WO2019221298A1 - Élément d'amplification des ondes électromagnétiques, son procédé de production, procédé de détection utilisant l'élément d'amplification des ondes électromagnétiques et procédé de détermination d'une séquence d'acides aminés - Google Patents

Élément d'amplification des ondes électromagnétiques, son procédé de production, procédé de détection utilisant l'élément d'amplification des ondes électromagnétiques et procédé de détermination d'une séquence d'acides aminés Download PDF

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WO2019221298A1
WO2019221298A1 PCT/JP2019/019810 JP2019019810W WO2019221298A1 WO 2019221298 A1 WO2019221298 A1 WO 2019221298A1 JP 2019019810 W JP2019019810 W JP 2019019810W WO 2019221298 A1 WO2019221298 A1 WO 2019221298A1
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electromagnetic wave
metal
dielectric
enhancing element
metal part
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PCT/JP2019/019810
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English (en)
Japanese (ja)
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粟屋信義
西富雄
梶田浩志
田中覚
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Ｓｃｉｖａｘ株式会社
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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

Definitions

  • the present invention relates to an electromagnetic wave enhancing element, a production method thereof, and an amino acid sequence determination method using the electromagnetic wave enhancing element.
  • Raman spectroscopy is a method for determining the molecular structure and crystal structure of a substance by examining the properties of Raman scattered light, which is light having a wavelength different from that of incident light.
  • the intensity of Raman scattered light is extremely weak, about 10 to the sixth power of the intensity of Rayleigh scattered light. Therefore, it is necessary to use a high-intensity light source such as a laser practically.
  • SERS surface-enhanced Raman scattering
  • SERS is a phenomenon in which Raman scattering of molecules adsorbed on a metal surface having a nano-order fine structure is remarkably enhanced.
  • the mechanism of this enhancement includes the enhancement of vibration (chemical enhancement) by charge transfer between metal and molecule, and the enhancement of the electric field formed when localized surface plasmons are excited by incident light (physical enhancement). ).
  • Patent Document 1 As a method for forming a nanogap structure in a self-organized manner by surface treatment, a method using a porous silicon structure (for example, Patent Document 1), a method for forming a nanostructure by embossing (for example, Patent Document 2) ), A method of forming a nanostructure by surface roughening by boehmite treatment (for example, Patent Document 3).
  • a method for forming a nanostructure in bulk a method for forming a nanocomposite (for example, Patent Document 4), a method for forming a metal nanosponge (for example, Patent Document 5), and a method for aggregating fullerenes (for example, Patent Documents) 6).
  • Patent Document 7 Also proposed is a technique in which metal particles are dispersed and fixed on the surface of a substrate to form nano-sized intervals in a self-organized manner (for example, Patent Document 7).
  • a metal film is deposited on the stepped pattern by a film forming method with insufficient step coverage such as vapor deposition or sputtering to form a nano-sized gap in a self-aligned manner in the stepped portion.
  • a film forming method with insufficient step coverage such as vapor deposition or sputtering to form a nano-sized gap in a self-aligned manner in the stepped portion.
  • Patent Document 8 There is a method of forming (for example, Patent Document 8).
  • Patent Document 9 As a structure using plasmon resonance in combination, a structure in which metal particles are periodically arranged via a metal mirror and an insulating film has been proposed (for example, Patent Document 9).
  • JP 2014-178327 A Special table 2009-501904 JP2014-202650 JP2015-68736 Special table 2011-533677 JP 2014-159364 A JP-A-2005-233637 JP2015-14547 Special table 2007-538264
  • the method of surface treatment, the method of forming nanostructures in bulk, and the method of spraying metal nanoparticles are not methods that can be processed uniformly and precisely, and thus the size of the nano-sized gap cannot be controlled. Therefore, there is a problem that the reproducibility of the SERS enhancement cannot be obtained.
  • the distance between the metal mirror and the metal particles can be controlled by controlling the thickness of the insulating film.
  • the shape of each uneven structure is not optimized, and the electric field enhancement effect is not sufficient.
  • an object of the present invention is to provide an electromagnetic wave enhancing element that brings about an electric field enhancement as compared with the conventional one and enables quantitative evaluation.
  • the electromagnetic wave enhancing element includes a dielectric concavo-convex structure formed by extending a dielectric portion in which a dielectric concavo-convex structure is formed and a concave portion whose width decreases toward the bottom side in one direction on the dielectric concavo-convex structure. And a metal part.
  • the concave portion when the angle formed by the tangents at the same height point of adjacent convex portions forming the concave portion is ⁇ , the concave portion preferably has a minimum value of ⁇ of 20 degrees or less.
  • the metal part is preferably A> B, where A is the vertical thickness of the uppermost point of the convex part of the metal concavo-convex structure, and B is the vertical thickness of the lowermost point. .9A> B is preferred.
  • the metal part is A> C, where A is the vertical thickness of the uppermost point of the convex part of the metal concave-convex structure and C is half the width of the lowest point of the concave part of the dielectric concave-convex structure. It is more preferable that A> 2C.
  • the metal part includes a metal part main body for defining the metal uneven structure, and a metal layer formed on a surface of the metal part main body and made of a metal having a higher electric field strength than the metal part main body. May be.
  • the pitch of the metal concavo-convex structure is preferably 200 nm or less.
  • the convex surface has a curved shape that swells toward the concave side.
  • the dielectric concavo-convex structure has a plurality of types of shapes.
  • the dielectric uneven structure may be composed of a plurality of regions, and the dielectric uneven structure may have a different shape for each region.
  • the metal concavo-convex structure is bonded with a fixing substance for fixing a specific substance.
  • the immobilizing substance can immobilize amino acids.
  • hydrophilic layer having hydrophilicity may be provided on the surface of the metal part.
  • an intermediate layer for improving the adhesion between the dielectric part and the metal part may be provided between the dielectric part and the metal part.
  • the method for manufacturing an electromagnetic wave enhancing element according to the present invention includes a dielectric part forming step of forming a dielectric uneven structure on the dielectric part, and a concave part that decreases in width toward the bottom side on the dielectric uneven structure.
  • the concave part is set so that the minimum value of ⁇ is 20 degrees or less. It is preferable to form the film.
  • the metal part is formed such that A> B, where A is the vertical thickness of the uppermost point of the convex part of the metal concavo-convex structure and B is the vertical thickness of the lowermost point. Is more preferable, and it is more preferable to form the metal portion so that 0.9A> B.
  • A> C where A is the thickness in the vertical direction of the top point of the convex portion of the metal concavo-convex structure, and C is the half width of the bottom portion of the concave portion of the dielectric concavo-convex structure.
  • a metal part main body for defining the metal concavo-convex structure is formed, and a metal layer made of a metal having a higher electric field strength than the metal part main body is formed on the surface of the metal part main body. It may be.
  • the dielectric part forming step is such that the pitch of the dielectric uneven structure is formed to be 200 nm or less.
  • the metal part forming step it is preferable to control the film thickness by using a film forming technique that increases the growth rate in the vertical direction of the metal part.
  • a film forming technique that increases the growth rate in the vertical direction of the metal part.
  • the metal part formed by the sputtering may be further subjected to electroless plating to adjust the shape of the metal part.
  • an intermediate layer forming step of forming an intermediate layer for improving the adhesion between the dielectric portion and the metal portion on the dielectric portion may be provided.
  • the metal part may have a fixed substance binding step of binding a fixed substance capable of fixing a specific substance.
  • It may have a hydrophilic layer forming step of forming a hydrophilic layer having hydrophilicity on the surface of the metal part.
  • the amino acid sequence determination method of the present invention includes a sequential decomposition step of sequentially decomposing amino acids from the N-terminus or C-terminus of a peptide or protein, a fractionation step of fractionating amino acids released by the sequential decomposition step, And an analysis step of analyzing the amino acid obtained by the fractionation step using the electromagnetic wave enhancing element of the present invention.
  • the sequential decomposition step may be performed using a protease.
  • the sequential decomposition step may be performed using a column on which the protease, the peptide, or the protein is immobilized.
  • the electromagnetic wave enhancing element of the present invention can bring about a large electric field enhancement by optimizing the shape of the metal concavo-convex structure.
  • 6 is a graph showing the maximum Raman intensity of Samples 1 to 3 and Comparative Samples 1 and 2. 6 is a graph showing the relationship between the size of the gap between samples 4 to 6 and the Raman intensity. It is sectional drawing which shows an electromagnetic wave enhancing element. It is sectional drawing which shows another electromagnetic wave enhancing element of this invention. It is sectional drawing which shows another electromagnetic wave enhancing element of this invention. It is sectional drawing which shows the electromagnetic wave enhancing element of the present invention. It is sectional drawing which shows another electromagnetic wave enhancing element of this invention.
  • the electromagnetic wave enhancing element of the present invention is a metal having a shape in which a dielectric portion 1 in which a dielectric concavo-convex structure 11 is formed and a concave portion 22 whose width decreases toward the bottom side is extended in one direction.
  • the recess 22 should have a minimum gap width of 25 nm or less, preferably 20 nm or less, between adjacent protrusions forming the recess 22. As shown in FIG.
  • the recess 22 is preferably in contact with adjacent projections 24A and 24B forming the recess 22, but as long as the electric field can be sufficiently enhanced, FIG. As shown in (b), there may be a gap. Further, in FIG. 1A, there is no gap between the dielectric part 1 and the metal part 2, but if the electric field can be sufficiently enhanced, the dielectric part 1 and the metal part 2 can be seen as shown in FIG. There may be a gap 29 between them.
  • the metal part 2 has a metal concavo-convex structure 21 constituting a concave part 22 for concentrating the electric field.
  • the metal concavo-convex structure 21 is formed in a shape obtained by extending the recess 22 in one direction, that is, linearly in a plan view.
  • an electromagnetic wave having a predetermined wavelength ⁇ is incident on the metal concavo-convex structure 21, a strong electric field is concentrated in a very narrow region of the concave portion 22 of the metal concavo-convex structure 21.
  • the SERS signal can be taken out using the portion showing the maximum value of the electric field enhancement intensity.
  • the angle formed between the tangents 25A and 25B at the same height point (the lowest point 23 of the concave portion 22 in the figure) of the adjacent convex portions 24A and 24B forming the concave portion 22 is defined.
  • the metal concavo-convex structure 21 having a large maximum value Ex of the electric field enhancement strength tends to be large when the minimum value of the angle ⁇ is 20 degrees or less.
  • the metal concavo-convex structure 21 having a large maximum value Ex of the electric field enhancement strength tends to increase when 0 ° ⁇ ⁇ 15 °, and tends to increase when 7 ° ⁇ ⁇ 13 °. .
  • the point of contact with the convex portion 24 of the tangent where ⁇ is the minimum value is half of the height of the convex portion 24 (the height of the highest point 26 of the convex portion 24 with respect to the lowest point 23 of the convex portion 24). It is better to be on the lower side, preferably lower than a quarter of the height of the convex portion 24, more preferably at the lowest point 23. Therefore, it is preferable that the shape of the convex portion 24 is a curved shape in which the surface of the convex portion 24 swells toward the concave portion 22 as shown in FIG.
  • the angle ⁇ may be measured by taking a cross-sectional photograph perpendicular to the extending direction of the metal concavo-convex structure 21 and analyzing the image.
  • the metal concavo-convex structure 21 having a large thickness tends to be large.
  • the metal concavo-convex structure 21 having a large maximum value Ex of the electric field enhancement strength tends to increase when 0.9A> B, and further tends to increase when 0.6A> B.
  • the electric field enhancement when the vertical thickness of the uppermost point 26 of the convex portion 24 of the metal concavo-convex structure 21 is A and the half width of the bottom of the concave portion 12 of the dielectric concavo-convex structure 11 is C, the electric field enhancement When the film thickness A is larger than C, the metal uneven structure 21 having a large maximum value Ex tends to increase. In particular, the metal concavo-convex structure 21 having a large maximum value Ex of the electric field enhancement strength tends to increase when A> 2C, and further tends to increase when A> 3.5C. In addition, as shown in FIG.
  • a gap is formed in the concave portion 22 of the metal concavo-convex structure 21, or a gap is formed between the dielectric portion 1 and the metal portion 2 as shown in FIG. Even in the case of 29, it is better that A> C, preferably A> 2C, and more preferably A> 3.5C.
  • the pitch P of the convex portions 24 of the metal concavo-convex structure 21 may be 1000 nm or less, preferably 200 nm or less, and more preferably 120 nm or less.
  • the material of the metal part 2 may be any material as long as it reflects electromagnetic waves.
  • metals such as gold, silver, copper, chromium, aluminum, platinum, and tungsten can be used. Moreover, these combinations may be sufficient.
  • the metal part 2 does not need to consist of a single metal, and a metal part body for defining a metal uneven structure and a metal layer made of different metals formed on the surface of the metal part body. It may be configured. For example, depending on the material of the metal part 2, there are some that have a large electric field enhancement but are difficult to form the metal relief structure 21. In this case, as shown in FIG. 22, the metal part 2 is formed on the surface of the metal part main body 2A for defining the metal uneven structure 21 and the metal part main body 2A, and has a higher electric field strength than the metal part main body 2A. And a metal layer 2B made of In this case, the metal layer is preferably 1 nm or more.
  • the dielectric portion 1 has a dielectric concavo-convex structure 11 for controlling the shape and pitch of the metal concavo-convex structure 21.
  • the material of the dielectric part 1 may be any dielectric material that can form the dielectric concavo-convex structure 11.
  • a resin such as an acrylic resin or a cyclic olefin resin, silicon (Si), An inorganic compound such as silicon dioxide (SiO 2 ) can be used.
  • a photocurable resin or a thermoplastic resin suitable for the imprint method may be used.
  • the shape of the dielectric concavo-convex structure 11 may be any shape as long as the shape of the metal concavo-convex structure 21 can be controlled so as to have a concave portion 22 whose width decreases toward at least the bottom side.
  • the shape of the dielectric concavo-convex structure 11 is preferably one that can control the shape of the metal concavo-convex structure 21 so that the minimum value of the angle ⁇ described above is 20 degrees or less.
  • the surface of the convex portion 14 of the dielectric concavo-convex structure 11 can be a curved shape swelled toward the concave portion 22 side.
  • the shape of the metal concavo-convex structure 21 can be controlled, a cross section having a trapezoidal shape, a rectangular shape such as a rectangle or a square, a triangular shape, or the like may be used.
  • the pitch of the dielectric concavo-convex structure 11 may be the same as the pitch P of the metal concavo-convex structure 21.
  • the other shapes of the dielectric concavo-convex structure 11 may be any shape as long as the metal concavo-convex structure 21 can be controlled so that the electromagnetic field can be enhanced, for example, the width and aspect ratio of the convex portion 14.
  • the width of the convex portion 14 of the dielectric concavo-convex structure 11 may be 30 to 60% of the pitch.
  • the aspect ratio of the convex portion 14 of the dielectric concavo-convex structure 11 is preferably 1 or more.
  • the enhancement of electromagnetic waves is proportional to the fourth power of the electric field, the electric field varies greatly depending on a slight difference in the shape of the metal concavo-convex structure 21.
  • the metal concavo-convex structure 21 has a sufficient number of recesses 22 so as to include a shape capable of enhancing electromagnetic waves.
  • the dielectric concavo-convex structure 11 formed on the electromagnetic wave enhancing element may have a plurality of types of shapes as shown in FIG.
  • the metal concavo-convex structure 21 to be formed also has a plurality of types of shapes, and the metal concavo-convex structure 21 that can enhance electromagnetic waves is reliably formed. can do.
  • the enhancement of electromagnetic waves is proportional to the fourth power of the electric field, the electromagnetic wave can be sufficiently enhanced if there is a metal uneven structure 21 having a suitable shape on the electromagnetic wave enhancing element.
  • the dielectric concavo-convex structure 11 formed on the electromagnetic wave enhancing element may have a plurality of regions, and the dielectric concavo-convex structure 11 may be formed so as to have a different shape for each region.
  • the dielectric portion 1 shown in FIG. 4 has a dielectric concavo-convex structure 11 in a line-and-space shape, and the pitch of the convex portion of the dielectric concavo-convex structure 11 is increased toward the right region in the figure, and the lower region is formed. Regions 91 to 99 are formed in which the line width of the convex portion decreases as the distance increases.
  • the metal concavo-convex structure 21 is preferably bonded with a fixing substance 5 for fixing the substance as shown in FIG.
  • the metal part 2 may be surface-treated with a coupling agent or the like having a functional group capable of forming a chemical bond with the surface of the metal part 2 and capable of chemical bonding or chemical adsorption with a substance to be fixed.
  • the film thickness is preferably 5 nm or less, preferably 1 nm or less.
  • a silane having a functional group capable of binding or adsorbing to an amino group or a carboxyl group of an amino acid As a specific example, when producing the electromagnetic wave enhancing element of the present invention for amino acid analysis, as shown in FIG. 23, a silane having a functional group capable of binding or adsorbing to an amino group or a carboxyl group of an amino acid. What is necessary is just to perform the surface treatment of the metal part 2 with a coupling agent.
  • the water-soluble reagent 6 such as rhodamine may be repelled on the silver surface and may not reach a hot spot where Raman enhancement is strong.
  • a hydrophilic layer made of a material that is at least more hydrophilic than the metal part 2 may be formed on the surface of the metal part 2.
  • a hydrophilic layer 7 made of a hydrophilic dielectric material such as silicon oxide (SiO 2 )
  • SiO 2 silicon oxide
  • the hydrophilic layer 7 should be coated with a film thickness of 5 nm or less, preferably 1 nm or less.
  • the intermediate layer 3 may be any material that improves the adhesion between the dielectric portion 1 and the metal portion 2, and for example, platinum can be used.
  • the method for manufacturing an electromagnetic wave enhancing element according to the present invention includes a dielectric part forming step for forming a dielectric concavo-convex structure 11 on a dielectric part 1 and a metal part 2 constituting a metal concavo-convex structure 21 on the dielectric concavo-convex structure 11. And a metal part forming step to be formed.
  • the dielectric part forming step is for forming the dielectric concavo-convex structure 11 for controlling the shape and pitch of the metal concavo-convex structure 21.
  • the dielectric part forming step may be performed by any method as long as the predetermined dielectric uneven structure 11 can be formed. For example, a method of forming the dielectric concavo-convex structure 11 composed of a curved line and space in which the surface of the convex portion 14 bulges toward the concave portion will be described.
  • a base material 10 made of a dielectric is prepared.
  • a dielectric concavo-convex structure 15 is formed on the base material 10 by imprinting.
  • FIG. 6A a base material 10 made of a dielectric is prepared.
  • a dielectric concavo-convex structure 15 is formed on the base material 10 by imprinting.
  • the dielectric uneven structure 11 may be formed by performing UV ozone irradiation.
  • a resin is applied to the dielectric concavo-convex structure 15 to form a reverse arched mask 16 in the concave portion 22 of the dielectric concavo-convex structure 15.
  • the dielectric uneven structure 11 may be formed by performing UV ozone irradiation.
  • the concave portion 22 is formed such that the minimum width of the gap between adjacent convex portions forming the concave portion 22 is 25 nm or less, preferably 20 nm or less. Is good. As shown in FIG.
  • the recess 22 is preferably in contact with adjacent projections 24A and 24B forming the recess 22, but as long as the electric field can be sufficiently enhanced, FIG. As shown in (b), there may be a gap. Further, in FIG. 1A, there is no gap between the dielectric part 1 and the metal part 2, but if the electric field can be sufficiently enhanced, the dielectric part 1 and the metal part 2 can be seen as shown in FIG. There may be a gap 29 between them.
  • the metal part forming step may be any process as long as the metal part 2 constituting the metal uneven structure 21 can be formed. For example, a film forming technique such as sputtering, vapor deposition, or plating may be used.
  • the minimum value of ⁇ is 20 degrees or less, preferably 0 ° ⁇ . It is better to form the recess 22 so that ⁇ 15 °, more preferably 7 ° ⁇ ⁇ 13 °.
  • the minimum value of the angle ⁇ is 20 degrees or less, for example, as shown in FIG. 6E, a film forming technique that increases the vertical growth rate of the metal part 2, for example, What is necessary is just to form the metal part 2 using anisotropic sputtering. Moreover, as shown in FIG.6 (f), on the metal part 2 formed by sputtering, electroless plating may be given further and the shape of the metal part 2 may be adjusted. Thereby, the minimum value of the angle ⁇ can be adjusted.
  • the metal part forming step when the vertical thickness of the uppermost point 26 of the convex part 24 of the metal concavo-convex structure 21 is A and the vertical thickness of the lowermost point 23 is B, the metal is formed so that A> B.
  • Part 2 may be formed. More preferably, the metal part 2 is formed so that 0.9A> B, and more preferably, the metal part 2 is formed so that 0.6A> B.
  • the metal part forming step when the vertical thickness of the uppermost point 26 of the convex part 24 of the metal concave-convex structure 21 is A and half of the bottom width of the concave part 12 of the dielectric concave-convex structure C is A> C, You may form the metal part 2 so that it may become. More preferably, the metal part 2 is formed so that A> 2C, and more preferably, the metal part 2 is formed so that A> 3.5C. In this case, the size of C may be determined by adjusting the width of the concave portion of the dielectric concavo-convex structure 11 formed in the dielectric portion forming step.
  • the metal part forming step as shown in FIG. 22, first, a metal part main body for defining the metal uneven structure 21 is formed using a metal that can easily form the metal uneven structure 21. Next, a metal layer made of a metal different from the metal part body, for example, a metal having a higher electric field strength than the metal part body may be formed on the surface of the metal part body.
  • the metal layer is preferably 1 nm or more.
  • an electromagnetic wave enhancing element having a metal concavo-convex structure in which the convex portion and the concave portion are extended in one direction can be manufactured (see FIG. 7).
  • an intermediate layer for improving the adhesion between the dielectric part 1 and the metal part 2 on the dielectric part 1 between the dielectric part forming step and the metal part forming step. 3 may be included.
  • the intermediate layer 3 formation step may be performed by any method, and examples thereof include film formation techniques such as sputtering, vapor deposition, and plating.
  • the metal part 2 is utilized by using a film forming technique that increases the vertical growth rate of the metal part 2, for example, anisotropic sputtering. May be formed.
  • FIG.8 (c) you may adjust the shape of the metal part 2 by giving an electroless plating further on the metal part 2 formed by sputtering.
  • the metal part 2 may have a fixed substance binding step of binding a fixed substance capable of fixing a specific substance.
  • the metal part 2 may be surface-treated with a coupling agent or the like having a functional group capable of forming a chemical bond with the surface of the metal part 2 and capable of chemical bonding or chemical adsorption with a specific substance to be fixed.
  • the film thickness is preferably 5 nm or less, preferably 1 nm or less.
  • a silane having a functional group capable of binding or adsorbing to an amino group or a carboxyl group of an amino acid As a specific example, when producing the electromagnetic wave enhancing element of the present invention for amino acid analysis, as shown in FIG. 23, a silane having a functional group capable of binding or adsorbing to an amino group or a carboxyl group of an amino acid. What is necessary is just to perform the surface treatment of the metal part 2 with a coupling agent.
  • a water-soluble reagent such as rhodamine may be repelled on the silver surface and may not reach a hot spot where Raman enhancement is strong.
  • a hydrophilic layer 7 made of a material having a higher hydrophilic property than the metal part 2 such as silicon oxide (SiO 2 )
  • the reagent can reach the hot spot, and high sensitivity can be obtained.
  • the hydrophilic layer 7 should be coated with a film thickness of 5 nm or less, preferably 1 nm or less.
  • the electromagnetic wave enhancing element of the present invention will be described using simulation.
  • the software DiffractMOD manufactured by Synopsys, Inc. was used.
  • the electromagnetic wave enhancing element includes a dielectric part 1 in which a dielectric concavo-convex structure 11 in which the convex part 14 is a semicircle having a radius r, and a semicircle and a concentric circle that constitute the convex part 14.
  • a metal part 2 having a metal concavo-convex structure 21 composed of a convex part 24 which is a semicircle or a semi-ellipse obtained by enlarging it in the vertical direction was used.
  • the horizontal dimension from the center of the semicircle or semi-ellipse constituting the convex portion 24 is set to r + 10 nm, and the width d of the concave portion 12 of the dielectric concavo-convex structure 11 is changed to overlap the side portions of the adjacent convex portions 24.
  • the thickness B in the vertical direction of the lowest point 23 of the metal part 2 was adjusted.
  • pitches of the metal uneven structure 21 were simulated for every 10 nm from 100 nm to 200 nm.
  • PMMA is assumed as the material of the dielectric part 1
  • gold is assumed as the material of the metal part 2.
  • the maximum value Ex of the electric field enhancement exceeds 80 times when the angle ⁇ formed by the metal part 2 in the concave part 22 of the metal concave-convex structure 21 is 20 degrees or less.
  • the maximum value Ex of the electric field enhancement strength tends to be large when 0 ° ⁇ ⁇ 15 ° and further large when 7 ° ⁇ ⁇ 13 °.
  • the pitch P is at least 200 nm or less, there is a case where the maximum value Ex of the electric field enhancement exceeds 80 times, and the maximum value Ex of the electric field enhancement tends to increase as the pitch P decreases. .
  • the maximum value Ex of the electric field enhancement intensity was large when the pitch P was 100 ⁇ P ⁇ 120 nm.
  • FIG. 10 indicates that the maximum value Ex of the electric field enhancement strength tends to increase as the film thickness A is greater than the film thickness B.
  • the maximum value Ex of the electric field enhancement strength exceeds 80 times, and when the thickness ratio B / A is smaller than 0.6, It was found that the maximum value Ex of the electric field enhancement intensity exceeds 100 times.
  • the pitch P is at least 200 nm or less, there is a case where the maximum value Ex of the electric field enhancement exceeds 80 times, and the maximum value Ex of the electric field enhancement tends to increase as the pitch P decreases. .
  • the maximum value Ex of the electric field enhancement intensity was large when the pitch P was 100 ⁇ P ⁇ 120 nm.
  • FIG. 11 shows that the maximum value Ex of the electric field enhancement tends to increase as A is greater than C.
  • the ratio A / C of A and C is larger than 2
  • the maximum value Ex of the electric field strength exceeds 100 times
  • the ratio A / C is larger than 3.5
  • the maximum value Ex of the electric field strength increases. It turned out that there was a thing exceeding 120 times.
  • the pitch P is at least 200 nm or less
  • the maximum value Ex of the electric field enhancement exceeds 80 times, and the maximum value Ex of the electric field enhancement tends to increase as the pitch P decreases.
  • the maximum value Ex of the electric field enhancement intensity was large when the pitch P was 100 ⁇ P ⁇ 120 nm.
  • Example 1 Next, the electromagnetic wave enhancing element of the present invention was actually created, and the intensity of Raman scattered light (hereinafter referred to as Raman intensity) was confirmed. Samples 1 to 3 described later were used as the electromagnetic wave enhancing elements. Moreover, in order to compare with the electromagnetic wave enhancing element of the present invention, comparative samples 1 and 2 described later were used.
  • Example 1 Silver was formed at 200 W for 245 seconds by sputtering on a line-and-space dielectric concavo-convex structure having a height of 120 nm, a line width of 50 nm, and a pitch of 100 nm, and a concave portion with a smaller width was stretched in one direction toward the bottom side.
  • An electromagnetic wave enhancing element having a metal uneven structure was formed.
  • the thickness in the vertical direction of the uppermost point of the convex portion of the metal concavo-convex structure was 130 nm.
  • FIG. 12 shows a plan photograph and a sectional photograph of the electromagnetic wave enhancing element.
  • Example 2 Silver was formed at 200 W for 245 seconds by sputtering on a line-and-space dielectric concavo-convex structure having a height of 140 nm, a line width of 70 nm, and a pitch of 140 nm, and a concave portion with a smaller width was stretched in one direction toward the bottom side.
  • An electromagnetic wave enhancing element having a metal uneven structure was formed.
  • the thickness in the vertical direction of the uppermost point of the convex portion of the metal concavo-convex structure was 125 nm.
  • FIG. 13 shows a plan photograph and a sectional photograph of the electromagnetic wave enhancing element.
  • Example 3 Silver was formed at 200 W for 245 seconds by sputtering on a line-and-space dielectric concavo-convex structure having a height of 200 nm, a line width of 100 nm, and a pitch of 200 nm, and a concave portion with a smaller width was stretched in one direction toward the bottom side.
  • An electromagnetic wave enhancing element having a metal uneven structure was formed. The thickness in the vertical direction of the uppermost point of the convex portion of the metal concavo-convex structure was 127 nm.
  • a plane photograph and a cross-sectional photograph of the electromagnetic wave enhancing element are shown in FIG.
  • Electrode 1 Silver was formed at 200 W for 245 seconds by sputtering on a dielectric concavo-convex structure in which a cylinder having a height of 500 nm and a bottom diameter of 230 nm was arranged in a triangle with a pitch of 460 nm, thereby forming an electromagnetic wave enhancing element.
  • the thickness in the vertical direction of the uppermost point of the convex portion of the metal concavo-convex structure was 120 nm.
  • FIG. 15 shows a plan photograph and a sectional photograph of the electromagnetic wave enhancing element.
  • each electromagnetic wave enhancing element was irradiated with a laser having a wavelength of 532 nm perpendicularly at 3 mW for 1 second, and the Raman intensity at a Raman shift of 1430 cm ⁇ 1 was measured.
  • a laser was incident on the electromagnetic wave enhancing element (samples 1 to 3) of the present invention, and the azimuth angle and the Raman intensity at a Raman shift of 1430 cm ⁇ 1 were measured by rotating a vertical line at the incident point as a rotation axis. The result is shown in FIG.
  • the azimuth angle represents the Raman intensity every 22.5 degrees with the position indicating the maximum value of the Raman intensity of each electromagnetic wave enhancing element as 0 degree.
  • the Raman scattered light detection method using the electromagnetic wave enhancing element of the present invention includes an irradiation step of making light incident on the electromagnetic wave enhancing element, and an electromagnetic wave having a vertical line at the point where the light is incident on the electromagnetic wave enhancing element as a rotation axis. It is preferable to have a rotation step of rotating the enhancement element by at least 90 degrees and a measurement step of measuring the maximum intensity of the Raman scattered light of the electromagnetic wave enhancement element during the rotation step. Thereby, the electromagnetic wave enhancing element of the present invention can detect the characteristics of the subject at the position with the highest sensitivity.
  • the electromagnetic wave enhancing element of the present invention may have an alignment mark 4 indicating the extending direction of the concave portion of the metal part 2 as shown in FIG.
  • the alignment mark 4 may be formed anywhere in the electromagnetic wave enhancing element as long as the extending direction of the recess of the metal part 2 is known.
  • the alignment mark 4 can be formed on the dielectric part 1 or on the metal part 2.
  • FIG. 19 shows the maximum Raman intensity of each electromagnetic wave enhancing element (samples 1 to 3 and comparative samples 1 and 2).
  • the electromagnetic wave enhancing elements (samples 1 to 3) of the present invention having a metal concavo-convex structure having a shape in which a concave portion whose width decreases toward the bottom side is extended in one direction, the metal concavo-convex structure is cylindrical. It can be seen that the present invention shows a very large Raman intensity compared to the electromagnetic wave enhancing element (Comparative Sample 1) and the conical electromagnetic wave enhancing element (Comparative Sample 2).
  • Example 2 Next, the relationship between the minimum width of the gap between adjacent convex portions forming the concave portion of the metal concavo-convex structure and the Raman intensity was examined.
  • the electromagnetic wave enhancement element samples 4 to 6 described later were used.
  • Example 4 Silver was deposited at 200 W for 170 seconds by sputtering on a line-and-space dielectric concavo-convex structure having a height of 140 nm, a line width of 70 nm, and a pitch of 140 nm, and a concave portion with a smaller width was stretched in one direction toward the bottom side. An electromagnetic wave enhancing element having a metal uneven structure was formed. At this time, the minimum width of the gap between adjacent convex portions forming the concave portion of the metal concave-convex structure was 25.9 nm.
  • Example 5 Silver was deposited at 200 W for 195 seconds by sputtering on a line-and-space dielectric concavo-convex structure having a height of 140 nm, a line width of 70 nm, and a pitch of 140 nm, and a concave portion with a smaller width was stretched in one direction toward the bottom side. An electromagnetic wave enhancing element having a metal uneven structure was formed. At this time, the minimum width of the gap between adjacent convex portions forming the concave portion of the metal concavo-convex structure was 23.7 nm.
  • Example 6 Silver was formed at 200 W for 245 seconds by sputtering on a line-and-space dielectric concavo-convex structure having a height of 140 nm, a line width of 70 nm, and a pitch of 140 nm, and a concave portion with a smaller width was stretched in one direction toward the bottom side.
  • An electromagnetic wave enhancing element having a metal uneven structure was formed.
  • the minimum width of the gap between adjacent convex portions forming the concave portion of the metal concavo-convex structure was 20 nm.
  • Each electromagnetic wave enhancing element was irradiated with a laser having a wavelength of 532 nm perpendicularly at 3 mW for 1 second, and the maximum value of the Raman intensity at a Raman shift of 1430 cm ⁇ 1 was measured. The result is shown in FIG.
  • the electromagnetic wave enhancing element of the present invention increases the Raman intensity when the minimum width of the gap between adjacent convex portions forming the concave portion of the metal concave-convex structure is reduced. Specifically, it can be seen that the Raman intensity exceeds 10,000 when the minimum width of the gap is 25 nm or less, and exceeds 35000 when it is 20 nm or less.
  • the amino acid sequence determination method of the present invention includes a sequential decomposition step for sequentially decomposing amino acids from the N-terminus or C-terminus of a peptide or protein, a fractionation step for fractionating amino acids released by the sequential degradation step, and a fractionation step And an analysis step of analyzing the amino acid obtained by the method using the electromagnetic wave enhancing element of the present invention.
  • the sequential decomposition step may be any method as long as it is a method for releasing an amino acid located at the N-terminus or C-terminus of a peptide or protein.
  • a method using a protease a phenylisothiocyanate (PITC) is reacted with a free amino group at the N-terminal part of a peptide or protein to form a phenylthiocarbamyl derivative (PTC amino acid), and then anilinothiazolinone with trifluoroacetic acid
  • a method of releasing as (ATZ) -amino acid (Edman degradation) can be used.
  • an exopeptidase that hydrolyzes a peptide bond at the N-terminal or C-terminal of a peptide or protein and sequentially releases amino acids from the terminal can be used.
  • the exopeptidase may be used by mixing two or more types having different characteristics.
  • carboxypeptidase Y that exhibits high catalytic activity when the second or terminal residue from the terminal is an aromatic or aliphatic amino acid, and a terminal residue that is a basic amino acid
  • carboxypeptidase B showing high catalytic action can be used in combination.
  • the fragment after performing an endopeptidase treatment first to obtain a fragment having an appropriate length, the fragment may be subjected to degradation by exopeptidase.
  • the sequential decomposition step of the present invention it is preferable that only one residue is released from the end of the peptide or protein.
  • the Raman scattering signal is analyzed even when two or more amino acids are bound, and the type and order of the amino acids are determined. Can be identified. Therefore, in this case, it is possible to identify an amino acid even in the case where 2 or more residues are released in a bound state from the end of the peptide or protein in the sequential decomposition step of the present invention. In some cases.
  • the sequential decomposition may be performed using a column on which a protease, peptide, or protein is immobilized. Immobilization of protease, peptide or protein to the carrier packed in the column may be performed by any method. By using the immobilized column, a sample that has been subjected to the sequential decomposition step, that is, a sample containing free amino acids can be easily obtained at a desired timing.
  • the fractionation step in the amino acid sequence determination method of the present invention may be any method as long as it is a method for fractionating amino acids released by the sequential decomposition step.
  • the eluate from the column may be collected over time using a fraction collector or the like.
  • the activity of the protease when controlling the activity of the protease, only the eluate that has undergone the degradation reaction by the protease may be collected in correspondence with the control. Thereby, a desired amino acid can be efficiently recovered.
  • the analysis step in the amino acid sequence determination method of the present invention is a step of analyzing the amino acid obtained by the fractionation step using the electromagnetic wave enhancing element of the present invention, and is performed, for example, as follows.
  • the free amino acid obtained by the sorting step is fixed to the end of the dielectric layer of the electromagnetic wave enhancing element of the present invention.
  • the amino acid can be fixed to the end portion of the dielectric layer by physical adsorption to the surface of the end portion or by bonding via polybrene or a silane coupling agent.
  • a solution containing the amino acid obtained by the fractionation step onto the electromagnetic wave enhancing element of the present invention in which a silane coupling agent is bonded to the end of the dielectric layer in advance this is appropriately washed, It can be dried.
  • the electromagnetic wave enhancing element to which the amino acid is fixed is irradiated with an electromagnetic wave having a predetermined wavelength, and the obtained Raman scattering signal is analyzed to identify the fixed amino acid.
  • various analyzes are performed as necessary to determine the amino acid sequence.
  • Example 3 Next, a method for producing the electromagnetic wave enhancing element of the present invention will be described.
  • FIG. 21 when an inexpensive sputter film formation is performed on a line-and-space dielectric concavo-convex structure, since the migration of atoms is small in gold, the film formation shape must correspond to the arrival amount of atoms on a one-to-one basis. become. Then, when adjacent gold contacts, a gentle contact angle is taken, and the signal cannot be amplified by the metal part 2. On the other hand, as shown in FIG.
  • Example 4 Next, another method for producing an electromagnetic wave enhancing element will be described.
  • the water-soluble reagent 6 such as rhodamine is sometimes repelled on the Ag surface and does not reach the hot spot. In this case, no Raman enhancement occurs. Therefore, as shown in FIG. 25, a hydrophilic layer 7 having a film thickness of 5 nm or less, preferably 1 nm or less, is formed of a hydrophilic dielectric material such as SiO 2 on the surface of Ag (metal part 2). As a result, the reagent reaches the hot spot and high sensitivity is obtained.

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Abstract

Le but de la présente invention est de proposer un élément d'amplification des ondes électromagnétiques qui conduit à une amplification d'un champ électrique par rapport à des éléments classiques et qui rend possible une évaluation quantitative. L'élément d'amplification des ondes électromagnétiques comprend principalement : une partie diélectrique (1) dans laquelle est formée une structure diélectrique à évidements et saillies (11) ; et une partie métallique (2) possédant, sur la structure diélectrique à évidements et saillies (11), une structure métallique à évidements et saillies (21), dans laquelle une partie d'évidement (22) dont la largeur va en diminuant en allant vers sa face inférieure est allongée dans une direction. En outre, lorsqu'un angle entre des tangentes (25A, 25B) au niveau d'un point (23) présentant la même hauteur avec des parties en saillie voisines (24A, 24B) qui forment la partie d'évidement (22) est θ, la partie d'évidement (22) présente, de préférence, un θ d'une valeur minimale égale ou inférieure à 20 degrés.
PCT/JP2019/019810 2018-05-18 2019-05-17 Élément d'amplification des ondes électromagnétiques, son procédé de production, procédé de détection utilisant l'élément d'amplification des ondes électromagnétiques et procédé de détermination d'une séquence d'acides aminés WO2019221298A1 (fr)

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Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004097384A1 (fr) * 2003-04-25 2004-11-11 E2V Technologies (Uk) Limited Ensemble detecteur moleculaire
JP2008501982A (ja) * 2004-06-07 2008-01-24 ウェイン エー ワイマー 表面増強ラマン分光（ｓｅｒｓ）用基板表面の製造方法及びシステム、及びそれを用いた装置
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
JP2012063156A (ja) * 2010-09-14 2012-03-29 Seiko Epson Corp 光デバイスユニット及び検出装置
JP2013522639A (ja) * 2010-03-22 2013-06-13 アイメック 表面増強光検出のための方法およびシステム
JP2013195204A (ja) * 2012-03-19 2013-09-30 Seiko Epson Corp 試料分析基板および検出装置
JP2014037971A (ja) * 2012-08-10 2014-02-27 Hamamatsu Photonics Kk 表面増強ラマン散乱ユニット
WO2015136695A1 (fr) * 2014-03-14 2015-09-17 株式会社 東芝 Dispositif et procédé de détection modulaire

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004097384A1 (fr) * 2003-04-25 2004-11-11 E2V Technologies (Uk) Limited Ensemble detecteur moleculaire
JP2008501982A (ja) * 2004-06-07 2008-01-24 ウェイン エー ワイマー 表面増強ラマン分光（ｓｅｒｓ）用基板表面の製造方法及びシステム、及びそれを用いた装置
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
JP2013522639A (ja) * 2010-03-22 2013-06-13 アイメック 表面増強光検出のための方法およびシステム
JP2012063156A (ja) * 2010-09-14 2012-03-29 Seiko Epson Corp 光デバイスユニット及び検出装置
JP2013195204A (ja) * 2012-03-19 2013-09-30 Seiko Epson Corp 試料分析基板および検出装置
JP2014037971A (ja) * 2012-08-10 2014-02-27 Hamamatsu Photonics Kk 表面増強ラマン散乱ユニット
WO2015136695A1 (fr) * 2014-03-14 2015-09-17 株式会社 東芝 Dispositif et procédé de détection modulaire

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
CIALLA, DANA ET AL: "SERS-based detection of biomolecules", NANOPHOTONICS, vol. 3, no. 6, 1 January 2014 (2014-01-01), pages 383 - 411, XP055653477, ISSN: 2192-8606, DOI: 10.1515/nanoph-2013-0024 *

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