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WO2007060988A1 - Procede et dispositif de detection d'une substance a l'etat de trace par effet raman exalte de surface - Google Patents

Procede et dispositif de detection d'une substance a l'etat de trace par effet raman exalte de surface Download PDF

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Publication number
WO2007060988A1
WO2007060988A1 PCT/JP2006/323313 JP2006323313W WO2007060988A1 WO 2007060988 A1 WO2007060988 A1 WO 2007060988A1 JP 2006323313 W JP2006323313 W JP 2006323313W WO 2007060988 A1 WO2007060988 A1 WO 2007060988A1
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WIPO (PCT)
Prior art keywords
mixture
analyte
solid
metal
trace
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PCT/JP2006/323313
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English (en)
Japanese (ja)
Inventor
Tomohiro Marui
Akito Sekiguchi
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Intellectual Property Bank Corp
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Intellectual Property Bank Corp
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/65—Raman scattering
    • G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons

Definitions

  • the present invention is a method and apparatus for detecting trace substances by surface enhanced Raman scattering (SERS).
  • SERS surface enhanced Raman scattering
  • the Raman scattering enhancement effect in the vicinity of the metal particles in the dried body in which the colloid in which the metal particles are suspended is dried is used.
  • the present invention also utilizes relatively clean metal surfaces that induce surface plasmon polaritons with irradiated light energy.
  • a metal surface that has the effect of further enhancing surface enhanced Raman scattering (SERS) in the vicinity of the metal surface is provisionally called "a metal surface having a plasmon mirror effect", and the metal surface is used.
  • a metal surface having a plasmon mirror effect is preferably a clean surface of gold or a clean surface of silver.
  • Patent Document 1 discloses a simple method of preparing a substrate exhibiting high SERS activity by dispersing the SERS substrate in a liquid and fixing it on a solid surface.
  • a device for self-assembly (self-alignment) of nanoparticle groups to obtain strong SERS activity has also been studied.
  • a method of sculpting a group of fine metal particles which has been self-assembled at an interface according to a method of manufacturing LB film (Langmuir-Blodgett film) to form a SERS substrate in which metal particles are aligned is known (see FIG. 3 and Patent Document 3).
  • Patent Document 2 also contains metal fine particles.
  • a flat plate immersed in a solution is repeatedly pulled up at a predetermined speed to form a SERS substrate on which particles are periodically aligned.
  • a colloid in which precious metal nanoparticles are suspended is used as a SERS substrate (not referred to as a substrate because it is a liquid), to which an analyte is mixed in a liquid phase. It is also known to detect surface-enhanced Raman scattering for differential detection of analytes
  • Colloidal SERS substrates are easier to prepare than solid phase SERS substrates, but have the disadvantage that SERS activity disappears in the state of separation and precipitation immediately. Therefore, methods of stabilizing the SERS substrate for a long time with a clay-like substance such as smectite (see Patent Document 4) and the like have been studied.
  • capillary action capillary
  • colloidal SERS substrates in capillarity (capillary) and analyze the SERS Raman signal to discriminate trace substances.
  • Patent Document 8 discloses a method and apparatus for detecting a trace substance using the “Brazmon mirror effect”.
  • the Plasmon mirror effect is a self-assembly (self-alignment) of a group of nanoparticles in the vicinity of a relatively clean metal surface that induces surface plasmon polariton at the same time as the irradiated light energy, thereby obtaining stronger SERS activity. is there.
  • the conditions for obtaining the plasmon mirror effect and the mechanism thereof are not clear, it is considered important to self-assemble the nanoparticle group in the vicinity of a relatively clean metal surface (Patent Document 8) reference).
  • Patent Document 1 Japanese Patent Application Laid-Open No. 2005-077362 "Method for producing surface-enhanced Raman scattering active substrate” (School corporation Keio University)
  • Patent Document 2 Japanese Patent Application Laid-Open No. 2004-170334 "Raman Scattering Measurement Sensor and Method of Manufacturing the Same"
  • Patent Document 3 JP-A-2005-233637 "Raman spectroscopy analysis using gold nanorod thin film” (Japan Science and Technology Agency)
  • Patent Document 4 Japanese Patent Application Laid-Open No. 2004-205435 "Analytical method of a substance to be analyzed for binding ability which does not require a labeling dye and an analysis kit used therefor" (Fukuoka Takao)
  • Patent Document 5 Japanese Patent Application Laid-Open No. 2005-219184 "Method for forming metal nanotriangular columnar structure array substrate for single molecule Raman spectroscopy and single molecule analysis method using it" (National Institute of Advanced Industrial Science and Technology)
  • Patent Document 6 Patent No. 3462339 Publication No. "Detector for capillary electrophoresis” (Nippon Telegraph and Telephone Corporation)
  • Patent Document 7 Japanese Published Patent Application No. 2005-507500 "Detection of Microfluid by Surface-Enhanced Resonance Raman Scattering" (University Obstrastrice et al.)
  • Patent Document 8 International Publication WO2005 114298 "OPTICAL SENSOR WITH LAYERED P LASMON STRUCTURE FOR ENHANCED DETECTION OF CHEMICAL GROUPS BY SERS J VP HODLINGS. LLC
  • Patent Document 9 U.S. Pat. No. 6,149,868 "Surface enhanced raman scattering from meta
  • Non-patent literature 1 Kotaro Ashikawa (Tokyo Inst. Of Tech.), Ryota Mitsui "Biosensing using localized plasmon resonance” Applied Physics, 72nd, 12th, p. 1541-1544 (2003)
  • Non-patent literature 2 Takamoto Okamoto (RIKEN) "Research on metal nanoparticle interaction and bio-sensors” Grant-in-Aid for Scientific Research in fiscal 2002 (Basic research C) "Localization of surface brazmon and its application Research on “research research report”
  • Non-patent literature 3 Keir R, Igata E, Arundell M, Smith WE, Graham D, McHugh C, Coop er JM. TSERRS. In situ substrate formation and improved detection using microfluidic sJ Anal Chem. 2002 Apr 1; 74 (7) 1503-8.
  • Patent Document 4 Jacquitta K. Daniels and George S. humanov (Clemson University) "Na noparticle-Mirror Sandwich Substrates for Surface-Enhanced Raman Scattering" J. Phys. Chem. B, 109 (38), 17936-17942, 2005.
  • Non-patent literature 5 Orendorff, CJ Gole, A. Sau, TK Murphy, CJ (University of South Carolina) “Surface-enhanced Raman spectroscopy of Self-assembled monolayers: S andwich architecture and nanoparticles shape dependency j Anal Chem. 2005 May 15 77 (10): 3261-6.
  • An object of the present invention is to propose a method and an apparatus which are more practical than the conventional SERS substrate and a method and apparatus for detecting a trace substance by the SERS substrate.
  • the present invention proposes a method and apparatus in which a colloidal SERS substrate is used without using a substrate in which metal microparticles are aligned in a solid phase, but separation and precipitation of the substrate is not a problem.
  • the present invention is a method for discriminating and detecting a microanalyte with surface-enhanced Raman scattered light obtained by irradiating a laser beam to a portion where the analyte and the metal fine particle are in proximity to each other.
  • This is a method of detecting trace substances by surface-enhanced Raman scattering, which includes the steps of irradiating laser light to the peripheral part of the dried mixture on a flat surface and observing Raman scattered light (see Fig. 1 and Fig. 6).
  • the fine metal particle colloid prepared beforehand is a clay-like substance such as smectite, and there is no need to stabilize the SERS substrate for a long period of time.
  • known metal fine particle colloids can be obtained by reducing metal salts such as citric acid reduction of silver salts such as silver nitrate, reduction of silver salts such as silver nitrate with borohydride (borane), and the like.
  • Borohydride reagent is water Sodium borohydride is often used.
  • the solid to be dried is preferably glass or polycarbonate. Conventionally, it has not been possible to dry and observe a liquid-phase mixture of metal particle colloid and analyte on a solid surface. In the present proposal, it is especially possible to prepare only ordinary metal fine particle colloids without carefully devising measures of stability, mix it with the analyte and dry it, and observe the dried body.
  • FIGS. 2 to 4 The conventional method is shown in FIGS. 2 to 4 and the method of the present invention contrasted with this is shown in FIG.
  • SS is a known SERS substrate
  • SB is a bottle containing a known SERS substrate (SERS vial)
  • SE is a solid flat surface of the present invention onto which a mixture of metal fine particle colloid and analyte is dropped
  • p is a micropipette Means for dropping liquid.
  • a flow chart of the method for detecting a trace substance according to the present invention is shown in FIG.
  • FIG. 5 schematically shows a stin-like dried body S obtained by the method of the present invention, and a massive structure A and a bowl-like structure D are formed on the peripheral portion R of S.
  • Figure 8 exemplifies an optical micrograph of a bowl-like structure D formed on the rim R of S and a massive structure A.
  • FIG. 8 shows a dried mixture of silver fine particle colloid having a particle size of 20 to 30 nm, which is produced by quannic acid reduction of silver nitrate, using the analyte as an oligonucleotide.
  • the number of bases of the oligonucleotide is approximately 1000.
  • FIG. 7 An example of data of a Raman spectrum by the proposed detection method using an oligonucleotide as the analyte is shown in FIG. 7.
  • DA is a Raman spectrum observation position observed by moving a portion A or D linearly and observed by laser irradiation
  • N is a portion where neither A nor D is moved linearly but measured laser irradiation L It is the observation position of the vector. In this way, an enhanced large Raman signal is observed at the A or D site, and discrimination detection of the analyte is possible.
  • the present invention utilizes a colloidal SERS substrate, but separation and precipitation of the substrate is not a problem. The Therefore, it is practical because it is sufficient to prepare only ordinary metal fine particle colloids which are not specially designed to stabilize the substrate.
  • it is possible to obtain a large Raman signal enhanced by a rod-like or block-like dry body structure formed on the periphery of the dry body it is possible to obtain a bowl-like or block-like dry body when observing the Raman spectrum. It is sufficient to obtain the Raman signal only by the structure. For detection of trace substances, it may take time to identify the observation site, but with this proposal, rapid observation is possible.
  • FIG. 1 A schematic view showing the work flow of the method for detecting a trace substance according to the present invention
  • ⁇ 4 A schematic diagram of the detection method of trace substances using a known SERS substrate (which is liquid, so it is not called a substrate! /, And so on) designed to stabilize metal particle colloids for a long period of time
  • FIG. 6 Flow chart of the method for detecting a trace substance of the present invention
  • FIG. 7 A data example of Raman spectrum by the detection method of the present invention.
  • N is not A or D, but is a position showing a Raman spectrum observed by moving the portion linearly and irradiating the laser.
  • FIG. 8 (a) is an optical micrograph of a bowl-like structure D formed on the peripheral portion R of S. (b) is an optical micrograph of a massive structure A formed on the periphery R of S.
  • a metal surface with a plasmon mirror effect for dropping a mixture of an analyte and metal fine particles A description of a solid plane having a hydrophilic / hydrophobic interface with a mixture of hydrophilic and hydrophobic surfaces.
  • the mixture in the step of dropping the mixture of the analyte and the metal fine particle colloid onto the solid plane, the mixture is dropped while rotating the mixture around an axis substantially perpendicular to the solid plane, or It is preferable to drop while rotating the solid plane about an axis substantially perpendicular to the solid plane. In this way, since the mixture is equalized in the rotational direction, a rod-like or block-like dry matter structure is uniformly formed at the peripheral edge, enabling quick observation without requiring time for identifying the observation site. It becomes.
  • FIG. 9 is an illustration of rotating the mixture about an axis generally perpendicular to the solid plane, or rotating the solid plane about an axis generally perpendicular to the solid plane.
  • the convex portion having a height of 1 ⁇ m or less or the concave portion having a height of 1 ⁇ m or less is provided on the solid surface on which the mixture of the analyte and the metal fine particle colloid is dropped.
  • a mixture of 1 to 10 microliters of analyte and 1 to 10 microliters of metal particulate colloid is dropped onto a solid surface, and a massive dry substance structure is formed in the vicinity of such irregularities. As a result, the observation site can be identified quickly without the need for time.
  • FIG. 10 is an explanatory view of a convex portion having a height of 1 ⁇ m or less and a concave portion having a height of 1 ⁇ m or less, which are disposed on a solid surface on which a mixture of an analyte and metal fine particles is dropped.
  • the mixture of the analyte and the metal fine particle colloid is dropped onto the solid surface of the hydrophilic / hydrophobic boundary portion having a hydrophilic / hydrophobic boundary portion where the hydrophilic surface and the hydrophobic surface are mixed.
  • FIG. 11 is an explanatory view of a solid plane on which a mixture of an analyte and metal fine particles is dropped and in which a hydrophilic surface and a hydrophobic surface are mixed and which has a hydrophilic / hydrophobic boundary portion.
  • the dried body according to the present invention is a dried body for detecting the analyte discrimination obtained by mixing and drying the analyte and the metal fine particle colloid, and 1 to 1: LO microliter analyte and 1 to 1 1 o
  • a mixture of micro metal particle colloids is dropped on a solid surface and dried at the periphery with a rod-like shape with a width of 1 to 10 ⁇ m or a massive structure with a major diameter of 1 to 10 ⁇ m.
  • the method of discriminating and detecting trace analytes by surface-enhanced Raman scattered light obtained by irradiating laser light to the site where the analytes and metal fine particles are close to each other. Is effective.
  • An apparatus for realizing the method of the present invention is characterized in that the mixing means for mixing the analyte and the metal microparticle colloid prepared in advance, and the mixture obtained by the mixing means into a solid plane, Means for blowing by pressure due to volume change of the piezoelectric element whose volume changes, drying means for drying the mixture sprayed by the means for blowing on a solid surface, and peripheral portions of a dried body of the mixture obtained by the drying means If it has a means to irradiate the laser beam and observe the Raman scattering light ,.
  • the apparatus according to the present invention is preferably a drying means for drying the mixture on a solid plane.
  • the apparatus according to the present invention is preferably provided with a pressure reducing means for lowering the solid plane below atmospheric pressure. (Claim 12), a dropping means for dropping the mixture of the analyte and the fine metal particle colloid onto the solid plane, a rotating means for rotating the mixture about an axis approximately perpendicular to the solid plane, or a solid plane approximately perpendicular to the solid plane It is preferable to combine the rotation means that rotates around the same axis.
  • the solid surface may be a metal surface having the brassmon mirror effect (see FIGS. 12 to 15).

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  • Analytical Chemistry (AREA)
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  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
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  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

Cette invention concerne un procédé et un dispositif permettant de détecter une substance à l'état de trace par spectrométrie laser de l'effet Raman exalté de surface, qui garantit une observation rapide et une mise en oeuvre remarquablement pratique. Ce procédé de détection d'une substance à l'état de trace par effet Raman exalté de surface englobe les opérations suivantes : mélange d'un analyte avec un colloïde de fines particules métalliques préconstitué ; stillation du mélange de l'analyste et dudit colloïde sur un plan solide; séchage du mélange sur le plan solide ; et irradiation de la partie frange du corps sec du mélange sur le plan solide par lumière laser et observation de la diffusion Raman de la lumière. Plus particulièrement, on irradie par une lumière laser une structure de corps sec présentant la forme d'un arbre ou d'une bosse formée sur la partie frange du corps sec et l'on observe la diffusion Raman de la lumière. Le plan solide peut être un plan métallique à effet miroir à plasmon. .
PCT/JP2006/323313 2005-11-22 2006-11-22 Procede et dispositif de detection d'une substance a l'etat de trace par effet raman exalte de surface Ceased WO2007060988A1 (fr)

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JP2005336513 2005-11-22
JP2005-336513 2005-11-22
JP2006022621 2006-01-31
JP2006-022621 2006-08-25

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010230352A (ja) * 2009-03-26 2010-10-14 Nidek Co Ltd 試験片，該試験片の製造方法、及び試験片を用いた測定法
WO2020026791A1 (fr) * 2018-07-31 2020-02-06 浜松ホトニクス株式会社 Procédé d'analyse d'analyte
CN110763670A (zh) * 2019-11-11 2020-02-07 山东师范大学 自分离多相混合液的拉曼增强活性衬底及制备方法与应用
CN112285087A (zh) * 2020-09-18 2021-01-29 山东师范大学 基于Au-Se界面的超灵敏高保真SERS传感器的制备及在生物小分子定量检测的应用
WO2021153253A1 (fr) * 2020-01-27 2021-08-05 浜松ホトニクス株式会社 Procédé d'analyse de cellule
JP2024534117A (ja) * 2021-08-24 2024-09-18 ホリバ インスツルメンツ インコーポレイテッド 流体試料中の汚染物質を検出するシステム及び方法

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH10160737A (ja) * 1996-12-03 1998-06-19 Dainippon Printing Co Ltd 光学的分析装置用測定チップ及びその製造方法
JPH1161209A (ja) * 1997-08-15 1999-03-05 Kdk Corp 貴金属微粒子の分散体及びその製造方法、並びに分散体を利用した構造体デバイス及びその製造方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH10160737A (ja) * 1996-12-03 1998-06-19 Dainippon Printing Co Ltd 光学的分析装置用測定チップ及びその製造方法
JPH1161209A (ja) * 1997-08-15 1999-03-05 Kdk Corp 貴金属微粒子の分散体及びその製造方法、並びに分散体を利用した構造体デバイス及びその製造方法

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010230352A (ja) * 2009-03-26 2010-10-14 Nidek Co Ltd 試験片，該試験片の製造方法、及び試験片を用いた測定法
CN112384788A (zh) * 2018-07-31 2021-02-19 浜松光子学株式会社 被检测物分析方法
WO2020026791A1 (fr) * 2018-07-31 2020-02-06 浜松ホトニクス株式会社 Procédé d'analyse d'analyte
JP2020020642A (ja) * 2018-07-31 2020-02-06 浜松ホトニクス株式会社 被検体分析方法
JP7190277B2 (ja) 2018-07-31 2022-12-15 浜松ホトニクス株式会社 被検体分析方法
CN110763670A (zh) * 2019-11-11 2020-02-07 山东师范大学 自分离多相混合液的拉曼增强活性衬底及制备方法与应用
WO2021153253A1 (fr) * 2020-01-27 2021-08-05 浜松ホトニクス株式会社 Procédé d'analyse de cellule
JP2021117106A (ja) * 2020-01-27 2021-08-10 浜松ホトニクス株式会社 細胞分析方法
CN115004014A (zh) * 2020-01-27 2022-09-02 浜松光子学株式会社 细胞分析方法
JP7344140B2 (ja) 2020-01-27 2023-09-13 浜松ホトニクス株式会社 細胞分析方法
CN112285087A (zh) * 2020-09-18 2021-01-29 山东师范大学 基于Au-Se界面的超灵敏高保真SERS传感器的制备及在生物小分子定量检测的应用
JP2024534117A (ja) * 2021-08-24 2024-09-18 ホリバ インスツルメンツ インコーポレイテッド 流体試料中の汚染物質を検出するシステム及び方法
JP7652983B2 (ja) 2021-08-24 2025-03-27 ホリバ インスツルメンツ インコーポレイテッド 流体試料中の汚染物質を検出するシステム及び方法

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