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WO2023281674A1 - Capteur de tension de surface de type film et procédé de fabrication de capteur de tension de surface de type film - Google Patents

Capteur de tension de surface de type film et procédé de fabrication de capteur de tension de surface de type film Download PDF

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Publication number
WO2023281674A1
WO2023281674A1 PCT/JP2021/025694 JP2021025694W WO2023281674A1 WO 2023281674 A1 WO2023281674 A1 WO 2023281674A1 JP 2021025694 W JP2021025694 W JP 2021025694W WO 2023281674 A1 WO2023281674 A1 WO 2023281674A1
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WIPO (PCT)
Prior art keywords
film
sensor
layer
membrane
silicon
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PCT/JP2021/025694
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English (en)
Japanese (ja)
Inventor
賢司 宮崎
久 萩原
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日本電気株式会社
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Priority to PCT/JP2021/025694 priority Critical patent/WO2023281674A1/fr
Priority to JP2023532962A priority patent/JPWO2023281674A1/ja
Publication of WO2023281674A1 publication Critical patent/WO2023281674A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N19/00—Investigating materials by mechanical methods
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N5/00—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid
    • G01N5/02—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid by absorbing or adsorbing components of a material and determining change of weight of the adsorbent, e.g. determining moisture content

Definitions

  • the present invention relates to a membrane-type surface stress sensor and a method for manufacturing the membrane-type surface stress sensor.
  • Target detection is important in a wide variety of fields such as food and medicine, and various methods have been proposed.
  • a membrane-type surface stress sensor has attracted attention (see Patent Document 1).
  • the film-type surface stress sensor can analyze the presence and amount of the target by, for example, binding a target to a film such as a silicon film, deforming the film, and measuring variations in electrical resistance caused by the deformation. .
  • the present inventors invented MSS in which a binding substance such as an aptamer capable of binding to a target is immobilized on the surface of the MSS membrane.
  • the inventors have found a problem that the immobilization efficiency of the binding substance on the membrane of MSS is low.
  • the second object of the present invention is to provide an MSS capable of improving the binding substance immobilization efficiency.
  • the membrane is a membrane that deforms in response to surface stress
  • the sensor substrate comprises a support area and circuitry; the support region supports the membrane and comprises a piezoresistive element;
  • the piezoresistive element is an element that detects deformation of the film,
  • the circuit is connected to the piezoresistive element,
  • the circuit includes a metal capable of forming an oxide film by oxidation treatment,
  • the circuit has an oxide film layer on its surface.
  • a method for manufacturing a film-type surface stress sensor according to the present invention is a film-type stress sensor comprising a film and a circuit, wherein the circuit includes a metal capable of forming an oxide film by oxidation treatment. and forming an oxide film layer on the surface of the circuit by oxidizing the circuit.
  • the membrane is a membrane that deforms in response to surface stress
  • the sensor substrate comprises a support area; the support region supports the membrane and comprises a piezoresistive element;
  • the piezoresistive element is an element that detects deformation of the film,
  • the film comprises an amorphous layer.
  • a method for manufacturing a film-type surface stress sensor of the present invention is a film-type stress sensor comprising a film and a circuit, wherein the film is subjected to an amorphization treatment (amorphization process). quenching treatment) to form an amorphous layer on the surface of the film.
  • a circuit containing a metal capable of forming an oxide film by oxidation treatment is subjected to oxidation treatment to form an oxide film layer on the surface of the circuit. and and forming an amorphous layer on the surface of the film by subjecting the film to amorphization treatment.
  • the target analysis method of the present invention includes an applying step of applying a voltage to a membrane surface stress sensor in a sample liquid; an analysis step of analyzing the target in the sample liquid by measuring the stress change of the piezoresistive element in the film-type surface stress sensor;
  • the film-type surface stress sensor is the first film-type surface stress sensor of the present invention or the second film-type surface stress sensor of the present invention.
  • the first MSS of the present invention it is possible to insulate between circuits.
  • the second MSS of the present invention it is possible to improve the immobilization efficiency of the binding substance on the MSS membrane.
  • FIG. 5 is a schematic diagram showing an example of a method for producing MSS according to Embodiment 2.
  • FIG. 6 is a schematic cross-sectional view showing another example of the silicon film of the MSS in Embodiment 2.
  • FIG. 7 is a schematic diagram showing another example of the method for producing MSS according to Embodiment 2.
  • FIG. 8 is a schematic diagram showing an example of the configuration of the MSS according to the third embodiment.
  • 9 is a photograph showing a transmission electron microscope image of the silicon film of MSS of Embodiment 3 in Example 1.
  • the term "membrane-type surface-stress sensor (MSS)” refers to a sensor in which a film that deforms in response to surface stress is supported by a support having a piezoresistive element. do.
  • MSS membrane-type surface-stress sensor
  • the film when the film receives stress, the film deforms (generates strain) due to the generation of strain or the like. Then, according to the amount of deformation of the film, stress is generated in the piezoresistive element of the support supporting the film, and the resistance value of the piezoresistive element changes in proportion to the stress.
  • the presence or absence of the target bound to the membrane can be qualitatively analyzed indirectly by applying a voltage to the MSS and measuring an electrical signal accompanying a change in resistance value. can.
  • the MSS sensor by applying a voltage to the MSS and measuring an electrical signal accompanying a change in resistance value, it is possible to quantitatively analyze the amount of the target that has contributed to the generation of stress in the film. .
  • the "target” is not particularly limited and can be set arbitrarily.
  • Said target may be, for example, a substance that can come into contact with said membrane or said binding substance in a liquid, ie in liquid phase.
  • the target has, for example, one or more regions to which the binding substance binds, ie epitopes of the binding substance.
  • the targets include, for example, microorganisms including bacteria such as anthrax, Escherichia coli, salmonella, and tubercle bacillus; viruses such as SARS-CoV-2 and influenza virus; allergens; Examples of the allergen include grains such as wheat; eggs; meat; fish; shellfish; vegetables;
  • the type of the target is not particularly limited, and examples thereof include high-molecular compounds such as proteins, sugar chains, nucleic acids, and polymers; low-molecular-weight compounds; and the like.
  • antibody can also be referred to as a soluble immunoglobulin that has binding properties to a target.
  • Types of the antibody include, for example, IgA, IgD, IgE, IgG, or IgM.
  • IgA includes, for example, IgA1 or IgA2.
  • IgG includes, for example, IgG1, IgG2, IgG3, or IgG4.
  • the antibody may be an antigen-binding fragment thereof, that is, a partial peptide of the antibody that has binding properties to the target. Said antigen-binding fragment is for example a polypeptide comprising a part of said antibody, more particularly the binding or variable region of said antibody.
  • the antibody As the antibody, a known antibody or an antigen-binding fragment thereof having a binding property to a target may be used, or a new antibody or an antigen-binding fragment thereof obtained by immunizing an animal or the like with a target may be used. good.
  • the antibody may be a monoclonal antibody or a polyclonal antibody.
  • the antibody may be a blood-derived fraction such as serum or plasma that contains antibodies capable of binding to the target.
  • an "aptamer” is a nucleic acid molecule that has binding properties to a target.
  • the aptamer can also be referred to as, for example, a nucleic acid molecule that specifically binds to a target.
  • the constituent units of the aptamer are, for example, nucleotide residues and non-nucleotide residues.
  • Said nucleotide residues include, for example, deoxyribonucleotide residues and ribonucleotide residues, and said nucleotide residues may, for example, be modified or unmodified.
  • the aptamer examples include DNA aptamers composed of deoxyribonucleotide residues, RNA aptamers composed of ribonucleotide residues, aptamers containing both, aptamers containing modified nucleotide residues, and the like.
  • the length of the aptamer is not particularly limited, and is, for example, 10-200 bases.
  • an existing aptamer may be used, or an aptamer newly obtained using, for example, the SELEX method or the like may be used depending on the target.
  • the binding substance is preferably an aptamer or an antibody.
  • the binding substance may be labeled.
  • the label is not particularly limited, and examples thereof include fluorescent substances, dyes, isotopes, enzymes, and the like.
  • the fluorescent substance include pyrene, TAMRA, fluorescein, Cy3 dye, Cy5 dye, FAM dye, rhodamine dye, Texas Red dye, and fluorophores such as JOE, MAX, HEX, and TYE.
  • Alexa dyes such as Alexa488 and Alexa647.
  • the enzyme include luciferase, alkaline phosphatase, peroxidase, ⁇ -galactosidase, glucuronidase and the like.
  • the binding substance is a nucleic acid
  • the label is bound, for example, to at least one of the 5' end and the 3' end of the nucleic acid.
  • the label and carrier are, for example, attached to the N-terminus, C-terminus or side chain of the protein.
  • the label is, for example, directly or indirectly bound to the binding substance.
  • said label is attached via a linker.
  • the "sample liquid” may be any liquid.
  • the specimen to be collected is liquid, it may be used as a liquid sample as it is, or may be a liquid sample prepared by diluting, suspending, dispersing, or the like with a liquid solvent.
  • the specimen to be collected is solid, it may be a liquid sample prepared by dissolving, suspending, dispersing, or the like in a liquid solvent.
  • the specimen to be collected is a gas, for example, it may be a liquid sample obtained by concentrating an aerosol in the gas, or a liquid sample prepared by dissolving, suspending, or dispersing in a liquid solvent.
  • the type of the liquid solvent is not particularly limited.
  • the sample liquid may be, for example, a liquid containing targets, a liquid containing no targets, or a liquid whose presence or absence of targets is unknown.
  • FIG. 1 is a schematic diagram showing an MSS 1 of Embodiment 1.
  • FIG. 1 (A) is a schematic plan view of the MSS 1 and a schematic enlarged view of the periphery of the circuit 12A, and (B) is a schematic cross-sectional view seen from the II direction in (A).
  • the MSS 1 of Embodiment 1 comprises a sensor substrate 10, electrodes 11, aluminum wires 12A (circuits), MSS films 13 (films), piezoresistive elements 14, and support regions 15.
  • FIG. 1 is a schematic diagram showing an MSS 1 of Embodiment 1.
  • FIG. 1 (A) is a schematic plan view of the MSS 1 and a schematic enlarged view of the periphery of the circuit 12A
  • (B) is a schematic cross-sectional view seen from the II direction in (A).
  • the MSS 1 of Embodiment 1 comprises a sensor substrate 10, electrodes 11, aluminum wires 12A (circuits), MSS films 13 (films), piezoresistive
  • the aluminum wire 12A is composed of a conductive layer 121 and an insulating layer 122 (oxide layer).
  • the MSS film 13 is supported by the sensor substrate via a support region 15 having a piezoresistive element 14 formed thereon. Also, the MSS film 13 is connected to the aluminum wire 12A through the piezoresistive element 14, and the aluminum wire 12A is connected to the electrode 11 respectively.
  • the aluminum wire 12A constitutes a Wheatstone bridge circuit including four piezoresistive elements 14 .
  • the MSS 1 of Embodiment 1 includes the electrode 11, the electrode 11 has an arbitrary configuration and may or may not be present. If the MSS 1 does not have the electrode 11, the aluminum wire 12A is connected to the voltage applying device.
  • the sensor substrate 10 is a substrate on which various configurations of the MSS 1 such as the electrodes 11, aluminum wires 12A, and the MSS film 13 can be arranged.
  • a semiconductor substrate can be used, and as a specific example, a wafer made of silicon can be used.
  • the sensor substrate 10 supports the MSS film 13 by means of support regions 15 .
  • the sensor substrate 10 preferably partially supports the MSS film 13 , and specifically preferably partially supports the side surfaces of the MSS film 13 .
  • the sensor substrate 10 supports the MSS film 13 via four supporting regions 15 (supporting portions), but the present invention is not limited to this, and the sensor substrate 10 may be any A number of support regions 15 may support the MSS membrane 13 .
  • the sensor substrate 10 supports one MSS film with four support regions 15, but the present invention is not limited to this. Multiple support regions 15 may each support an MSS membrane 13 . In this case, the number of support regions 15 and the number of supported MSS films 13 on one sensor substrate 10 are not particularly limited, and each may be one or two or more.
  • the electrode 11 can be configured to apply a voltage to the MSS1 by being connected to a voltage applying device outside the MSS1.
  • the electrode 11 thereby makes it possible to measure the change in stress on the piezoresistive element 14 as a voltage value.
  • the material of the electrode 11 may be any conductive metal, and the description of the material of the circuit in the description of the aluminum wire 12A, which will be described later, can be used.
  • the material of the electrodes 11 may be the same as or different from that of the circuit.
  • the electrode 11 is composed of a conductive layer made of a conductive metal, but the present invention is not limited to this. may be formed.
  • the aluminum wire 12A electrically connects the electrode 11 and the piezoresistive element 14, and makes it possible to measure the change in stress on the piezoresistive element 14 as a voltage value when voltage is applied by the voltage application device outside the MSS1.
  • the aluminum wire 12A has a conductive layer 121 laminated on the substrate 10 on the center side thereof, and the conductive layer 121 is covered with an insulating layer 122. As shown in FIG. That is, the conductive layer 121 is sealed between the substrate 10 and the insulating layer 122 . Therefore, the conductive layer 121 is insulated from the outside of the aluminum wire 12A.
  • the insulation between the aluminum wires 12A is ensured in the MSS1, short-circuiting between the aluminum wires 12A can be suppressed even if the MSS1 is immersed in the sample liquid and a voltage is applied to the MSS1.
  • the conductive layer 121 is made of aluminum that exhibits conductivity, and more specifically, is made of non-oxidized aluminum.
  • the insulating layer 122 is made of aluminum exhibiting insulating properties, specifically, made of aluminum oxide.
  • the insulation means insulation to the extent that the short circuit between the circuits is significantly suppressed when the MSS 1 is used under normal conditions, as compared with the MSS without the insulation layer 122 .
  • the material of the circuit is aluminum, but in the present invention, the material of the circuit is not limited to this, and may be any metal capable of forming an oxide film layer by oxidation treatment.
  • metals capable of forming the oxide film layer include valve metals, and specific examples include aluminum, tantalum, niobium, titanium, hafnium, zirconium, zinc, tungsten, bismuth, antimony, molybdenum, chromium, iron, or alloys thereof.
  • the MSS 1 can ensure insulation between circuits without forming an insulating layer using resin.
  • the width and height of the conductive layer 121 are sufficient as long as they are thick enough to electrically connect the electrodes 11 and the piezoresistive elements 14 .
  • the width of the conductive layer 121 is, for example, 1-100 ⁇ m, 10-20 ⁇ m.
  • the height of the conductive layer 121 is, for example, 0.1 to 100 ⁇ m, 1 to 20 ⁇ m.
  • the thickness of the insulating layer 122 is such that when the MSS 1 is used under normal conditions, compared to the MSS without the insulating layer 122, the insulating property is such that the short circuit between the aluminum wires 12A is significantly suppressed. Any thickness can be used.
  • the thickness of the insulating layer 122 is, for example, 0.5 to 50 nm, 1 to 10 nm.
  • the aluminum wire 12A forms a Wheatstone bridge circuit with the piezoresistive element 14, but the circuit structure of the aluminum wire 12A is not limited to this, and the change in stress on the piezoresistive element 14 can be electrically measured. circuit structure.
  • the MSS film 13 deforms according to the surface stress, and the deformation applies stress to the piezoresistive element 14 .
  • the material of the MSS film 13 is not particularly limited as long as it deforms in response to surface stress and applies stress to the piezoresistive element 14 due to the deformation.
  • the MSS film 13 is, for example, a thin film, and its thickness and surface area are not particularly limited, and are similar to MSS films used in commercially available MSS, for example.
  • the planar shape of the MSS film 13 is circular, the present invention is not limited to this, and may be any shape.
  • the planar shape of the MSS film 13 is preferably a perfect circle because it can increase distortion due to surface stress.
  • the material of the MSS film 13 is not particularly limited, and is silicon, for example.
  • a silicon film can be used as the MSS film 13, and as a specific example, any silicon film can be used regardless of p-type or n-type polarity.
  • Si(100) can be used.
  • the piezoresistive element 14 is an element that detects deformation of the MSS film 13 .
  • the stress change is concentrated in support region 15 . Therefore, in the MSS 1 , the piezoresistive element 14 is formed in the supporting region 15 at a location supporting the MSS film 13 .
  • the piezoresistive element 14 changes its resistance value according to the stress change. Therefore, in the MSS1, by applying a voltage through the electrode 11 and the aluminum wire 12A, an output voltage corresponding to the stress can be obtained due to the change in the resistance value. This allows the MSS 1 to analyze the presence or amount of the target in the sample liquid.
  • the piezoresistive element 14 can be manufactured, for example, by doping an arbitrary region of the support region 15 made of a silicon film to make it p-type by doping an impurity. Therefore, the piezoresistive element 14 preferably uses p-type Si formed on a silicon film. Note that the piezoresistive element 14 is formed at the location where the support region 15 supports the MSS film 13, but may be formed in the vicinity of that location.
  • FIG. 2 is a schematic cross-sectional view showing an example of a method for manufacturing the MSS1.
  • MSS is prepared.
  • MSS commercially available MSS may be used, or self-prepared.
  • the MSS 1 forms the piezoresistive element 14 by, for example, connecting the MSS film 13 to the supporting region 15 of the sensor substrate 10 and then doping the supporting region 15 .
  • the manufacturing method of the first embodiment can form the electrodes 11 and the aluminum wires 12A, which are circuits, by wiring aluminum by sputtering, vapor deposition, or the like.
  • the aluminum wire 12A formed on the MSS is made of conductive aluminum, that is, non-oxidized aluminum. Therefore, in the manufacturing method of the first embodiment, the insulating layer 122 is formed on the surface of the aluminum wire 12A. Specifically, in the manufacturing method of the first embodiment, the aluminum wire 12A is oxidized. In the manufacturing method of Embodiment 1, the oxidation treatment oxidizes the aluminum on the surface of the aluminum wire 12A, thereby forming an insulating oxide film as shown in FIG. is formed.
  • the oxidation treatment may be any treatment as long as it causes an oxidation reaction with a metal such as aluminum. be done.
  • the acid can be applied to the surface of the aluminum wire 12A, and the aluminum wire 12A and the acid can be reacted for a predetermined time.
  • the acid used for the acid treatment is an acid that can act as an oxidizing agent, and specific examples include nitric acid, sulfuric acid, phosphoric acid, oxalic acid, chromic acid, etc., preferably concentrated nitric acid, concentrated sulfuric acid, concentrated phosphoric acid, etc. acid, concentrated oxalic acid, concentrated chromic acid.
  • the reaction time with the acid in the acid treatment can be set according to the type of the acid and the metal forming the circuit, and is, for example, 15 to 180 minutes, 60 to 90 minutes.
  • the reaction temperature of the acid treatment is, for example, 0 to 50°C, preferably 30°C or less.
  • the plasma treatment can be performed by placing the MSS1 in a plasma generator and generating plasma while supplying oxygen to the plasma generator.
  • the oxygen concentration in the gas supplied to the plasma generator may be, for example, the default setting of each device, eg, 80-100%, 85-95%, about 90%.
  • the plasma treatment time can be set according to the type of metal forming the circuit and the thickness of the insulating layer 122 to be formed. One hour.
  • an insulating layer 122 containing aluminum oxide and aluminum hydroxide is formed on the surface of the aluminum wire 12A. Therefore, in the manufacturing method of Embodiment 1, after the oxygen plasma treatment, the formed aluminum hydroxide is preferably converted into aluminum oxide by a step including a heat treatment such as an annealing step to be described later. Thereby, the manufacturing method of Embodiment 1 can improve the insulating property of the formed insulating layer 122 .
  • the manufacturing method of Embodiment 1 can manufacture MSS1.
  • the sample liquid is brought into contact with the MSS1 (contact step).
  • the conditions for immersing MSS1 in the sample liquid are not particularly limited, and examples thereof include a temperature of 20 to 35° C. and a temperature of 0.1 to 120 minutes and a temperature of 50 to 60° C. and 0.1 to 120 minutes.
  • the MSS1 has a plurality of MSS membranes, for example, the plurality of MSS membranes in the MSS1 may be immersed in the same sample liquid at the same time.
  • a voltage is applied to the MSS1 in the sample liquid (liquid phase) (application step).
  • Conditions for applying the voltage are not particularly limited, and for example, conditions similar to those for commercially available MSS can be exemplified.
  • the liquid phase may be, for example, the sample liquid in the contacting step, or may be another solvent. In the latter case, the MSS1 after the contacting step is removed from the sample solution, immersed in a new solvent, and voltage is applied.
  • the solvent is not particularly limited, and examples thereof include PBS, buffer solutions such as Tris-HCl, water, and the like.
  • the timing of the application is the start of the contact process or after a predetermined time has elapsed since the start of the contact process.
  • the target in the sample liquid is analyzed by measuring the stress change of the piezoresistive element 14 in MSS1.
  • the stress change can be measured, for example, by measuring electrical signals, and a commercially available measurement module (eg, MSS-8RM, NANOSENSOR) can be used.
  • a reference measurement value may be obtained using a control sample solution that does not contain the target.
  • the target may be analyzed using a reference measurement value and a measurement value (signal) of the sample liquid.
  • an insulating layer 122 is formed on the surface of the aluminum wire 12A, which is the line. Therefore, in the MSS1, since the insulation between the aluminum wires 12A is ensured, it is possible to perform measurement in a liquid phase such as a sample liquid. Therefore, MSS1 can be suitably used for the analysis of liquid samples.
  • an oxide film layer can be formed by oxidation treatment, and the oxide film layer is used as the insulating layer 122 . be able to.
  • the manufacturing method according to the first embodiment does not require coating of the lines with resin, and damage due to adhesion of the resin to the piezoresistive element 14 can be avoided. Therefore, according to the manufacturing method of Embodiment 1, the breakage rate during manufacturing can be reduced, so the yield in manufacturing the MSS1 can be improved.
  • the manufacturing method of Embodiment 1 one type of oxidation treatment is performed, but the present invention is not limited to this, and multiple oxidation treatments may be performed. Moreover, the manufacturing method of Embodiment 1 may further include a step of annealing the formed oxide film layer. Another example of the manufacturing method of Embodiment 1 will be described with reference to FIG.
  • FIG. 3 is a schematic diagram showing another example of the method for manufacturing the MSS1 of Embodiment 1.
  • the conductive layer 121 of the aluminum wire 12A is subjected to two-stage oxidation treatment (FIGS. 3B and 3C), and then heat treatment to remove the oxide formed. Annealing the film layer forms an insulating layer 122 having a higher insulating property (FIG. 3A).
  • the MSS after the acid treatment is subjected to plasma treatment in an oxygen atmosphere.
  • the oxide film layer formed on the surface of the aluminum wire 12A is grown to increase the thickness of the insulating layer 122.
  • aluminum hydroxide is formed in the insulating layer 122 in addition to aluminum oxide.
  • the manufacturing method of another example of Embodiment 1 heat treatment is performed on the MSS after the oxygen plasma treatment.
  • the insulating layer 122 formed on the surface of the aluminum wire 12A is annealed, and the aluminum hydroxide in the insulating layer 122 is removed.
  • the lower limit of the temperature in the heat treatment is preferably 100° C. or higher, 200° C. or higher, or 300° C. or higher, for example.
  • the upper limit of the temperature in the heat treatment is, for example, 400° C. or less.
  • the temperature in the heat treatment is, for example, 100 to 400°C, 200 to 400°C, 300 to 400°C, and about 350°C.
  • the heat treatment time is, for example, 60 to 180 minutes.
  • the manufacturing method of another example of Embodiment 1 can manufacture MSS1.
  • the thickness of the insulating layer 122 in the MSS1 can be increased. Therefore, according to the manufacturing method of another example of the first embodiment, the MSS1 with higher insulation can be manufactured.
  • the binding substance is not immobilized on the MSS membrane 13 .
  • the present invention is not limited to this, and the binding substance may be immobilized on MSS1.
  • the binding substance may be immobilized on one surface of the MSS membrane 13, or may be immobilized on both surfaces.
  • the binding substance on one surface and the binding substance on the other surface are preferably the same binding substance that binds to the same target, for example.
  • the present invention is not limited to this, and a protein such as an antibody may be immobilized. good too.
  • the surface of the MSS film 13 may be, for example, one surface or both surfaces.
  • the method for immobilizing the aptamer on the MSS membrane 13 is not particularly limited, and the aptamer may be immobilized directly or indirectly on the MSS membrane 13.
  • the MSS membrane 13 and the aptamer can be immobilized by covalent bonding or the like by chemically treating the aptamer.
  • the direct fixing method includes, for example, a method using photolithography, and for specific examples, reference can be made to US Pat. No. 5,424,186.
  • Another direct immobilization method is, for example, a method of synthesizing the aptamer on the MSS membrane 13 . This method includes, for example, a so-called spot method, and for specific examples, reference can be made to US Pat. No. 5,807,522.
  • the aptamer can be immobilized on the MSS membrane 13 via a linker.
  • the type of linker is not limited at all, and examples thereof include a combination of biotin or a biotin derivative (hereinafter referred to as biotins) and avidin or an avidin derivative (hereinafter referred to as avidins).
  • biotins biotin derivative
  • avidins avidin derivative
  • examples of the biotin derivative include biocytin
  • examples of the avidin derivative include streptavidin.
  • the length of the linker is, for example, the length of the shortest molecular chain (main chain long).
  • the main chain length of the linker is 1 to 20, and is preferably 1 to 15, 1 to 13, 3 to 13, 5 to 13, 1 to 11, 3 to 11 because it can improve the sensitivity of the MSS membrane 13. , 1-10, 3-10, 1-8, 3-8, 1-5, 1-3, 1 or 2. Examples of immobilization methods are shown below, but the present invention is not limited to these.
  • the biotins are bound to one of the MSS membrane 13 and the aptamer, and the avidins are bound to the other. Then, the aptamer can be indirectly immobilized on the MSS membrane 13 by binding the biotins and the avidins.
  • the aptamer was indirectly immobilized on the MSS membrane by utilizing the specific binding between avidins and biotins, that is, the affinity of biotins to avidins.
  • affinity tags other than avidins-biotins may be used.
  • the affinity tag include His tag (His ⁇ 6 tag)-nickel ion, glutathione-S-transferase-glutathione, maltose binding protein-maltose, epitope tag (myc tag, FLAG tag, HA (hemagglutinin) tag)- Antibodies or antigen-binding fragments can be used.
  • His tag His ⁇ 6 tag
  • maltose binding protein-maltose epitope tag
  • epitope tag myc tag, FLAG tag, HA (hemagglutinin) tag
  • Antibodies or antigen-binding fragments can be used.
  • the point that other affinity tags may be used is
  • the aptamer may be immobilized on the MSS membrane 13 via an intervening membrane, for example.
  • the intervening film is, for example, a film of metal such as gold, and can be formed by vapor-depositing the metal on the MSS film 13 .
  • the thickness of the intervening film is not particularly limited, and is, for example, 10 to 100 nm.
  • the intervening film may be, for example, one layer or two or more layers.
  • the intervening film is, for example, two-layered, and a metal film (adhesive film) for adhesion to the MSS film 13 is provided in order to improve the adhesiveness of the gold film. It is preferable to form the gold film through the metal.
  • the metal of the adhesive film include titanium and chromium.
  • the thickness of the adhesive film is, for example, 0.1 to 10 nm, and the thickness of the gold film is, for example, 0.1 to 100 nm.
  • biotins When the biotins are bound to the intervening membrane, for example, self-assembled monolayers (SAMs) of thiolalkanes are formed on the surface of the intervening membrane using thiolalkanes to which the biotins are bound.
  • SAMs self-assembled monolayers
  • the aptamer formed and bound with the biotins may be brought into contact, and the aptamer may be immobilized by binding the biotins and the avidins.
  • the MSS film 13 is reacted with a silane coupling agent having an amino group to bond the amino group onto the MSS film 13 .
  • the reaction can be carried out, for example, by coating the MSS film 13 with a solution containing a silane coupling agent having an amino group.
  • the conditions for the cross-linking reaction can be appropriately determined according to, for example, the type of cross-linking agent.
  • the avidins are bound to the other end of the cross-linking agent such as glutaraldehyde.
  • the surface of the MSS membrane is washed, and a solution containing avidins is applied to bind the other end of the cross-linking agent to the main chain or side chain of amino acids of avidins.
  • the biotin-bound aptamer is brought into contact with the MSS membrane 13 treated in this way, and the aptamer can be immobilized by binding the biotins and the avidins.
  • Silane coupling agents having an amino group include, for example, N-(2-aminoethyl)-3-aminopropylmethyldimethoxysilane (eg, KBM-602 (manufactured by Shin-Etsu Silicone Co., Ltd.)), N-(2-aminoethyl )-3-aminopropyltrimethoxysilane (e.g., KBM-603 (manufactured by Shin-Etsu Silicone Co., Ltd.)), 3-aminopropyltrimethoxysilane (e.g., KBM-903 (manufactured by Shin-Etsu Silicone Co., Ltd.)), 3-aminopropyltriethoxy Silane (e.g., KBE-903 (manufactured by Shin-Etsu Silicone Co., Ltd.)), 3-(2-aminoethylamino)propyltrimethoxysilane (e.g., GEN
  • the cross-linking agent can be appropriately determined according to the functional group of the main chain or side chain of the amino acid that binds to the linker.
  • the functional group include an amino group (--NH 2 ), a thiol group (--SH), a carboxyl group (--COOH) and the like.
  • Said amino groups are present, for example, at the N-terminus of proteins or peptides or at the side chains of lysines.
  • the thiol group is, for example, a side chain of cysteine.
  • Said carboxyl groups are present, for example, at the C-terminus of proteins or peptides or at the side chains of aspartic acid or glutamic acid.
  • examples of the cross-linking agent include a cross-linking agent having aldehyde groups at both ends such as glutaraldehyde; bis(sulfosuccinimidyl) suberate (BS3); Disuccinimidyl glutarate (DSG), Disuccinimidyl suberate (DSS), Dithiobis(succinimidylpropionate), Dithiobis(sulfosuccinimidylpropionate) (DSP), Dithiobis(succinimidylpropionate) (DTSP) , dithiobis(sulfosuccinimidyl propionate) (DTSSP), disuccinimidyl tartrate (DST), ethylene glycol bis(succinimidyl succinate) (ESG), ethylene glycol bis(sulfosuccinimidyl succinate) ) (Sulfo-ESG), N
  • examples of the cross-linking agent include N-(6-maleimidocaproyloxy) succinimide (EMCS), N-(6-maleimidocaproyloxy) sulfosuccinimide (Sulfo -EMCS), N-(8-maleimidocaryloxy) succinimide (HMCS), N-(8-maleimidocaryloxy) sulfosuccinimide (Suflo-HMCS), N- ⁇ -maleimidoacet-oxysuccinimide ester (AMAS), N- ⁇ -maleimidopropyl-oxysuccinimide ester (BMPS), N- ⁇ -maleimidobutyryl-oxysuccinimide ester (GMBS), N- ⁇ -maleimidobutyryl-oxysulfosuccinimide ester (Sulfo-GMBS), m-maleimidobenzoyl-N-hydroxy
  • examples of the cross-linking agent include dicyclohexylcarbodiimide (DCC), 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), N-hydroxysuccinimide (NHS ), N-hydroxysulfosuccinimide (Sulfo-NHS), acetic anhydride, and the like.
  • DCC dicyclohexylcarbodiimide
  • EDC 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide
  • NHS N-hydroxysuccinimide
  • Sulfo-NHS N-hydroxysulfosuccinimide
  • acetic anhydride and the like.
  • the cross-linking agent is preferably a cross-linking agent that does not substantially cause self-condensation because the length of the linker can be made substantially constant or constant.
  • the above-mentioned "constant linker length" means, for example, that in the linkers of a plurality of aptamers, the linkers of each aptamer have substantially the same or the same length.
  • the length of the linker can be achieved, for example, by making the structures of the linkers the same.
  • the sensitivity of MSS can be improved by using such a cross-linking agent. The improvement in sensitivity is presumed to be due to the following reasons.
  • the present invention is not limited to the following estimation.
  • a target When a target binds to an aptamer, steric hindrance caused by the target occurs around the aptamer bound to the target. If the aptamers are immobilized at different distances from the MSS membrane 13, the targets are more likely to come into contact with aptamers that are distal to the MSS membrane 13. FIG. Therefore, it is presumed that the target preferentially binds to the aptamer on the distal end side from the MSS membrane 13 . In this case, even if steric hindrance due to the target occurs around the target-bound aptamer, other aptamers are present on the MSS membrane 13 side compared to the target-bound aptamer.
  • the surrounding aptamers are less susceptible to steric hindrance caused by the target. Therefore, even if the target binds to the aptamer, the surrounding aptamers are less likely to move due to steric hindrance. Therefore, when the aptamers are immobilized at different distances with respect to the MSS membrane 13, distortion of the MSS membrane may occur on the MSS membrane 13 due to the displacement of the surrounding aptamers. relatively low. On the other hand, when the aptamers are immobilized on the MSS membrane 13 at substantially the same distance, when the aptamers bind to the targets, the surrounding aptamers are affected by steric hindrance caused by the targets.
  • the positions of the surrounding aptamers will move, and the possibility that the MSS membrane will be distorted due to the movement of the positions of the surrounding aptamers is also relatively high. That is, when the aptamers are immobilized on the MSS membrane 13 at approximately the same distance, the binding of one aptamer to the target on the MSS membrane 13 also causes the position of the surrounding aptamers to move. The distortion of the MSS film 13 is amplified. Therefore, when the aptamer is immobilized on the MSS membrane 13 at substantially the same distance, that is, when the linker length is substantially constant, the sensitivity of the MSS membrane 13 is presumed to be improved. be.
  • cross-linking agent having an active ester at both ends of the molecule a cross-linking agent having an N-hydroxysuccinimide active ester and a haloacetyl-reactive group at both ends, a cross-linking agent having an N-hydroxysuccinimide active ester and a pyridyldithiol-reactive group at both ends, DCC, EDC, NHS, Sulfo-NHS, acetic anhydride and the like.
  • the linker is represented, for example, by the following formula (1).
  • M 1 represents an atom bonded to the silane coupling agent on the MSS film
  • L 1 represents a region (group) derived from the silane coupling agent
  • L 2 represents a cross-linking Represents the agent - derived region (group)
  • L2 is optional
  • M2 represents the atom that binds to the crosslinker or NH in the affinity tag.
  • NH represents an amine derived from an amino group of a silane coupling agent having an amino group.
  • L 1 is, for example, (M 1 )—Si(CH 3 ) 2-m (OR 4 ) m —R 1 —(NH) or (M 1 )—Si(CH 3 ) 2-m (OR 4 ) m —R 2 —NH—R 3 —(NH).
  • R 1 is a straight or branched alkyl group having 1 to 5 carbon atoms.
  • R 2 and R 3 are, for example, each independently a linear or branched alkyl group having 1 to 5 carbon atoms, and may be the same or different. Examples of the alkyl group include methyl group, ethyl group, propyl group, butyl group and pentyl group.
  • R4 is, for example, a hydrogen atom or a bond.
  • m is 1 or 2;
  • L 1 is represented by, for example, (M 1 )-Si(OR 4 ) 2 --(CH 2 ) 3 --(NH).
  • R4 is, for example, a hydrogen atom or a bond.
  • the length of the linker is, for example, the shortest molecular chain length (main chain length) between the functional group on the MSS membrane (for example, the oxygen atom of the silanol group on the silicon membrane) and the affinity tag such as avidin. can be represented.
  • the main chain length of the linker is 1 to 20, and since it can improve the sensitivity of MSS, it is preferably 1 to 15, 1 to 13, 3 to 13, 5 to 13, 1 to 11, 3 to 11, 1 ⁇ 10, 3-10, 1-8, 3-8, 1-5, 1-3, 1 or 2.
  • avidin-biotin binding is used, but the third example is not limited to this, and a linker is directly bound to the hydroxyl group or phosphate group of the aptamer. good too.
  • the aptamer can be immobilized on the MSS membrane 13 by amidating the 3' terminal phosphate group and reacting it with the linker.
  • the reaction can be carried out, for example, by coating the MSS film 13 with a solution containing a silane coupling agent having a methacrylic group.
  • a linker is formed between the main chain or side chain of the amino acid derivative and the main chain or side chain of the amino acid of the avidins.
  • a possible cross-linking agent is reacted to bind one end of the cross-linking agent to the amino acid derivative on the MSS membrane 13 .
  • the surface of the MSS membrane after treatment with the amino acid derivative is washed, and a solution containing a cross-linking agent is applied to the MSS membrane 13 to bind the amino acid derivative and the cross-linking agent.
  • the conditions for the cross-linking reaction can be appropriately determined according to, for example, the type of cross-linking agent.
  • the avidins are bound to the other end of the cross-linking agent. Specifically, after cross-linking, the surface of the MSS membrane is washed, and a solution containing avidins is applied to bind the other end of the cross-linking agent to the main chain or side chain of amino acids of avidins. Then, the biotin-bound aptamer is brought into contact with the MSS membrane 13 treated in this way, and the aptamer can be immobilized by binding the biotins and the avidins.
  • the silane coupling agent is represented by, for example, Y—Si(CH 3 ) 3-n (OR) n .
  • the silane coupling agent is a silane coupling agent having a methacryl group
  • examples of n, R, and Y are given below.
  • the "n" is 2 or 3.
  • examples of R include alkyl groups such as methyl group and ethyl group; acyl groups such as acetyl group and propyl group; and the like.
  • Y is a reactive functional group terminated with a methacryl group.
  • the silane coupling agent having a methacrylic group includes, for example, 3-(methacryloyloxy)propylmethyldimethoxysilane (eg, KBM-502 (manufactured by Shin-Etsu Silicone Co., Ltd.)), 3-(methacryloyloxy)propyltrimethoxysilane (eg, KBM-503 (manufactured by Shin-Etsu Silicone Co., Ltd.), GENIOSIL (registered trademark) GF31 (manufactured by Asahi Kasei Wacker Silicone Co., Ltd.)), 3-(methacryloyloxy) propylmethyldimethoxysilane (e.g., KBE-502 (manufactured by Shin-Etsu Silicone Co., Ltd.)), ( 3-methacryloyloxypropyl)triethoxysilane (for example, KBE-503 (manufactured by Shin-Etsu Silicone Co
  • the amino acid or amino acid derivative has, for example, a functional group capable of reacting with a methacryl group and a carboxyl group.
  • the functional group capable of reacting with the methacrylic group include a thiol group (--SH).
  • examples of the amino acid or amino acid derivative having a thiol group include cysteine; cysteine having an amino group modified such as N-acetylcysteine; and the like.
  • the cross-linking agent can be appropriately determined according to, for example, the functional group of the amino acid derivative subjected to cross-linking and the functional group of the amino acid of the avidins to be subjected to cross-linking.
  • the cross-linking agent can refer to the description of the cross-linking agent in the case of using the amino group of the main chain or side chain of the amino acid in the third example.
  • the cross-linking agent is the side chain thiol group of the amino acid in the third example. can be used.
  • the cross-linking agent can be used for cross-linking when utilizing the carboxyl group of the main chain or side chain of the amino acid in the third example.
  • the description of the agent can be used.
  • the cross-linking agent is preferably a cross-linking agent that does not substantially cause self-condensation because the length of the linker can be made constant.
  • the sensitivity of MSS can be improved by a mechanism similar to the mechanism described in the third example.
  • Specific examples of cross-linking agents that do not substantially cause self-condensation include cross-linking agents having N-hydroxysuccinimide active esters at both ends, cross-linking agents having imide ester reactive groups at both ends, and maleimide groups and N-hydroxysuccinimide active groups.
  • cross-linking agent having an ester at both ends of the molecule, cross-linking agent having an N-hydroxysuccinimide active ester and a haloacetyl reactive group at both ends, cross-linking agent having an N-hydroxysuccinimide active ester and a pyridyldithiol reactive group at both ends, DCC , EDC, NHS, Sulfo-NHS, acetic anhydride and the like.
  • the linker is represented, for example, by the following formula (2).
  • M 1 represents an atom bonded to the silane coupling agent on the MSS film
  • L 1 represents a region (group) derived from the silane coupling agent
  • A is an amino acid derivative.
  • L2 represents a crosslinker - derived region ( group), L2 may or may not be present
  • M2 represents an atom bonded to the crosslinker or NH in the affinity tag.
  • L 1 is, for example, (M 1 )—Si(OR 4 ) 2 —(CH 2 ) 3 —OC( ⁇ O)—C(CH 3 ) 2 —(A).
  • R4 is, for example, a hydrogen atom or a bond.
  • L2 is absent, for example.
  • the length of the linker can be represented, for example, by the shortest molecular chain length (main chain length) between the functional group on the MSS membrane (eg, silanol group on the silicon membrane) and the affinity tag such as avidin. can.
  • the main chain length of the linker is 1 to 20, and since it can improve the sensitivity of MSS, it is preferably 1 to 15, 1 to 13, 1 to 11, 1 to 10, 1 to 8, 1 to 5, 1 ⁇ 3, 1 or 2.
  • avidin-biotin binding is used, but the fourth example is not limited to this, and a linker is directly bound to the hydroxyl group or phosphate group of the aptamer. good too.
  • the aptamer can be immobilized on the MSS membrane 13 by amidating the 3' terminal phosphate group and reacting it with the linker.
  • the site for immobilizing the aptamer on the MSS membrane 13 is not particularly limited, and examples include the 3' end or 5' end.
  • FIG. 4 is a schematic diagram showing the MSS2 of the second embodiment.
  • (A) is a schematic plan view of the MSS2
  • (B) is a schematic cross-sectional view of (A) viewed from the II-II direction.
  • the MSS 2 of Embodiment 2 comprises a sensor substrate 10, electrodes 11, aluminum wires 121 (circuits), MSS films 13A (films), piezoresistive elements 14, and support regions 15.
  • the aluminum wire 12 is composed of a conductive layer 121 .
  • the MSS film 13A is composed of a support layer 131 and an amorphous layer 132, and the amorphous layer 132 is laminated on the support layer 131.
  • the support layer 131 is made of crystalline silicon, for example.
  • the amorphous layer 132 is composed of amorphous silicon (eg, amorphized silicon).
  • one surface of the film is composed of amorphous molecules, so that the contact area with other substances and the number of atoms that can be used for bonding are reduced compared to the case where the film is composed only of crystalline molecules. can be relatively increased. Therefore, by including the amorphous layer 132, the MSS film 13A can immobilize a larger amount of the binding substance on the surface of the MSS film 13A than the MSS film 13 does.
  • the amorphous layer 132 may contain other molecules in addition to amorphous silicon.
  • the other molecules include silicon carbide (silicon carbide), oxides of silicon carbide, and the like.
  • FIG. 5 is a schematic cross-sectional view showing an example of a method for manufacturing the MSS2.
  • MSS is prepared.
  • MSS commercially available MSS may be used, or self-prepared.
  • the MSS film 13 of the MSS is composed of a support layer 131 composed of crystalline silicon. Therefore, in the manufacturing method of Embodiment 2, the amorphous layer 132 is formed on the surface of the support layer 131 .
  • the support layer 131 is subjected to an amorphization treatment (amorphization treatment).
  • amorphization treatment amorphization treatment
  • the crystalline silicon on the surface of the support layer 131 is made amorphous by the amorphization treatment, and as shown in FIG. Therefore, an amorphous layer 132 made of amorphous silicon is formed.
  • the amorphization treatment may be any treatment as long as it causes a change in crystal structure.
  • Growth treatment for example, plasma CVD (Chemical Vapor Deposition), reactive ion etching (RIE), etc.
  • RIE reactive ion etching
  • the manufacturing method of Embodiment 2 can manufacture MSS2.
  • an amorphous layer 132 is formed on the surface of the MSS film 13A. Therefore, in MSS2, in the MSS film, the surface area of the MSS film that can be bound to the binding substance is increased, and the contact area with other substances and the number of atoms that can be used for binding are increased as compared with MSS1. Relatively increasing. Therefore, in the MSS2 of Embodiment 2, the immobilization efficiency of the binding substance can be improved.
  • the entire surface of one surface of the MSS film 13A is the amorphous layer 132, but the present invention is not limited to this, and part of one surface may be the amorphous layer 132. . Further, in the MSS2 of Embodiment 2, part or the entire surface of both surfaces of the MSS film 13A may be the amorphous layer 132. FIG. As a result, it is possible to prevent damage to the piezoresistive element 14 and the like during modification processing of the MSS film in the manufacturing method of the MSS2, which will be described later.
  • the amorphization treatment was performed as the modification treatment of the MSS film, but the present invention is not limited to this, and the amorphization treatment is combined with other modification treatments. good too. Another example of the method for manufacturing MSS will be described with reference to FIGS.
  • the silicon carbide oxide layer 135 on the surface contains silicon carbide oxide, the types of functional groups increase compared to crystalline silicon and amorphous silicon. It is excellent in reactivity with the above-mentioned cross-linking agents such as Specifically, since the MSS film 13B contains carbon atoms in its surface layer, carbon atoms can be selected as atoms to be bonded in addition to silicon atoms. Therefore, according to MSS2, binding molecules such as nucleic acid molecules or proteins can be introduced more easily. Therefore, according to MSS2, the binding substance can be more efficiently immobilized using the cross-linking agent.
  • FIG. 7 is a schematic diagram showing another example of a method for producing MSS2.
  • the MSS film 131 which is a crystalline silicon film, is subjected to the amorphization treatment to form an amorphous layer 132 (FIG. 7B).
  • carbon is introduced into the amorphous layer 132 to form a silicon carbide layer 133 containing silicon carbide (FIG. 7(C)), after which part of it is converted into an oxide layer 135 of silicon carbide. (Fig. 7(D)).
  • the crystalline silicon film 131 which is the MSS film, is subjected to an amorphization treatment by ion beam treatment. Thereby, an amorphous layer 132 is formed on the surface of the support layer 131 as shown in FIG. 7B.
  • the method for introducing the carbon can be implemented by, for example, a vapor deposition method, a CVD method, a coating method, or the like.
  • heat treatment may be performed in order to promote the reaction between the introduced carbon and the silicon in the amorphous silicon.
  • the lower limit of the temperature in the heat treatment is preferably 100° C. or higher, 200° C. or higher, or 300° C. or higher, for example.
  • the upper limit of the temperature in the heat treatment is, for example, 400° C. or less.
  • the temperature in the heat treatment is, for example, 100 to 400°C, 200 to 400°C, 300 to 400°C, and about 350°C.
  • the heat treatment time is, for example, 60 to 180 minutes.
  • the carbon layer 134 is removed from the MSS film after the introduction of carbon by plasma treatment in an oxygen atmosphere. Thereby, as shown in FIG. 7D, a silicon carbide layer 133 and a silicon carbide oxide layer 135 are formed.
  • oxygen plasma treatment for example, the above explanation of the oxygen plasma treatment can be used.
  • the removal of the carbon layer 134 may be performed by a method other than the oxygen plasma treatment, and specific examples thereof include sputtering treatment in an inert gas atmosphere such as helium, neon, argon, and dry etching treatment. .
  • Embodiment 2 can manufacture another example of MSS2.
  • FIG. 8 is a schematic diagram showing the MSS3 of the third embodiment.
  • the MSS 3 of Embodiment 3 comprises a sensor substrate 10, electrodes 11, aluminum wires 12A (circuits), MSS films 13B (films), piezoresistive elements 14, and support regions 15.
  • Aluminum wire 12A is composed of conductive layer 121 and insulating layer 122 .
  • the MSS film 13B is composed of a support layer 131 and an amorphous layer 132, and the amorphous layer 132 is composed of a silicon carbide layer 133 and a silicon carbide oxide layer 135.
  • the MSS film 13B is supported by the sensor substrate via a support region 15 having a piezoresistive element 14 formed thereon. Also, the MSS film 13B is connected to the aluminum wire 12A through the piezoresistive element 14, and the aluminum wire 12A is connected to the electrode 11, respectively.
  • the aluminum wire 12A constitutes a Wheatstone bridge circuit including four piezoresistive elements 14. As shown in FIG. Except for this point, the MSS3 of Embodiment 3 has the same configuration as the MSS1 of Embodiment 1, and the description thereof can be used. Although the MSS 3 of Embodiment 3 includes the electrode 11, the electrode 11 has any configuration and may or may not be present. If the MSS 2 does not have the electrode 11, the aluminum wire 12A is connected to the voltage applying device.
  • an insulating layer 122 is formed on the surface of the aluminum wire 12A, which is the line. Therefore, in the MSS3, since the insulation between the aluminum wires 12A is ensured, it is possible to perform measurement in a liquid phase such as a sample liquid. Therefore, MSS3 can be suitably used for the analysis of liquid samples.
  • an amorphous layer 132 is formed on the surface of the MSS film 13B, and the amorphous layer 132 is composed of a silicon carbide layer 133 and a silicon carbide oxide layer 135 . Further, in the MSS film 13B, an oxide layer 135 of silicon carbide is arranged (stacked) on the surface.
  • the silicon carbide oxide layer 135 on the surface contains silicon carbide oxide, the types of functional groups increase compared to crystalline silicon and amorphous silicon. It is excellent in reactivity with the above-mentioned cross-linking agents such as Therefore, according to MSS3, the binding substance can be more efficiently immobilized using the cross-linking agent.
  • MSS3 of Embodiment 3 was manufactured, and it was confirmed that a silicon carbide layer and a silicon carbide oxide layer were formed on the MSS film.
  • An insulating layer 122 was formed by oxidizing the aluminum wire 121 of the MSS. Specifically, nitric acid (1.38 g/ml or more) was applied onto the aluminum wire 121 of the MSS. The amount of nitric acid applied was 50 ⁇ l per MSS. After the coating, the aluminum wire 121 was oxidized by reacting at room temperature (about 25° C., hereinafter the same) for 60 minutes. After the oxidation treatment, the MSS was rinsed under pure water at room temperature for 30 minutes to remove nitric acid.
  • FIG. 9 is a photograph showing an electron microscope image.
  • the MSS film 13B consists of a support layer 131 made of crystalline silicon, a silicon carbide layer 133 made of amorphous silicon and silicon carbide, and a silicon carbide layer 133 made of amorphous silicon and silicon carbide. It was confirmed that an oxide layer 135 of silicon carbide composed of an oxide of silicon carbide was laminated. It was also found that when the MSS film was processed under the conditions described above, the thickness of the silicon carbide layer 133 was about 70 nm, and the thickness of the silicon carbide oxide layer 135 was about 5 nm. Amorphous silicon also has an improved surface area compared to crystalline silicon.
  • ⁇ Appendix> Some or all of the above-described embodiments and examples can be described as in the following appendices, but are not limited to the following.
  • ⁇ First Membrane Surface Stress Sensor> (Appendix 1) comprising a membrane and a sensor substrate;
  • the membrane is a membrane that deforms in response to surface stress, the sensor substrate comprises a support area and circuitry; the support region supports the membrane and comprises a piezoresistive element;
  • the piezoresistive element is an element that detects deformation of the film,
  • the circuit is connected to the piezoresistive element,
  • the circuit includes a metal capable of forming an oxide film by oxidation treatment,
  • the circuit has an oxide film layer on the surface, Membrane type surface stress sensor.
  • Appendix 2 2.
  • the membrane comprises a support layer; the support layer comprises a crystalline component of the amorphous layer; 6. The sensor of any one of Clauses 2-5, wherein the amorphous layer is laminated to the support layer.
  • the support layer comprises crystalline silicon; wherein the amorphous layer comprises amorphous silicon or amorphized silicon; A sensor according to Appendix 6.
  • the amorphous layer is a silicon carbide layer; an oxide layer of silicon carbide; the silicon carbide layer includes amorphous silicon or amorphized silicon and silicon carbide; the oxide layer comprises amorphous silicon or amorphized silicon and silicon carbide and/or oxidized silicon carbide; 8.
  • (Appendix 9) 9. The sensor of any one of the clauses 1-8, wherein the membrane is a silicon membrane.
  • (Appendix 10) 10. The sensor of any preceding clause, wherein the support region partially supports the membrane.
  • the support region includes a plurality of piezoresistive elements, 11. The sensor according to any one of appendices 1 to 10, wherein the circuit constitutes a Wheatstone bridge circuit including the plurality of piezoresistive elements.
  • (Appendix 12) the sensor substrate having a plurality of support areas; 12. The sensor of any one of Clauses 1 to 11, wherein the plurality of support regions each support the membrane.
  • (Appendix 13) 13 The sensor of any one of Clauses 1-12, wherein the metal is a valve metal.
  • Appendix 14 14. A sensor according to any preceding clause, wherein the metal is aluminium, tantalum, niobium, titanium, hafnium, zirconium, zinc, tungsten, bismuth, antimony, molybdenum, chromium, iron, or alloys thereof.
  • Appendix 15 15. The sensor of any one of Clauses 1-14, wherein the metal is aluminum.
  • the membrane comprises a binding substance that binds to a target; 16. The sensor of any one of Clauses 1 to 15, wherein the membrane deforms upon binding of a target to the binding substance. (Appendix 17) 17.
  • the sensor of clause 16, wherein the binding substance is a nucleic acid molecule or a protein.
  • Appendix 18 18.
  • the sensor of clause 16 or 17, wherein the binding substance is immobilized on one surface of the membrane.
  • Appendix 19 18.
  • the sensor of clause 16 or 17, wherein the binding substance is immobilized on both sides of the membrane.
  • Appendix 20 20.
  • Appendix 21 The membrane comprises a metal film on the surface, 21.
  • ⁇ Second Membrane Surface Stress Sensor> comprising a membrane and a sensor substrate;
  • the membrane is a membrane that deforms in response to surface stress, the sensor substrate comprises a support area; the support region supports the membrane and comprises a piezoresistive element;
  • the piezoresistive element is an element that detects deformation of the film, the film comprises an amorphous layer; Membrane type surface stress sensor.
  • Appendix 28 28.
  • the sensor of Clause 27, wherein the amorphous layer comprises amorphous silicon or amorphized silicon.
  • (Appendix 29) 29 The sensor of clause 27 or 28, wherein the amorphous layer comprises silicon carbide.
  • the amorphous layer is a silicon carbide layer; an oxide layer of silicon carbide; the silicon carbide layer includes amorphous silicon or amorphized silicon and silicon carbide; the oxide layer comprises amorphous silicon or amorphized silicon and silicon carbide and/or oxidized silicon carbide; 30.
  • the membrane comprises a support layer; the support layer comprises a crystalline component of the amorphous layer; 31.
  • the support layer comprises crystalline silicon; wherein the amorphous layer comprises amorphous silicon or amorphized silicon; 32.
  • the sensor of clause 31. (Appendix 33)
  • the amorphous layer is a silicon carbide layer; an oxide layer of silicon carbide; the silicon carbide layer includes amorphous silicon or amorphized silicon and silicon carbide; the oxide layer comprises amorphous silicon or amorphized silicon and silicon carbide and/or oxidized silicon carbide; 33.
  • (Appendix 34) 34 34.
  • the sensor substrate has a circuit, The support region includes a plurality of piezoresistive elements, 36.
  • Appendix 37 the sensor substrate having a plurality of support areas; 37.
  • the membrane comprises a binding substance that binds to a target; 38.
  • (Appendix 39) 39 The sensor of clause 38, wherein said binding agent is a nucleic acid molecule or a protein.
  • (Appendix 40) 40 The sensor of clause 38 or 39, wherein the binding substance is immobilized on one surface of the membrane.
  • (Appendix 41) 40 The sensor of clause 38 or 39, wherein the binding substance is immobilized on both sides of the membrane.
  • (Appendix 42) 42 The sensor according to any one of Appendices 38 to 41, wherein the binding substance is immobilized on the membrane via a conjugate of avidin or an avidin derivative and biotin or a biotin derivative.
  • the membrane comprises a metal film on the surface, 43.
  • Appendix 52 52.
  • Appendix 53 53.
  • Appendix 54 54.
  • (Appendix 62) 62 The manufacturing method according to appendix 61, wherein the amorphization treatment is performed by an amorphization treatment.
  • (Appendix 63) 63 The manufacturing method according to appendix 61 or 62, including the step of introducing carbon into the amorphous layer.
  • (Appendix 64) 64 The manufacturing method according to appendix 63, comprising removing carbon from the film by plasma treatment and/or sputtering treatment in an oxygen atmosphere.
  • (Appendix 65) 65 The manufacturing method according to any one of appendices 61 to 64, wherein the film is a silicon film.
  • ⁇ Method for manufacturing a semiconductor device> (Appendix 66) In a semiconductor device comprising a film and a circuit, a step of oxidizing a circuit containing a metal capable of forming an oxide film by oxidation treatment to form an oxide film layer on the surface of the circuit; A method of manufacturing a semiconductor device, comprising: subjecting the film to an amorphization treatment to form an amorphous layer on the surface of the film. (Appendix 67) 67. The manufacturing method according to appendix 66, wherein the oxidation treatment is performed by acid treatment and/or plasma treatment in an oxygen atmosphere. (Appendix 68) a step of acid-treating the circuit to form an oxide film layer on the surface of the circuit; 68.
  • the manufacturing method according to appendix 66 or 67 comprising performing a plasma treatment in an oxygen atmosphere on the circuit.
  • Appendix 69 69.
  • Appendix 70 69.
  • Appendix 71 71.
  • Appendix 72 72.
  • Appendix 77 77.
  • Appendix 78 78.
  • ⁇ Target analysis method> (Appendix 79) an applying step of applying a voltage to the membrane surface stress sensor in the sample liquid; an analysis step of analyzing the target in the sample liquid by measuring the stress change of the piezoresistive element in the film-type surface stress sensor; 48.
  • the present invention is useful in, for example, the analysis field of samples and the like, the medical field, and the like.

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Abstract

L'invention concerne un capteur de tension de surface de type film ayant une isolation entre des circuits.　Un capteur de tension de surface de type film 1 selon la présente invention comprend un film 13 et un substrat de capteur 10. Le film 13 se déforme en réponse à une tension de surface. Le substrat de capteur 10 comprend une région de support 15 et un circuit 12A. La région de support 15 supporte le film 13 et comprend un élément piézorésistif 14. L'élément piézorésistif 14 détecte la déformation du film 13. Le circuit 12A est relié à l'élément piézorésistif 14. Le circuit 12A comprend un métal qui peut être amené à former un film d'oxyde par un traitement d'oxydation. Le circuit 12A comprend une couche de film d'oxyde 122 sur sa surface.
PCT/JP2021/025694 2021-07-07 2021-07-07 Capteur de tension de surface de type film et procédé de fabrication de capteur de tension de surface de type film WO2023281674A1 (fr)

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PCT/JP2021/025694 WO2023281674A1 (fr) 2021-07-07 2021-07-07 Capteur de tension de surface de type film et procédé de fabrication de capteur de tension de surface de type film
JP2023532962A JPWO2023281674A1 (fr) 2021-07-07 2021-07-07

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PCT/JP2021/025694 WO2023281674A1 (fr) 2021-07-07 2021-07-07 Capteur de tension de surface de type film et procédé de fabrication de capteur de tension de surface de type film

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JP2016045032A (ja) * 2014-08-21 2016-04-04 日本電信電話株式会社 生体分子検出素子
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WO2020218086A1 (fr) * 2019-04-26 2020-10-29 国立研究開発法人物質・材料研究機構 Film sensible destiné à un capteur nanomécanique utilisant un poly(2,6-diphényl-p-phénylène oxyde), capteur nanomécanique comprenant ledit film sensible, procédé de revêtement de capteur nanomécanique avec ledit film sensible, et procédé de régénération de film sensible dudit capteur nanomécanique
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NISHIKAWA MICHIHIRO, MURATA TOMOHIRO, ISHIHARA SHINSUKE, SHIBA KOTA, SHRESTHA LOK KUMAR, YOSHIKAWA GENKI, MINAMI KOSUKE, ARIGA KAT: "Discrimination of Methanol from Ethanol in Gasoline Using a Membrane-type Surface Stress Sensor Coated with Copper(I) Complex", BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN, CHEMICAL SOCIETY OF JAPAN,NIPPON KAGAKUKAI, JP, vol. 94, no. 2, 15 February 2021 (2021-02-15), JP , pages 648 - 654, XP093022857, ISSN: 0009-2673, DOI: 10.1246/bcsj.20200347 *

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