Substance-Information Acquisition Method Using Evanescent Light Beam, Substance-Information Measurement Apparatus, Base-Sequence Determination Method and Base-Sequence Determination Apparatus

• US 20100203504A1
• Filed: 06/20/2006
• Published: 08/12/2010
• Est. Priority Date: 07/07/2005
• Status: Abandoned Application

• Current Assignee: Sony Corporation (Sony Group Corp.)
• Original Assignee: Sony Corporation (Sony Group Corp.)
• Inventors: Kishii, Noriyuki, Katsumoto, Yoichi
• Application Number:
  US11/993,786
• Publication Number:
  US 20100203504A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 20/00 Nanooptics, e.g. quantum optics or photonic crystals
• CPC Class Codes: B82Y 35/00 Methods or apparatus for measurement or analysis of nanostructures
• CPC Class Codes: G01B 11/026 by measuring distance between sensor and object G01B11/0608 takes precedence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01Q 60/20 Fluorescence

Abstract

Information on the position and displacement of a detection-subject substance T can be presented by adoption an evanescent light illumination technology. An evanescent light beam PE has a property that the intensity thereof abruptly attenuates in accordance with an exponential function with increases in distance Z from a boundary surface S capable of generating the evanescent light beam PE. The detection-subject substance T fixed on the boundary surface S is excited by the evanescent light beam PE having the property described, generating the original fluorescent light beam f of the detection-subject substance T. On the basis of the intensity I of the fluorescent light beam f, information Z on the position of the detection-subject substance T and information Δz on the displacement of the detection-subject substance T can be acquired. In addition, by making use of the information Z and the information Δz, it is possible to obtain information on the structure of the detection-subject substance T, its structural change, a base sequence of a nucleic acid chain serving as the detection-subject substance T, the existence/non-existence of substance interactions and the state of progress of the interactions.

Claims

1. A substance-information acquisition method, a detection-subject substance is put in a boundary surface area capable of generating an evanescent light beam and information on the position of said detection-subject substance is acquired on the basis of the intensity of a fluorescent light beam generated by said detection-subject substance excited by said evanescent light beam as an original light beam of said detection-subject substance.

2. The substance-information acquisition method according to claim 1, wherein said detection-subject substance is fixed on said boundary surface area.

3. The substance-information acquisition method according to claim 1, information on a displacement of said detection-subject substance is obtained by acquiring fluorescence-intensity information before a change of a physical or chemical condition of a reaction field in which said detection-subject substance exists and fluorescence-intensity information after said change.

4. The substance-information acquisition method according to claim 3, wherein said information on a displacement is information on a quantity bringing about a change of the structure or shape of said detection-subject substance.

5. The substance-information acquisition method according to claim 1, the intensity of said fluorescent light beam generated by said detection-subject substance excited by said evanescent light beam is monitored in a real-time manner.

6. The substance-information acquisition method according to claim 1, wherein said information on the position of said detection-subject substance is nano-scale positional information.

7. The substance-information acquisition method according to claim 1, said information on the position of said detection-subject substance is determined by correcting data of a measured intensity of said fluorescent light beam on the basis of standard data acquired in advance.

14. The substance-information acquisition method according to claim 4, wherein

said detection-subject substance is a detection-subject nucleic acid chain fixed on said boundary surface area, and

information on a displacement caused by extension of said detection-subject nucleic acid chain is obtained.

8. A substance-information measurement apparatus comprising:

a light source;

a boundary surface capable of generating an evanescent light beam due to a light beam generated by said light source to hit said boundary surface;

a fluorescent light detection section for detecting the intensity of a fluorescent light beam generated by a detection-subject substance existing on said boundary surface as a substance excited by said evanescent light beam; and

an analysis section for automatically analyzing the position and/or displacement of said detection-subject substance in a reaction field on the basis of said intensity of said fluorescent light beam and an attenuation rate exhibited by said evanescent light beam as an attenuation rate having a correlation with a distance from said boundary surface.

9. The substance-information measurement apparatus according to claim 8, said substance-information measurement apparatus further provided with a reaction-field condition control section capable of changing a physical or chemical condition of said reaction field provided on a boundary-surface side opposite to a boundary-surface side, through which said light beam generated by said light source is propagating in the forward direction toward said boundary surface, as a field facing said boundary surface.

10. The substance-information measurement apparatus according to claim 9, wherein said reaction-field condition control section includes field applying means for applying an electric field to said reaction field.

15. The substance-information measurement apparatus according to claim 8, wherein

said detection-subject substance is a detection-subject nucleic acid chain fixed on said boundary surface area, and

a structure change based on a hybridization reaction between said detection-subject nucleic acid chain and a target nucleic acid chain as a structure change of said detection-subject is analyzed.

11. A base-sequence determination method for determining a base sequence of a single-strand nucleic acid chain or a double-strand nucleic acid chain by execution of procedures for:

letting a protein be held in advance in a boundary-surface area capable of generating an evanescent light beam as a protein having a structure, which changes when said protein is joined to a base of said single-strand nucleic acid chain or a base pair of said double-strand nucleic acid chain;

joining said protein to said single-strand nucleic acid chain or said double-strand nucleic acid chain;

continuously measuring the intensity of a fluorescent light beam, which is generated by said protein excited by said evanescent light beam as the original fluorescent light beam of said protein, along the time axis; and

automatically collating fluorescent-light intensity data obtained as a result of said continuous-measurement procedure with a fluorescent-light intensity database provided for said protein as a database reflecting correlations with types of said base or types of said base pair.

12. The base-sequence determination method according to claim 11 wherein the structure of said protein changes in recognition units each comprising a plurality of said bases or a plurality of said base pairs.

13. A base-sequence determination apparatus comprising:

a boundary surface for holding a protein having a structure, which changes when said protein is joined to a base of a single-strand nucleic acid chain or a base pair of a double-strand nucleic acid chain, after putting a fluorescent substance on said protein in advance as a mark and for generating an evanescent light beam;

a fluorescence-intensity detection section capable of continuously measuring the intensity of a fluorescent light beam generated by said fluorescent substance as the original fluorescent light beam of said protein excited by said evanescent light beam;

a database storage section used for storing a fluorescent-light intensity database provided for said protein as a database reflecting correlations with types of said base or types of said base pair; and

a data processing section for automatically collating fluorescent-light intensity data obtained as a result of said measurement carried out by said fluorescence-intensity detection section with said fluorescent-light intensity database in order to determine a base sequence of said single-strand nucleic acid chain or a base sequence of said double-strand pair of nucleic acid chain.