Biopolymer optical analysis device and method

Patent metadata

Field	Value
Publication number	US-9562809-B2
Application number	US-201113812801-A
Country	US
Kind code	B2
Filing date	Jul 21, 2011
Priority date	Sep 29, 2010
Publication date	Feb 7, 2017
Grant date	Feb 7, 2017

Abstract

The present invention provides a device and method for analyzing the characteristics of a biopolymer with excellent mechanical stability, high spatial resolution and sensitivity using a simple device construction. Specifically, the Raman scattered light of a biopolymer is measured and the properties of monomer units forming the biopolymer are analyzed by using a biopolymer property analysis chip ( 100 a ) characterized by comprising: a solid substrate ( 110 ); at least one nanopore ( 120 ) disposed in the solid substrate ( 110 ); and one or more electrically conductive thin films ( 130 a, 130 b ) disposed on the solid substrate ( 110 ). The biopolymer property analysis chip ( 100 a ) is characterized in that the electrically conductive thin films ( 130 a, 130 b ) are disposed partially on the solid substrate ( 110 ) where the nanopore ( 120 ) is formed and a biopolymer which has penetrated into the nanopore ( 120 ) is caused to generate Raman scattered light by means of irradiation with external light.

First claim

The invention claimed is:

1. A biopolymer property analysis chip comprising:
a solid state substrate having a thin film made of an electric insulator material;
at least one nanopore formed in the solid state substrate; and
at least one electroconductive thin film disposed on the solid state substrate, wherein
the nanopore penetrates through the thickness of the thin film made of an electric insulator material,
the electroconductive thin film is disposed partially in contact with a circumference of the nanopore,
a biopolymer which has penetrated into the nanopore is caused to generate a Raman scattered light by irradiation with an external light, and
only a portion of the entire circumference of the nanopore has a plasmon resonance member structure,
the plasmon resonance member structure is an electroconductive thin film consisting of a single layer,
the plasmon resonance member structure is not in contact with another conductive material,
the electroconductive thin film is disposed in a middle depth of the nanopore along its central axis in the solid state substrate,
the thin film of the electric insulator material including a first film and a second film, and
the electroconductive thin film is sandwiched between the first film and the second film, such that a top surface and a bottom surface of the electroconductive thin film are not exposed.

2. The biopolymer property analysis chip defined in claim 1 , wherein the external light is irradiated on the electroconductive thin film so as to generate a near field at an edge facing the opening of the nanopore, and the biopolymer which has penetrated into the nanopore is caused to generate a Raman scattered light.

3. The biopolymer property analysis chip defined in claim 1 , wherein the electroconductive thin film has an acute-angled edge, the acute-angled edge being disposed facing the opening of the nanopore.

4. The biopolymer property analysis chip defined in claim 1 , wherein at least the two electroconductive thin films are formed, at least the two electroconductive thin films being disposed so as to sandwich the opening of the nanopore facing each other.

5. The biopolymer property analysis chip defined in claim 1 , wherein the electroconductive thin films are made of metal.

6. The biopolymer property analysis chip defined in claim 1 , wherein the electroconductive thin films are made of graphite.

7. The biopolymer property analysis chip defined in claim 1 , wherein the thickness of the electroconductive thin films is 0.1 nm to 10 nm.

8. The biopolymer analysis chip defined in claim 1 , wherein the solid state substrate has a thin film part which substantially transmits light and the nanopore is formed on the thin film part.

9. The biopolymer property analysis chip defined in claim 1 , wherein the electroconductive thin films are disposed on the surface of the solid state substrate.

10. The biopolymer property analysis chip defined in claim 1 , wherein the depth of the nanopore is three times or more that of monomer units composing biopolymer.

11. The biopolymer property analysis chip defined in claim 1 , wherein the biopolymer is selected from the group of nucleic acid, peptide nucleic acid, protein, sugar chain, and aptamer.

12. The biopolymer property analysis chip defined in claim 1 , wherein the biopolymer property analysis is to determine the base sequence of nucleic acid.

13. A biopolymer property analysis device comprising:
the biopolymer property analysis chip defined in claim 1 ;
a light source; and
a one-dimensional or two-dimensional detector with frame rate of 1 kHz or more, wherein
external light is irradiated on the analysis chip from the light source, and a Raman scattered light from the biopolymer at the analysis chip is detected using the detector.

14. The biopolymer property analysis device defined in claim 13 , further comprising frame buffer memory for recording the measured valued read out from the detector.

15. The biopolymer property analysis device defined in claim 13 , further comprising a detector having a photointensifier part as the detector.

16. The biopolymer property analysis device defined in claim 13 , further comprising a driving part configured to cause the monomers in a biopolymer to enter the nanopore one by one.

17. A method for analyzing the characteristics of a biopolymer comprising the steps of:
causing the biopolymer which has penetrated into the nanopore to generate a Raman scattered light by irradiation on a biopolymer property analysis chip defined in claim 1 , with external light and
analyzing the characteristics of the biopolymer based on the Raman scattering spectrum.

18. The method for analyzing the characteristics of a biopolymer defined in claim 17 , wherein the biopolymer is selected from the group of nucleic acid, peptide nucleic acid, protein, sugar chain, and aptamer.

19. The method for analyzing the characteristics of a biopolymer defined claim 17 , wherein the base sequence of nucleic acid is determined.

20. The method for analyzing the characteristics of a biopolymer defined in claim 17 , wherein the biopolymer is contained in a sample solution containing a second polymer, which cannot penetrate into the nanopore.

Assignees

• Ozawa Satoshi
• Anazawa Takashi
• Akahori Rena
• Takahashi Satoshi
• Ohura Takeshi
• Kiguchi Masashi
• Hitachi High Tech Corp

Inventors

• Ozawa Satoshi
• Anazawa Takashi
• Akahori Rena
• Takahashi Satoshi
• Ohura Takeshi
• Kiguchi Masashi

Classifications

• B82Y5/00 Primary

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• G01J3/0267 Primary

  Sample holders for colorimetry · CPC title

• B82Y15/00

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

• G01N21/658

  enhancement Raman, e.g. surface plasmons · CPC title