WO2007108360A1

WO2007108360A1 - Method for detecting substance by surface-enhanced raman scattering using small-diameter blind pipe, substance detecting device, device for detecting plurality of substances, and blind pipe

Info

Publication number: WO2007108360A1
Application number: PCT/JP2007/054897
Authority: WO
Inventors: Tomohiro Marui; Mayuko Yasuda
Original assignee: Intellectual Property Bank Corp.
Priority date: 2006-03-17
Filing date: 2007-03-13
Publication date: 2007-09-27
Also published as: JPWO2007108360A1

Abstract

This invention provides a method and apparatus for detecting a very small amount of a substance by surface-enhanced Raman scattering (SERS), which can realize excellent practicality and rapid observation. The method for detecting a very small amount of a substance by surface-enhanced Raman scattering comprises the step of mixing an analyte and a previously provided metal fine particle colloid to prepare a mixture for placing in a blind pipe in which a metal mass is provided in its blind part, the step of pouring the mixture of the analyte with the metal fine particle colloid into the blind pipe, and the step of applying a laser beam to a part around a solid-liquid interface where the mixture in the blind pipe is in contact with the blind part metal mass to observe Raman scattered light. The apparatus can realize the method.

Description

Specification

Substance detection method, substance detection apparatus, multi-substance detection device, blind tube using surface enhanced Raman scattering using small diameter blind tube

Technical field

[0001] The present invention relates to a method and an apparatus for detecting a trace substance by surface enhanced Raman scattering (SERS). In the present invention, surface-enhanced Raman scattering light obtained by irradiating a laser beam to a site where analytes, metal particles, and metal colloids are mixed together using a colloid in which metal particles or metal film and metal fine particles are suspended. In particular, the present invention uses a thin blind tube also called a Pasteur tube.

Background art

[0002] It is well known to detect analytes by Raman analysis by observing surface-enhanced Raman scattering (SERS) due to the plasmon effect of a group of noble metal nanoparticles such as gold and silver. It is. Even in the absence of nanoparticles, the Raman signal is enhanced by the plasmon effect and can be discriminated even with a very small amount of material with a weak Raman signal. The mechanism of SERS plasmon effect expression has not been fully elucidated, but research and development is being conducted for the purpose of application to harmful substance sensors. Here, the discrimination detection target substance is also called an analyte.

[0003] A method of obtaining a SERS-active solid substrate by dropping a SERS base material containing noble metal nanoparticles such as gold and silver and drying it is known. For example, Patent Document 1 discloses a simple method for preparing a substrate exhibiting high SERS activity by dispersing a SER S substrate in a liquid and fixing it on a solid surface.

[0004] A technique for obtaining strong SERS activity by self-assembling a group of nanoparticles has been studied. For example, a method for producing a SERS substrate in which metal fine particles are aligned by picking up a group of metal fine particles that are self-assembled at the interface in accordance with a manufacturing method of an LB film (Langmuir-Blodgett film) is also known (see Patent Document 3). Patent Document 2 is also a method of forming a SERS substrate in which particles are periodically aligned and deposited by repeatedly pulling up a flat plate immersed in a solution containing metal fine particles at a predetermined speed. [0005] Besides the nanoparticle alignment technique using LB films, many SERS substrate manufacturing methods have been proposed. For example, there is a method for producing a SERS substrate using a surface having affinity with water (hydrophilicity and water repellency) (see Patent Document 5).

[0006] A colloid in which precious metal nanoparticles suspended in a solid-state SERS active substrate as described above is suspended as a SERS substrate (not called a substrate because it is liquid), and this is a liquid phase mixture of analyte. It is also known to discriminate and detect analytes by observing surface-enhanced Raman scattering (see Patent Document 4).

[0007] A colloidal SERS substrate is easier to produce than a solid-phase SERS substrate, but has the disadvantage that the SERS activity disappears in the state of separation and precipitation and in the state of separation and precipitation. For this reason, methods for stabilizing a SERS substrate for a long period of time with a clay-like substance such as smectite (see Patent Document 4) have been studied.

[0008] It is also known to use capillary action (analysis) for analysis of a very small amount of analyte (see Patent Document 6). It is also known to detect and detect trace amounts of substances by mixing analyte and colloidal SERS substrate in a capillary phenomenon and analyzing the Raman signal of SERS.

[0009] Similarly, it is also possible to detect and detect trace amounts of substances by analyzing and mixing the SERS Raman signal by fluidly mixing microliter-level analytes and colloidal SERS substrates in a known microphone port channel. Known (see Patent Document 7 and Non-Patent Document 3).

[0010] On the other hand, Patent Document 8 discloses a trace substance detection method and apparatus using the "Blasmon mirror effect". Here, the plasmon mirror effect means that a group of nanoparticles is self-assembled (self-aligned) in the vicinity of a relatively clean metal surface that induces surface plasmon polaritons with the same applied light energy, and obtains stronger and SERS activity. That is. The conditions and mechanism for obtaining the plasmon mirror effect are not clear, but it is considered important to self-assemble the nanoparticle group in the vicinity of a relatively clean metal surface (Patent Document 8). reference).

[0011] On the other hand, the nanoparticles are self-assembled (self-aligning IJ) on the metal surface, and further metal deposition is performed on the upper surface, or the nanoparticles are stacked in two steps on the metal surface, thereby sandwiching the sandwich. “Sandwich Substrates” or “Sandwich architecture” is also known. Power, In some sandwich configurations, SERS activity stronger than a single layer may be obtained (see Patent Document 9, Non-Patent Document 4, and Non-Patent Document 5).

Patent Document 1: Japanese Unexamined Patent Application Publication No. 2005-077362 “Method for producing surface-enhanced Raman scattering active substrate” (Keio University)

Patent Document 2: Japanese Patent Application Laid-Open No. 2004-170334 “Raman scattering measurement sensor and manufacturing method thereof” (Japan Science and Technology Agency, etc.)

Patent Document 3: Japanese Laid-Open Patent Publication No. 2005-233637 “Raman Spectroscopic Analysis with Gold Nanorod Thin Film” (Japan Science and Technology Agency)

Patent Document 4: Japanese Patent Application Laid-Open Publication No. 2004-205435 “Analytical method of binding analysis target substance that does not require labeling dye and analysis kit used therefor” (Fukuo Takao)

Patent Document 5: Japanese Laid-Open Patent Publication No. 2005-219184 “Metal Nano-Triangular Column Structure for Single-Molecular Raman Spectroscopy, Array Substrate Formation Method and Single-Molecule Analysis Method Using the Same” (AIST) Patent Document 6: Patent No. 3462339 No. "Capillary electrophoresis detector" (Nippon Telegraph and Telephone Corporation)

Patent Document 7: Special Table 2005-507500 Publication “Detection of microfluids by surface-enhanced resonance Raman scattering” (University of Strathclyde et al.)

Patent Document 8: International Publication WO2005 114298 Publication `` OPTICAL SENSOR WITH LAYERED P LASMON STRUCTURE FOR ENHANCED DETECTION OF CHEMICAL GROUPS BY SERSJ VP HODLINGS'LLC

Patent Document 9: US Patent US6149868 `` Surface enhanced raman scattering from meta

1 nanoparticle-analyte-noble metal substrate sandwiches ”(Michael J. Natan) Non-patent document 1: Kotaro Kajikawa (Tokyo Tech), Yuta Mitsui“ Bio-sensing using localized plasmon resonance ”Applied Physics, No. 72, No. 12, p.1541-1544 (2003)

Non-Patent Document 2: Takayuki Okamoto (RIKEN) “Investigative Research on Metal Nanoparticle Interactions and Biosensors” 2002 Grant-in-Aid for Scientific Research (C) “Localization of surface plasmons and their applications Investigative research ”report on research results Individual report

Non-Patent Document 3: Keir R, Igata E, Arundell M, Smith WE, Graham D, McHugh C, Coop er JM. "SERRS. In situ substrate formation and improved detection using microfluidic sJ Anal Chem. 2002 Apr 1; 74 (7): 1503— 8 ·

Non-Patent Document 4: Jacquitta K. Daniels and George Chumanov (Clemson University) “Na noparticle-Mirror Sandwich Substrates for Surface-Enhanced Raman Scattering” _J. Phys. Chem. B, 109 (38), 17936 -17942, 2005.

Non-Patent Document 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 nanoparticle shape dependencej Anal Chem. 2005 May 15 ; 77 (10): 3261--6.

Disclosure of the invention

Problems to be solved by the invention

[0013] An object of the present invention is to propose a method and apparatus that are more practical than conventional methods and apparatuses for detecting trace substances using a SERS substrate or SERS substrate. In other words, we propose a method and apparatus that uses a colloidal SERS substrate but does not cause separation and precipitation of the substrate, without using a substrate in which metal fine particles are arranged through complicated processes. An object of the present invention is to propose an effective method and apparatus using both metal particles (or metal films) and metal colloids. Means for solving the problem

[0014] The method of the present invention (see claim 1, FIG. 1) is a surface-enhanced Raman scattering light obtained by irradiating laser light to a site where analyte, metal particles, and metal colloid are close to each other. A method for discriminating and detecting an analyte, in which the analyte and the metal fine particle colloid prepared in advance are provided in a blind tube in which a metal lump is disposed in the blind part or in a blind tube in which a metal film is disposed on the inner surface of the blind part And a step of injecting a mixture of the analyte and the metal fine particle colloid into the blind tube, and in the vicinity of the solid-liquid interface where the mixture of the injected blind tube is in contact with the blind metal mass. This is a method for detecting trace substances by surface-enhanced Raman scattering, which has a step of observing Raman scattered light by irradiating laser light.

[0015] Although there is no limitation on the inner diameter of the blind tube of the present invention, it is preferable that the analyte has a small amount of several μlitter, so that the inner diameter is preferably about 1 millimeter. If the mixture is sucked by capillary action (cylinder) and the opening is closed after suction, the inner diameter may be on the order of 100 micrometers. [0016] In the case of not using the fly, in order to inject the mixture of the analyte and the metal fine particle colloid into the blind portion, mechanical vibration including sound wave vibration may be applied from the outside in the vicinity of the blind portion. In other words, after the step of injecting the mixture of the analyte and the metal fine particle colloid into the blind tube, the step of inducing the mixture of the analyte and the metal fine particle colloid to the blind portion by applying vibration to the blind tube. You just have to. 2 in Fig. 2 is a means to vibrate the blind tube (a means to induce a mixture of A and C to the blind part in the blind tube by vibration). Figure 3 shows the Raman scattered light observation process when the blind tube is laid sideways, Figure 4 shows the Raman scattered light observation process when the blind tube is upright, and Figs. 5 and 6 show the observation example and figure. 7 is a photograph of the cecum and blind part used in the examples.

[0017] In the method of the present invention (Claim 3), before the step of mixing the analyte and the metal fine particle colloid prepared in advance, the position in the blind part of the metal mass or the inner surface of the blind part of the metal film It is preferable to have a step of measuring the position and arranging a marker at the position of the blind tube based on the measured position so that the measurement position information can be read by observation of the blind tube. Alternatively, it is preferable to have a step of arranging the storage medium storing the measurement position information in the blind tube so that the measurement position information can be read by observation of the blind tube. These aspects will be described later in the description of the device according to the present method.

The invention's effect

[0018] The present invention is more practical than conventional SERS substrates and methods and apparatuses for detecting trace substances using SERS substrates. This is because the detection can be carried out only with a blind tube such as glass and a pre-fabricated metal microparticle colloid, which is low in cost. A substrate in which metal fine particles are aligned through a complicated process is not used, but a colloidal SERS substrate is used, but the separation and precipitation of the substrate is not a problem. Even if a metal thin film is disposed on the inner surface of the blind tube, the cost is low. It can be said that this technique is suitable for automatic analysis of a large amount of various analytes.

Brief Description of Drawings

[0019] FIG. 1 is an explanatory view of a blind tube of the present invention. (A) is an example in which metal particles P, which are metal lumps, are arranged in the blind portion, (b) is an example in which the diameter of the metal particles is relatively small as in (a). (C) is an example in which the metal film S is arranged on the inner surface of the blind part, and (d) is the same as (c), and the arrangement part of the metal film S is arranged so as not to interfere with the Raman measurement from the side of the blind tube. An example of discontinuous dispersion.

FIG. 2 is a flow explanatory diagram of the detection method of the present invention. 2 by vibration to the blind in the blind tube An oscillating means having an ultrasonic vibrator or an eccentric rotor that applies mechanical vibration including sound waves to the blind tube by means of guiding the mixture.

Sono 3] Mixture This is an explanation of the process of observing Raman scattered light by irradiating laser light near the solid-liquid interface where it comes into contact with the blind metal block.

En 4] This is an explanation of the process of observing Raman scattered light as in Fig. 3, with the blind tube vertical. (A) is the part where the mixture AC with high focal position is not in contact with the blind metal block, and (b) is the part where the mixture AC is in contact with the blind metal block.

[Fig. 5] Data of an embodiment of the present invention, where Peakl is Raman data of a site where the analyte and metal colloid are close together, and Peak2 is a site where the analyte and metal colloid are close together. (Datal) Raman data, Raman data of the site (data2) where Peak2 analyte, metal particle, and metal colloid are close together. The analyte is adenosine.

[Fig.6] Spectral graph of Raman data (dataO) of the region where the analyte and metal are in close proximity, separately from the typical spectra (datal and data2) of Areal and Area2 in Fig. 5. The superiority of data2, the implementation result of this proposal, is clear.

7] Example of metal particles placed in the blind part of the blind tube

8] Explanation of device D with 1J parallel arrangement of multiple blind tubes 1 with metal lump arranged in the blind and / or multiple blind tubes 1 with metal film arranged inside the blind . leveU · -leveLn is the horizontal plane level including the position of the metal mass or metal film of the 1st · nth blind tube blind portion of the multiple blind tubes, dl · 'dn is leveU from the optical system reference part of the Raman measurement means, for example · The distance to 'leveLn' is set so that the optical focus of the optical system is all at the horizontal plane level including the position of the metal mass or metal film of the 1st · nth blind tube blind. To do.

9] An example of a blind tube in which the position of the metal lump (metal film) in the blind part is measured in advance and a storage medium is provided as a storage means for storing position information in the blind part of the metal lump (metal film). Position information Bl, B2, B3 for the optical marker M placed in the blind part of the blind canal as a reference for the measurement position is encoded by the encoding means 4 and printed by the printing means 5, for example, a bar code 9 (printed on the blind pipe) Printed as Bl, B2 and B3).

10] An example of a blind tube in which the position of the metal lump (metal film) in the blind portion is measured in advance, and a marker is disposed at the position of the blind tube based on the measured position, as shown in FIG. Where Position information Bl, B2, B3 is grasped from the marker placement position itself. Ml is the first optical marker placed in the blind canal after the actual measurement of the position of the metal lump or metal film, and M2 is the blind in the blind canal after the actual measurement of the position of the metal lump or metal film. The arranged second optical marker, ML, is a predetermined distance apart from the position of the metal block or metal film (the marker position is separated so as not to obstruct the Raman measurement).

[Fig.11] There is a margin for grinding 7 at the end of the blind tube, and the grinding margin 7 is ground according to the position of the metal mass of the blind part or the metal film on the inner surface of the blind part, and multiple blind pipes are ground to the reference surface An example of parallel arrangement with aligned surfaces. 7 is the margin of the bottom of the blind tube for leveling the metal lump or metal film, 8 is the bottom of the blind tube after being ground for leveling, G is the reference plane corresponding to the focal point of the laser beam irradiation , H is one of the focal planes of the actual laser beam irradiation, and is the highest level in the horizontal plane including the position of the metal block or metal film, and J is a certain level difference from the actual focal plane H of the reference plane G.

[FIG. 12] A cone-shaped reference shape 7A is provided at the end of the blind tube, and has a reference surface corresponding to the focal point of the laser beam irradiation, and a conical knife that fits the above-described cone-shaped shape on the reference surface. The mold is opened, and the depth of the cone-female mold is determined according to the position of the blind metal block or the metal film on the inner surface of the blind part. As a result, a plurality of blind tubes are arranged in parallel by fitting the cone-shaped reference shape 7A at the end of the blind tube into the conical female-type opening. 7A is a cone-shaped reference shape for leveling the metal mass or metal film position.

[FIG. 13] This is an example of a blind tube according to the present invention, in which (a) is a method of charging a metal material into a glass empty blind tube, (b) is a metal melting by heating the blind portion of a glass empty blind tube, c) shows the steps of forming the constricted part to prevent the metal from falling off by pressurizing and constricting the blind part.

Explanation of symbols

1 Blind tube with a metal block on the blind part or a blind tube with a metal film on the inner surface of the blind part

2 Means to vibrate the blind tube (Means to induce a mixture of A and C into the blind part of the blind tube by vibration)

Means to input 3 Bl, B2, B3 data and output to encoding means

4 Encoding means

5 Printing method 6 Trace substance detection chip with multiple blind tubes 1 arranged in parallel

7 Grind the bottom of the blind tube for leveling the metal mass or metal film position

7A metal mass or reference shape in the shape of a cone for leveling a metal film

8 Blind tube bottom after being ground for leveling

9 Code encoded Bl, B2, B3 printed on the blind tube

A Analyte

Mixture of AC analyte and colloidal metal

ACP AC is close to P (or metal film)

Areal Analyte and colloidal metal

Area2 Area where the analyte, metal particles, and metal colloids are close together

Bl M force, the distance to the closest surface part of P

B2 M force, distance to the farthest surface part of P

Distance from B3 M to F

C Metal colloid

D Device in which multiple blind tubes 1 with metal blocks and / or metal films arranged in the blind are arranged in parallel

dl For example, the distance from the optical system reference part of the Raman measurement means to leveLl

d2 For example, the distance from the optical system reference part of Raman measurement means to level_2

d3 For example, the distance from the optical system reference part of Raman measurement means to level_3

dn For example, the distance from the optical system reference part of Raman measurement means to level_4

dataO Raman data of the site where the analyte and metal are in close proximity

datal Raman data of the site where the analyte and metal colloid are close together

data2 Raman data of the site where the analyte, metal particles, and metal colloid are in close proximity

Site where FPs are close enough and SERS effect is remarkable

G Reference plane corresponding to laser beam irradiation focus

H One of the focal planes of actual laser light irradiation, the highest level in the horizontal plane including the position of the metal lump or metal film J A certain level difference from the reference plane G to the actual focal plane H

level.l Horizontal plane level including the position of the metal mass or metal film of the first cephalic blind region of multiple blind tubes

level_2 The horizontal plane including the position of the metal mass or metal film of the second blind canal of multiple blind tubes

level_3 Horizontal plane level including the location of the metal mass or metal film of the 3rd culm

level n Horizontal plane level including the position of the metal mass or metal film of the nth cephalic blind region of several culm

M Optical marker placed in the blind part of the blind tube as a reference for the measurement position

After the actual measurement of the position of the metal lump or metal film by Ml measurement,

1 Optical marker

M2 Second optical marker placed in the blind canal after the actual measurement of the position of the metal mass or metal film

ML Predetermined distance away from the position of the metal mass or metal film (the marker position is separated so as not to obstruct Raman measurement)

P metal particles

A characteristic peak of adenosine (analyte) in Peakl datal

Peak2 A characteristic peak of adenosine (analyte) in data2

PL1 The highest level in the horizontal plane including the position of the metal mass or metal film of the first blind canal

PL2 The highest level in the horizontal plane, including the location of the metal mass or metal film of the second blind canal

PL3 The highest level in the horizontal plane including the position of the metal mass or metal film of the third blind canal

PLU The highest level in the horizontal plane, including the position of the metal mass or metal film in the blind canal

The lowest level in the horizontal plane, including the position of the metal mass or film in the PLL blind tube

QL1 A level corresponding to the PL1 level. It corresponds to a female cone to be fitted.

QL2 Level corresponding to PL2 level Corresponds to the female cone to be fitted.

QL3 A level corresponding to the PL3 level. It corresponds to a female cone to be fitted.

S Metal film inside the blind part BEST MODE FOR CARRYING OUT THE INVENTION

[0021] The device according to the present invention (Claim 5) includes a blind tube in which a metal lump is disposed in the blind part, or a blind tube in which a metal film is disposed on the inner surface of the blind part, and laser light is applied to the blind tube. The means for observing the Raman scattered light by irradiation, and the blind tube into which the mixture of the analyte and the metal fine particle colloid is injected, the focus of the laser beam near the solid-liquid interface where the mixture and the metal lump or metal film are in contact with each other. And means for matching.

[0022] Further, the device according to the present invention (Claim 6) may be provided with means for applying vibration to the blind tube to guide the mixture of the analyte and the metal fine particle colloid to the blind part.

[0023] Further, the device according to the present invention (Claim 7) is provided in the position of the blind portion of the metal mass or the inner surface of the blind portion of the metal film before the step of mixing the analyte and the colloidal metal colloid prepared in advance. The position is measured in advance, and a blind tube in which a single force is arranged at the position of the blind tube based on the measurement position, and the detection device has a means for reading the marker position and also has a fixed position. The means for focusing the laser beam near the liquid interface discriminates and focuses the solid-liquid interface based on the marker position read by the reading means. (Refer to the brief explanation of Fig. 10 and Fig. 10.)

[0024] In addition, here, the marker is located most in the plane group perpendicular to the tube axis of the blind tube and including the blind metal mass or the metal film of the blind inner surface. The position included in the plane close to the open end of the cephalic tube (see PLU in Figure 4) and / or the position closest to the blind end of the cephalic canal and the position of the plane (see PLL in Figure 4) is preferred.

[0025] As another aspect (Claim 9), the storage means for storing the measurement position information and the information reading means for the storage means are combined, and the laser beam is focused near the solid-liquid interface. The means for determining and focusing the solid-liquid interface based on the position information on the blind portion of the metal lump read by the reading means or the position information on the inner surface of the blind portion of the metal film may be used. In particular (Claim 10), a storage medium as storage means for storing measurement position information in the blind tube is provided, and information reading means in the storage means is information reading means in the storage medium provided in the blind tube. Is preferred (see brief description of FIGS. 9 and 9).

[0026] Further, the present invention provides a multiple detection for detecting a minute amount of analyte latent in a plurality of sampling solutions. It may constitute a several substance detection device. Here, it is hoped that the position of the metal inside the plurality of blind tubes approximately coincides with the axial direction of the blind tubes so that the focusing of the laser light can be omitted in the vicinity of each solid-liquid interface. Masle.

[0027] That is, (Claim 11), a plurality of blind tubes with a metal lump disposed in the blind portion and a plurality of blind tubes with a metal film disposed on the inner surface of the Z or blind portion under the following parallel conditions: The parallel condition is as follows: the parallel condition is: in a plane perpendicular to one blind tube axis and in a group of planes including a blind metal mass or a metal film on the inner surface of the blind The surface closest to the open end of the blind canal or the surface closest to the blind canal is perpendicular to the axis of the other canal and is included in the group of surfaces including the blind metal mass or the metal film on the inside of the blind (See Figure 8).

[0028] Specifically, this is a multi-substance detection device having the following structure (refer to claims 12, FIG. 11 and a brief description of FIG. 11). In other words, 1) There must be an IJ margin at the end of the blind tubes before the parallel arrangement. 2) The multiple substance detection device has a reference surface corresponding to the focal point of the laser beam irradiation. 3) The grinding margin at the end of the blind pipe is ground according to the position of the metal block of the blind block or the metal film on the inner surface of the blind block. 4) A plurality of blind tubes are arranged in parallel with the ground surface ground to the reference surface.

[0029] Another specific embodiment (see the brief description of claim 13, FIG. 12 and FIG. 12) is as follows: 1) There is a conical oscillating reference shape at the end of the blind tube before the parallel arrangement. . 2) The multi-substance detection device has a reference plane corresponding to the focal point of the laser beam irradiation, and a conical knife type that fits the circular cone type is opened on the reference plane. 3) The depth of the conical knife type is determined in accordance with the position of the blind metal block or the metal film on the inner surface of the blind part. 4) A plurality of blind tubes are arranged in parallel by fitting the cone-shaped reference shape at the end of the blind tube into the conical female opening.

A preferred embodiment of the blind tube of the present invention is shown in FIG. 13 together with its production method. That is, (Claim 14) is a blind tube in which metal is disposed in the blind part, and after the step of introducing the metal material into the blind tube (FIG. 13 (a)), the metal in the blind part is melted. A blind tube in which the input metal material is melted and fixed to the inner surface of the blind tube blind part by the step of heating above the temperature (FIG. 13 (b)) is suitable.

[0031] (Claim 15) A blind tube in which a thin film in which the above-mentioned metal material is melt-extended is disposed on the inner surface of the blind tube blind portion by the same metal material charging step and heating step is also suitable. [0032] Furthermore, (Claim 16) is a blind tube in which metal is disposed in the blind portion, and after the step of disposing the metal material in the blind tube, the open end side of the blind tube metal disposed portion. Even if a part having an inner diameter smaller than the inner diameter of the blind tube metal placement part is formed on the open end side of the metal placement part by the process of constricting the pressure from the outside of the blind pipe (Fig. 13 (c)) Good. This small inner diameter portion is suitable because it is difficult for the metal material to separate from the blind portion without being melt-fixed or melt-extended by heating.

[0033] In the process of heating the metal in the blind portion above the melting temperature, if the metal has a relatively low melting temperature, such as gold, the blind tube is heated from outside (see FIG. (See Fig. 13)

[0034] Of course, if the blind tube member is made of heat resistant material (heat resistant glass), it is easy to heat, and only the metal in the blind tube blind part is locally heated by local heating means such as laser heating to avoid melting of the blind tube material. You can do it. Further, all or part of the metal may be a low melting point alloy having a low melting point, and only the alloy may be melted.

Claims

The scope of the claims

[1] A method for discriminating and detecting trace amounts of analytes using surface-enhanced Raman scattering light obtained by irradiating laser light to a site where analytes, metal particles, and metal colloids are close to each other. A process of mixing the analyte with a metal particulate colloid prepared in advance in a blind tube in which a lump is disposed or a metal film disposed on the inner surface of the blind portion, Injecting the mixture into the blind tube and observing Raman scattered light by irradiating a laser beam in the vicinity of the solid-liquid interface where the mixture of the injected blind tube is in contact with the blind metal mass A method for detecting trace substances by surface-enhanced Raman scattering.

[2] In the detection method according to claim 1, after the step of injecting the mixture of the analyte and the metal fine particle colloid into the blind tube, the mixture of the analyte and the metal fine particle colloid is blinded by applying vibration to the blind tube. A method for detecting a trace substance by surface-enhanced Raman scattering, further comprising a guiding step.

[3] In the detection method according to claim 1, before the step of mixing the analyte and the metal fine particle colloid prepared in advance, the position of the metal block in the blind part or the position of the metal film in the blind part inner surface is determined. A method of detecting a trace substance by surface-enhanced Raman scattering, further comprising a step of measuring and arranging a marker at a position of the blind tube based on the measurement position so that the measurement position information can be read by observation of the blind tube.

[4] In the detection method according to claim 1, before the step of mixing the analyte and the metal fine particle colloid prepared in advance, the position of the metal block in the blind part or the position of the metal film in the blind part inner surface is determined. A method of detecting a trace substance by surface-enhanced Raman scattering, further comprising a step of measuring and arranging a storage medium storing the measurement position information in a blind tube so that the measurement position information can be read by observation of the blind tube .

[5] A device for discriminating and detecting a small amount of analyte by the method according to claim 1, wherein the blind tube has a metal lump disposed in the blind part, or a blind tube in which a metal film is disposed on the inner surface of the blind part. The means for observing Raman scattered light by irradiating the blind tube with laser light, and the blind tube injected with a mixture of the analyte and the metal fine particle colloid, An apparatus for detecting a trace substance having means for focusing a laser beam in the vicinity of a solid-liquid interface in contact with.

[6] The detection device according to claim 5, wherein the analyte and the metal fine particle roller are vibrated by applying vibration to the blind tube. A device for detecting trace substances that also has a means for guiding a mixture of ids to the blind.

[7] Before the step of mixing the analyte and the metal fine particle colloid prepared in advance, the blind tube of the detection device according to claim 5 is positioned in the blind portion of the metal lump or the inner surface of the blind portion of the metal film. Is a blind tube in which a marker is arranged at the position of the blind tube based on the measured position,

6. The detection apparatus according to claim 5, wherein the means for reading the marker position is combined, and the means for focusing the laser beam in the vicinity of the solid-liquid interface is based on the marker position read by the reading means. Detecting device for trace substances that discriminate and focus.

[8] The position of the marker according to claim 7 is a plane that is perpendicular to the tube axis of the bleed tube, and is the most open end of the bleed tube in the group of surfaces including the metal mass of the blind part or the metal film of the inner surface of the blind part. Detecting device that corresponds to the position included in the surface close to and the position of the surface closest to Z or the blind end of the blind canal.

[9] In the blind tube of the detection device according to claim 5, before the step of mixing the analyte and the metal fine particle colloid prepared in advance, the position in the blind portion of the metal lump or the inner surface of the blind portion of the metal film The position at is intensively measured,

The detection device according to claim 5, wherein the storage unit that stores the measurement position information and the information reading unit of the storage unit are combined, and the unit that focuses the laser beam in the vicinity of the solid-liquid interface, An apparatus for detecting a trace substance that distinguishes and focuses a solid-liquid interface based on position information in a blind portion of a metal lump read by the reading means or position information on an inner surface of a blind portion of a metal film.

10. The storage device according to claim 9, wherein a storage medium that is a storage unit that stores measurement position information is disposed in the blind tube of the detection device, and the information reading unit of the storage unit is disposed in the blind tube. A trace substance detection device that is a medium information reading means.

[11] A multi-substance detection device for discriminatingly detecting trace analytes latent in a plurality of sampling solutions by the method according to claim 1, comprising a plurality of blind pipes having a metal lump disposed in a blind portion and A device in which multiple blind tubes with metal films on the inner surface of the blind part are placed in parallel under the following parallel conditions. The parallel condition is:

The plane that is perpendicular to the axis of the single blind tube and that is the surface closest to the open end of the blind tube or the surface closest to the blind end of the blind tube in the group of surfaces including the blind metal mass or the metal film on the inner surface of the blind Is included in the plane group that is perpendicular to the other tube axis and includes the blind metal mass or the metal film on the inner surface of the blind.

[12] A multiple substance detection device having the following configuration in order to realize the parallel condition of a plurality of blind tubes according to claim 11.

1) There must be a margin for grinding at the end of the blind tubes before parallel arrangement.

2) The multiple substance detection device has a reference surface corresponding to the laser beam irradiation focus.

3) Grinding margin at the end of the blind tube should be improved according to the position of the metal block of the blind portion or the metal film on the inner surface of the blind portion.

4) A plurality of blind tubes are arranged in parallel with the ground surface ground to the reference surface.

[13] A multiple substance detection device having the following configuration in order to realize the parallel condition of a plurality of blind tubes according to claim 11.

1) There should be a cone-shaped reference shape at the end of the blind tube before the parallel arrangement.

2) The multi-substance detection device has a reference plane corresponding to the focal point of the laser beam irradiation, and a conical knife type that fits with the conical male type is opened on the reference plane.

3) The depth of the conical knife type is determined according to the position of the blind metal block or the metal film on the inner surface of the blind part.

4) A plurality of blind tubes are arranged in parallel by fitting the cone-shaped reference shape at the end of the blind tube into the conical female opening.

[14] A blind tube in which a metal is disposed in the blind portion, and the step of heating the metal in the blind tube blind portion to a melting temperature or higher after the step of introducing a metal material into the blind tube is performed. A blind tube for detecting trace substances by surface-enhanced Raman scattering in which the injected metal material is melted and fixed on the inner surface of the blind part.

[15] A blind tube in which a metal is disposed in the blind part, and the step of heating the metal in the blind part to a temperature equal to or higher than the melting temperature after the step of introducing a metal material into the blind pipe, Trace material inspection by surface-enhanced Raman scattering in which a thin film in which the injected metal material is melt-extended is disposed on the inner surface of the blind part Intellectual blind tube.

A blind tube in which metal is disposed in the blind portion, and after the step of disposing the metal material in the blind tube, the step of pressurizing and constricting the open end side of the blind tube metal disposing portion from the outside of the blind tube A blind tube for detecting trace substances by surface-enhanced Raman scattering, wherein a portion having an inner diameter smaller than the inner diameter of the blind tube of the blind tube metal arrangement portion is formed on the open end side of the metal arrangement portion.