Sample analysis method

• US 9,535,033 B2
• Filed: 08/02/2013
• Issued: 01/03/2017
• Est. Priority Date: 08/17/2012
• Status: Active Grant

• Current Assignee: Osaka University
• Original Assignee: Quantum Biosystems Inc.
• Inventors: Kawai, Tomoji, Ohshiro, Takahito, Taniguchi, Masateru
• Primary Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US14/421,809
• Publication Number:
  US 20150219593A1
• Time in Patent Office: 1,250 Days
• US Class Current: 1/1
• CPC Class Codes: G01N 15/1031 by measuring electrical or magnetic effects
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 27/3275 Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
• CPC Class Codes: G01N 27/447 using electrophoresis
• CPC Class Codes: G01N 27/4473 by electric means
• CPC Class Codes: G01N 33/48721 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

Abstract

The analysis method allows analysis of samples with high sensitivity, irrespective of interelectrode distance. The method includes: a step of applying a voltage between a first electrode pair such that an electric field is formed in a direction intersecting a migration direction of a sample; a step of placing a solution, including an electrochemically active molecule that produces a redox reaction at the electrode pair, between the first electrode pair; a step of causing the sample to migrate; and a step of measuring an amount of change in current flow between the first electrode pair.

Claims

1. A sample analysis method, comprising:

(a) applying a voltage between a first electrode pair, which is formed so as to sandwich a migration pathway of a sample such that an electric field is formed in a direction intersecting a migration direction of the sample;

(b) causing a first current, which arises from a redox reaction, to flow to the first electrode pair by placing, within the migration pathway interposed between the first electrode pair, a solution including an electrochemically active molecule that produces the redox reaction at the first electrode pair;

(c) causing the sample to migrate along the migration pathway interposed between the first electrode pair; and

(d) measuring an amount of change in the first current produced by migration of the sample.

2. The sample analysis method according to claim 1, further comprising calculating a volume of the sample from the measured amount of change in the first current, based on a correlation between a known volume of a reference sample and an amount of change in the first current, or an amount of change in a physical quantity according to the first current for the reference sample.

3. The sample analysis method according to claim 1, further comprising calculating a quantity of charge of the sample from the measured amount of change in the first current, based on a correlation between a known charge quantity of a reference sample and an amount of change in the first current, or an amount of change in a physical quantity according to the first current for the reference sample.

4. The sample analysis method according to claim 1, wherein:

(a) further comprises applying a voltage between a second electrode pair, which is formed so as to sandwich the migration pathway of the sample such that an electric field is formed in a direction substantially parallel to the migration direction of the sample;

(b) further comprises causing a second current to flow to the second electrode pair, wherein said second current arises from migration of ions contained in the solution along the migration direction of the sample through the migration pathway interposed between the first electrode pair; and

(d) further comprises measuring an amount of change in the second current produced by the migration of the sample.

5. The sample analysis method according to claim 4, further comprising calculating a volume of the sample from the measured amount of change in the second current, based on a correlation between a known volume of a reference sample and an amount of change in the second current, or an amount of change in a physical quantity according to the second current for the reference sample.

6. The sample analysis method according to claim 4, wherein:

the first electrode pair comprises gold electrodes or platinum electrodes;

the second electrode pair comprises silver/silver chloride electrodes; and

the ions comprise chloride ions.

7. The sample analysis method according to claim 6, wherein the chloride ions have a concentration of from about 1 mM to about 1 M.

8. The sample analysis method according to claim 4, wherein a distance between an anode and a cathode of the second electrode pair is less than about 100 micrometers.

9. The sample analysis method according to claim 1, wherein the electrochemically active molecule comprises a metal complex, an organometallic complex, or an organic molecule.

10. The sample analysis method according to claim 9, wherein the electrochemically active molecule comprises a potassium hexacyanoferrate complex, a hexamine ruthenium complex chloride, or hydroxyferrocene.

11. The sample analysis method according to claim 9, wherein the electrochemically active molecule causes a redox reaction when a voltage of −

1V to 1V is applied.

12. The sample analysis method according to claim 1, wherein a distance between an anode and a cathode of the first electrode pair is at least 2 nm.

13. The sample analysis method according to claim 1, wherein the direction of the electric field and the migration direction of the sample intersect at an angle of from about 45°

to about 90°

.

14. The sample analysis method according to claim 1, wherein the sample is spherical, and wherein a distance between an anode and a cathode of the first electrode pair is less than about 100 times a diameter of the sample.

15. The sample analysis method according to claim 1, wherein the sample comprises nucleic acids or amino acids.