the measured voltage.

4. The device according to claim 1 , wherein the area of the cross-section at the bottom of the well is greater than the area of the largest cross-section of the bead.

5. The device according to claim 1 , wherein the well has a shape of a truncated cone.

6. The device according to claim 1 , wherein the cross-sectional area of the well increases in a linear relationship with respect to the distance from the bottom surface of the well.

7. The device according to claim 6 , wherein the cross-section of the well is circular at the bottom surface and increases linearly in a direction orthogonal to the axis of the well with the distance from the bottom surface of the well.

8. The device according to claim 1 , wherein the actuator comprises at least one magnet mounted to be displaceable along the direction of the axis of the well, and the bead is made of a paramagnetic material, and the bead is interposed between the bottom surface of the well and the at least one magnet.

9. The device according to claim 1 , further comprising a plurality of identical beads and a plurality of identical wells, each of the plurality of identical wells adapted to receive one bead.

10. The device according to claim 1 , further comprising:
a plurality of identical beads and a plurality of identical wells, each of the plurality of identical wells having one bead of the plurality of identical beds therein, each well of the plurality of identical wells provided with a sensor, the sensor comprising:
a main electrode, positioned on the top of the well and in contact with the electrically conductive solution, the electrode being submitted to a known potential,
a secondary electrode adjacent the bottom surface of the well, and
an electronic circuit, configured to measure a current flowing between the electrodes, the electronic circuit comprises a current to voltage amplifier, the current to voltage amplifier connected to the secondary electrode, and a computing circuit configured to simultaneously measure output voltages of the current to voltage amplifier and compute the impedance of the respective wells.

11. The device according to claim 1 , further comprising a plurality of identical beads and a plurality of identical wells, in which one bead of the plurality of identical beads is disposed, each of the identical wells adapted to receive one bead.

12. The device according to claim 11 , wherein:
the actuator comprises at least one magnet displaceably mounted such that the at least one magnet is displaceable along the axis of the well; and
the bead comprises a paramagnetic material and is interposed between the bottom surface of the well and the magnet.

13. The device according to claim 12 , wherein the at least one magnet comprises a plurality of bars, the plurality of bars made of a magnetic material, and each bar of the plurality is disposed on a top surface of the plate between two adjacent wells of the plurality of wells.

14. The device according to claim 13 , wherein each bar of the plurality of bars has a length less than 10 μm and has a diameter less than or equal to 1 μm.

15. The device according to claim 1 , wherein the well is formed as a depression in a plate of electrically insulating material.

16. The device according to claim 1 wherein the dimensions of the well are configured to keep a Johnson noise produced during measurement under a predetermined amount, the amount determined upon a desired solution of a signal measured from the well.

17. The device according to claim 1 wherein the dimensions of the well are configured to keep the signal to noise ratio at least 10 to 1.

18. The device according to claim 1 wherein the dimensions of the well are configured based on the current noise and bandwidth of a frequency.

Assignees

• Paris Sciences Et Lettres—Quartier Latin
• Centre Nat Rech Scient
• Univ Paris Diderot Paris 7
• Univ Sorbonne

Inventors

• Croquette Vincent
• Bensimon David
• Ouellet Jimmy
• Allemand Jean-François
• Andre Charles
• Hamilton Gordon

Classifications

• G01N33/5308

  for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites · CPC title

• C12Q1/6825 Primary

  Nucleic acid detection involving sensors · CPC title

• B01L2400/0436

  acoustic forces, e.g. surface acoustic waves [SAW] · CPC title

• B01L2400/043

  magnetic forces · CPC title

• B01L2300/16

  Surface properties and coatings · CPC title