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WO2016099108A1 - Élément nanopore et son procédé de fabrication - Google Patents

Élément nanopore et son procédé de fabrication Download PDF

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Publication number
WO2016099108A1
WO2016099108A1 PCT/KR2015/013705 KR2015013705W WO2016099108A1 WO 2016099108 A1 WO2016099108 A1 WO 2016099108A1 KR 2015013705 W KR2015013705 W KR 2015013705W WO 2016099108 A1 WO2016099108 A1 WO 2016099108A1
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Prior art keywords
pore
nano
layer
electrode
electrodes
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Ceased
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English (en)
Korean (ko)
Inventor
한창수
최욱
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Korea University Research and Business Foundation
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Korea University Research and Business Foundation
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/48707—Physical analysis of biological material of liquid biological material by electrical means
    • G01N33/48721—Investigating individual macromolecules, e.g. by translocation through nanopores
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles

Definitions

  • the present invention relates to a nano-pore device and a method for manufacturing the same, and more particularly, to a nano-pore device and a method for manufacturing the nano-pore device capable of analyzing nano-sized material such as DNA.
  • Nanopore devices have the ability to control very small amounts of ion transport and analyze materials passing through nanopores. Therefore, recently, the nano pore device has been actively studied.
  • the nanopore device has a structure similar to the ion channel of life, bases such as A, G, T, and C included in materials such as DNA passing through the nanopores formed in the nanopore device Sequencing is ongoing.
  • a material such as DNA passes through the nanopores
  • the DNA can pass through the energy barrier.
  • the DNA passing through the nanopores may block the flow of current due to the movement of ions flowing along the inside of the pore.
  • a relatively low potential difference occurs when the DNA passes as compared with the potential difference when only normal ions pass, thereby reducing current. This is called blocking current, and studies are underway to sequencing the DNA using the magnitude of the blocking current.
  • the pair of electrodes included in the nano-pore device is generally located in a storage container in which an object such as DNA is stored.
  • disturbance such as movement of ions or other materials except for the test object existing between the two electrodes occurs. Due to the disturbance may cause a problem that the accuracy of the inspection is deteriorated.
  • cloaking may occur at the inlet or the outlet of the nanopores, thereby closing the inlet of the nanopores and slowly opening them again, thereby moving the inspected object into the nanopores.
  • the thickness of the nanopore is too thick to analyze the nucleotide sequence, such as conventional DNA, it is very difficult to accurately measure the change in the blocking current according to the change in the base interval.
  • One object of the present invention is to provide a nano-pore device capable of accurately measuring the blocking current (blocking current) generated between the inlet and outlet of the nano-pores.
  • One object of the present invention is to provide a method for manufacturing the nano-pore device.
  • the nano-pore device is provided with a nano-pore structure formed with a nano-pore having an inlet and outlet so that the subject can pass through and spaced apart from each other between the inlet and the outlet, the end is inside the nanopore It is exposed to, and comprises a pair of first and second electrodes provided to measure the current flowing into the nano-pores.
  • the nano-pores have a size of less than 100 nm, may be provided to have a length of less than 10,000 nm.
  • each of the first and second electrodes is a metal such as gold, silver, palladium platinum, hafnium, copper and graphene, graphite, reduced graphene, h-BN, WS 2 , And two-dimensional materials such as MOS 2 .
  • each of the first and second electrodes may have a donut shape to penetrate the nano-pore structure in the horizontal direction.
  • each of the first and second electrodes are disposed to be spaced apart from each other by an insulating film formed in the vertical direction of the nano-pores, it may be formed so as not to overlap each other when viewed in plan view.
  • each of the first and second electrodes are disposed to be spaced apart from each other by an insulating film formed in the vertical direction of the nano-pores, it may be formed to face each other with respect to the nano-pores.
  • the insulating layer may be provided to have a tunneling current between the first and second electrodes by having a gap of 5 nm or less between the first and second electrodes.
  • the nano pore device may further include a third electrode disposed between the first and second electrodes and controlling a flow of the inspected object flowing inside the nano pore.
  • the third electrode may have an exposed portion exposed inside the nanopores at one end, and the exposed portion may be coated with a dielectric material.
  • a nano-pore device In the method of manufacturing a nano-pore device according to an embodiment of the present invention, after forming a nano-pore structure formed through the nano-pores penetrating the inlet and outlet so that the subject can pass, spaced apart between the inlet and outlet And a pair of end portions are exposed to the inside of the nano-pores and apply a voltage to the inside of the nano-pores to form a pair of first electrodes and second electrodes.
  • a pore layer is formed on a substrate, and a first electrode layer of a planar structure is formed on the pore layer to form the pore layer and the first electrode layer.
  • a first preliminary pore structure is formed.
  • a second electrode layer formed of a planar structure is formed on the interlayer insulating film.
  • an insulating film is formed on the second electrode layer, and the insulating film, the second electrode layer, the interlayer insulating film, the first electrode layer, and the pore layer are sequentially etched to have an inlet and an outlet so that the object can pass.
  • a nano pore structure, a first electrode and a second electrode, on which the nano pores are formed, are formed.
  • the step of partially etching the substrate to form a first opening at a position corresponding to the nano-pores may be additionally performed.
  • a storage container may be additionally attached so that the inlet is exposed on the insulating film pattern.
  • a) to form a first pore layer on a first substrate b) to form a first electrode layer of a planar structure on the first pore layer
  • a first preliminary pore structure including the first pore layer and the first electrode layer is formed.
  • c) forming an interlayer insulating film on the first electrode layer and d) preparing a second preliminary pore structure in which a second substrate, a second pore layer, and a second electrode layer are sequentially formed using steps a) to b).
  • step f) may include performing one of an anodical bonding process, a plasma bonding process or a microwave bonding process.
  • the first substrate before forming the interlayer insulating layer on the first electrode layer, the first substrate may be partially etched to form a first opening at a position corresponding to the nanopores.
  • the second substrate After forming the second electrode layer, the second substrate may be partially etched to form a second opening at a position corresponding to the nanopores.
  • the nano-pore device of the present invention at least two electrodes formed to be spaced apart from each other between the inlet and the outlet of the nano-pores.
  • the nano pore device can be utilized as a device for analyzing various nanomaterials as well as sequencing of DNA and micro RNA.
  • FIG. 1 is a cross-sectional view for explaining a nano-pore device according to an embodiment of the present invention.
  • FIGS. 2 and 3 are cross-sectional views and a plan view for explaining a nano-pore device according to an embodiment of the present invention.
  • FIG. 4 is a cross-sectional view illustrating a nano pore device according to an embodiment of the present invention.
  • 5 to 11 are cross-sectional views illustrating a method of manufacturing a nano-pore device according to the present invention.
  • FIGS. 12 to 14 are cross-sectional views illustrating a method of manufacturing a nano-pore device according to the present invention.
  • first and second may be used to describe various components, but the components should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from another.
  • the first component may be referred to as the second component, and similarly, the second component may also be referred to as the first component.
  • the interlayer distances are defined as the gaps between them that correspond to channels and do not include the thickness of a single layer.
  • the thickness of some single layer may be included in the interlayer spacing.
  • the nano-pore device is provided with a nano-pore structure formed with a nano-pore having an inlet and outlet so that the subject can pass through and spaced apart from each other between the inlet and the outlet, the end is inside the nanopore It is exposed to, and comprises a pair of first and second electrodes provided to measure the current flowing into the nano-pores.
  • FIG. 1 is a cross-sectional view for explaining a nano-pore device according to an embodiment of the present invention.
  • the nano pore device 10 includes a nano pore structure 100, a first electrode 210, and a second electrode 220.
  • the nano-pore structure 100 is formed with a nano-pore 105 having an inlet 101 and the outlet 103 so that the subject can pass.
  • the nano pores 105 may have a size of 100 nm or less, and may be provided to have a length of 10,000 nm or less. In addition, the nano-pores 105 may have a size that is wider or narrower toward the bottom.
  • the first electrode 210 and the second electrode 220 are spaced apart from each other between the inlet and the outlet. End portions of each of the first electrode 210 and the second electrode 220 are provided to be exposed into the nanopores 105. A voltage is applied into the nanopores 105 through the first electrode 210 and the second electrode 220, and the current flowing into the nanopores 105 may be measured. The exposed portions of the first electrode 210 and the second electrode 220 may be coated using a dielectric material.
  • the first electrode 210 and the second electrode 220 may be disposed in close proximity to ensure a constant resolution. However, the first electrode 210 and the second electrode 220 should be spaced apart so as not to influence each other electrically. That is, it is preferable that the separation distance is secured such that leakage current generated between the first and second electrodes 210 and 220 is smaller than noise generated by the flow of ions in the nanopore device.
  • the nano-pores 10 may be used.
  • the included first electrode 210 and the second electrode 220 are disposed between the inlet 101 and the outlet 103 defining the nanopores 105. Therefore, the influence of the disturbance occurring in the reservoir (reservoir) that is outside the nano-pores 105 and accommodates the test subject can be suppressed.
  • the nano pore device 10 may measure the change in electric field generated inside the nano pore 105 with improved sensitivity.
  • the pair of first and second electrodes 210 and 220 mounted inside the nanopores 105 are spaced at regular intervals, thereby interposing the first and second electrodes 210 and 220.
  • the blocking current can be precisely measured by the moving test object material.
  • each of the first and second electrodes 210 and 220 may have a thickness of 1 ⁇ m or less. However, in consideration of the size of the nano-pores 105, it is sufficient that the first and second electrodes 210 and 220 have a thickness that can be spaced apart from each other. There is no particular limitation on the thickness.
  • each of the first and second electrodes 210, 220 is a metal such as gold, silver, copper or a metal compound such as AgCl or graphene, carbon nano such as reduced graphene Materials and their complexes or two-dimensional planar materials such as MoS 2 , WS 2 , h-BN and their complexes can be used.
  • each of the first and second electrodes 210 and 220 may include an electrochemically stable material in which an electrochemical reaction is suppressed with respect to an object under test.
  • a material having a good electrical conductivity such as silicon doped with a metal element
  • a modified two-dimensional planar material doped with elements such as boron (B) and nitrogen (nitrogen) may be used as the two-dimensional planar material, and the dopable elements are not particularly limited.
  • an electrode as a composite may be used, such as a graphene coated with gold nanoparticles or a double layer of Cr / Au.
  • each of the first and second electrodes 210 and 220 may have a donut shape to penetrate the nano-pore structure 100 in the horizontal direction.
  • the first and second electrodes 210 and 220 may have a structure that is entirely enclosed along the circumferential direction of the nanopores 105.
  • the nano pore device 10 may measure the change in the electric field generated inside the nano pore 10 with improved sensitivity.
  • each end of the first and second electrodes 210 and 220 inserted into the nano pore structure 100 may be exposed into the nano pore 105. . Accordingly, each end of the first and second electrodes 210 and 220 may be in direct physical contact with the flowing ionic fluid. However, exposed exposed portions of the first and second electrodes 210 and 220 may be shielded by a dielectric material.
  • FIGS. 2 and 3 are cross-sectional views and a plan view for explaining a nano-pore device according to an embodiment of the present invention.
  • a nano pore device 10 includes a nano pore structure 100, a first electrode 210, and a second electrode 220.
  • the first electrode 210 and the second electrode 220 may be located close enough to measure the tunneling current (tunneling current).
  • the first electrode 210 and the second electrode 220 may each have a half donut shape.
  • An insulating film may be interposed between the first and second electrodes 210 and 220.
  • the first and second electrodes 210 and 220 may be symmetric about the nanopores 105.
  • a tunneling current may occur between the first and second electrodes.
  • the size of the tunneling current is changed to change between minute electrodes.
  • a tunneling current value between the first and second electrodes 210 and 220 when any one of the bases of DNA is located at the first and second electrodes 210 and 220. This can be changed.
  • the first and second electrodes 210 and 220 may be disposed not only to be spaced apart by the insulating layer but to overlap each other when viewed in plan view. That is, the first and second electrodes 210 and 220 are positioned to surround the nano pores 105 with respect to the nano pores 105 and are disposed so as not to overlap with each other when viewed in plan view. As a result, generation of a leakage current due to mutual interference between the first and second electrodes 210 and 220 may be suppressed.
  • the first and second electrodes 210 and 220 are disposed to face each other with respect to the nano pore 105.
  • the flow of the nanomaterial may interfere with the current flow in the horizontal direction between the first and second electrodes 210, 220. Can be. As a result, a blocking current value between the first and second electrodes 210 and 220 may be measured.
  • the first and second electrodes 210 and 220 may have a height difference in a forming position along the vertical direction in the nanopores 105. Therefore, as the size of the nano pores 105 is smaller, the current flow between the first and second electrodes 210 and 220 may be more sensitively disturbed.
  • the first and second electrodes 210 and 220 may be formed on the same plane, but such a method is very difficult to manufacture. Therefore, the method of manufacturing a nano-pore device having the above structure may be relatively easy.
  • FIG. 4 is a cross-sectional view illustrating a nano pore device according to an embodiment of the present invention.
  • a nano pore device includes a nano pore structure 100, a first electrode 210, and a second electrode 220. Furthermore, the nano pore device further includes a third electrode 230 between the first and second electrodes.
  • the voltage applied to the third electrode 230 is a gate voltage (V G ), the voltage applied to the first electrode 210 is a source voltage (V S ), the voltage applied to the second electrode 220 is It can be defined as the drain voltage (V D ).
  • An electrical signal unit (not shown) for applying an electrical signal to the first to third electrodes 210, 220, and 230 is provided.
  • the electrical signal unit may be connected to the first to third electrodes through a probe (not shown).
  • the nano-pore element 10 functions as an ionic field effect transistor (IFE) by the gate voltage V G , the source voltage V S , and the drain voltage V D applied by the electrical signal unit. can do.
  • IFE ionic field effect transistor
  • the ions contained in the test object moves through the nano-pores, ion current flows and the nano-pores 105 serve as channels for the movement of the ions.
  • the cations and anions ionized from the test subject may move in a specific direction by the source voltage V S and the drain voltage V D applied to the first and second electrodes 210 and 220, respectively. In this case, the on state and the off state of the transistor may be controlled by the gate voltage V G applied to the third electrode 230.
  • each end of the third electrode 230 embedded in the nano-pore structure 100 may be exposed into the nano-pores 105.
  • the end of the third electrode 230 may be in direct physical contact with the flowing ionic fluid.
  • the exposed exposed portion of the third electrode 230 may be coated with a dielectric material.
  • 5 to 11 are cross-sectional views illustrating a method of manufacturing a nano-pore device according to the present invention.
  • the pore layer 113 is formed on the substrate 110.
  • the pore layer nitrides the substrate 110 made of silicon.
  • the upper surface of the substrate 110 may be converted into silicon nitride to form a pore layer.
  • the substrate 110 is partially etched to form a first opening 111 at a position corresponding to the nanopores.
  • an etching process may be performed on the substrate 110 using the mask pattern as an etching mask. have.
  • a first preliminary pore structure including the pore layer 113 and the first electrode layer 115 is formed by forming a first electrode layer 115 having a planar structure on the pore layer 113. 117).
  • the first electrode layer is a metal such as gold, silver, copper or a metal compound such as AgCl or graphene, carbon nano material such as reduced graphene and a composite thereof or a two-dimensional plane such as MoS 2 , WS 2 , h-BN It can be formed using materials and their complexes.
  • the first electrode layer may include an electrochemically stable material that is suppressed electrochemical reaction to the subject ions.
  • a graphene thin film formed on a copper foil may be transferred onto the pore layer.
  • the graphene thin film may be formed through a chemical vapor deposition process.
  • an interlayer insulating layer 120 is formed on the first electrode layer 115.
  • the interlayer insulating layer 120 may be formed using a metal oxide such as aluminum oxide.
  • the interlayer insulating layer 120 may be formed through a chemical vapor deposition process.
  • a second electrode layer 125 having a planar structure is formed on the interlayer insulating layer 120.
  • the second electrode layer 125 is a metal such as gold, silver, copper or a metal compound such as AgCl or graphene, carbon nano material such as reduced graphene and a composite thereof or MoS 2 , WS 2 , h-BN It can be formed using a two-dimensional planar material and composites thereof.
  • the second electrode layer 125 may include an electrochemically stable material to suppress the electrochemical reaction to the subject ions.
  • a graphene thin film formed on a copper foil may be transferred onto the pore layer.
  • the graphene thin film may be formed through a chemical vapor deposition process.
  • each of the first and second electrode layers 115 and 125 may be patterned to have a predetermined shape (see FIGS. 2 and 3).
  • a photolithography process an electron beam lithography process, a focused ion beam (FIB) process, or a nanoimprinting process is used.
  • FIB focused ion beam
  • each of the first and second electrodes may be formed.
  • an insulating layer 140 is formed on the second electrode layer 125.
  • the insulating layer 140 may be formed using a metal oxide such as aluminum oxide.
  • the insulating layer 140 may be formed through a chemical vapor deposition process.
  • the insulating film 140, the second electrode layer 125, the interlayer insulating film 120, the first electrode layer 115, and the pore layer 113 are sequentially etched to pass the test object.
  • the nano-pore structure 100 on which the nano-pores 105 are formed includes a pore layer pattern 113a, an interlayer insulating layer pattern 120a, and an insulating layer pattern 140a.
  • the first electrode 210 and the second electrode 220 patterned from the first electrode layer 115 and the second electrode layer 125 may be formed.
  • a photolithography process, an electron beam lithography process, a focused ion beam process, or the like may be used to form the nanopores 105.
  • the nano-pores Compared to the case where the first and second electrodes are disposed in a reservoir containing a nano-pore element 10 according to embodiments of the present invention, which is located outside the nano-pores and accommodates the object under test, the nano-pores The first electrode 210 and the second electrode 220 included in the device 10 are disposed between the inlet 101 and the outlet 103 defining the nanopores 105. Therefore, the influence of the disturbance occurring in the receptacle that is present outside the nanopores 105 and accommodates the test object can be suppressed. Furthermore, the nano pore device 10 may measure the change in electric field generated inside the nano pore 105 with improved sensitivity.
  • the pair of first and second electrodes 210 and 220 mounted inside the nanopores 105 are spaced at regular intervals, thereby interposing the first and second electrodes 210 and 220.
  • the blocking current can be precisely measured by the moving test object material.
  • the first storage container 310 may be formed on the insulating layer pattern 140a to expose the inlet 101.
  • a second storage container 320 may be formed under the substrate to expose the outlet 103.
  • FIGS. 12 to 14 are cross-sectional views illustrating a method of manufacturing a nano-pore device according to the present invention.
  • a) a first pore layer 113 is formed on the first substrate 110, and a first electrode layer 115 having a planar structure is formed on the first pore layer 113.
  • a first preliminary pore structure 117 including the first pore layer 113 and the first electrode layer 115 is formed.
  • an interlayer insulating layer 120 is formed on the first electrode layer 115.
  • the second preliminary pore structure 417 in which the second pore layer 413 and the second electrode layer 415 are sequentially formed on the second substrate 410 is prepared by performing steps a) to b). .
  • the second preliminary pore structure 417 is positioned on the interlayer insulating layer 120 to face the first preliminary pore structure 117.
  • the interlayer insulating layer 210 and the second electrode layer 415 are attached.
  • the interlayer insulating layer 210 and the second electrode layer 415 may be attached by performing one of an anodical bonding process, a plasma bonding process, or a microwave bonding process.
  • an anodical bonding process e.g., a plasma bonding process
  • a microwave bonding process e.g., a microwave bonding process
  • the second pore layer 413, the second electrode layer 415, the interlayer insulating layer 210, the first electrode layer 115, and the first pore layer 113 are sequentially etched.
  • the nano-pore structure 100, the first electrode 210, and the second electrode 220, on which the nano-pores 105 having the inlets 101 and the outlets 103 are formed, may pass through the inspected object.
  • Nanopore devices can be applied to devices that analyze DNA while passing nanomaterials smaller than the size of nanopores such as microRNA, nanowires, and nanoscale polymers.

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Abstract

L'invention concerne un élément nanopore qui comprend : une structure de nanopore ayant un nanopore ayant un trou d'entrée et un trou de sortie, à travers lesquels un objet testé passe; et une paire de premières électrodes et de secondes électrodes, qui sont placées séparément entre le trou d'entrée et le trou de sortie, et dont une partie extrémité est exposée à l'intérieur du nanopore, de façon à mesurer un courant circulant à travers l'intérieur du nanopore.
PCT/KR2015/013705 2014-12-18 2015-12-15 Élément nanopore et son procédé de fabrication Ceased WO2016099108A1 (fr)

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KR10-2014-0183043 2014-12-18
KR1020140183043A KR101666725B1 (ko) 2014-12-18 2014-12-18 나노 포어 소자 및 이의 제조 방법

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CN108982612A (zh) * 2018-07-02 2018-12-11 浙江大学 基于纳米通道阵列表面喷镀金的集成式电化学电极系统
CN108982615A (zh) * 2018-07-02 2018-12-11 浙江大学 基于纳米通道阵列表面喷镀金/铂的集成式电化学电极系统
WO2020160559A1 (fr) * 2019-02-01 2020-08-06 Northeastern University Séquenceur de nanopores mxene de biopolymères
IT202100015821A1 (it) * 2021-06-17 2022-12-17 Elements S R L Dispositivo di rilevazione di nano particelle in un fluido.

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KR102176130B1 (ko) * 2018-11-13 2020-11-10 고려대학교 산학협력단 마이크로 포어를 이용한 생체 분자 검출 장치
KR102327635B1 (ko) * 2019-12-12 2021-11-17 광운대학교 산학협력단 나노 포어 필터 제조방법 및 이로부터 제조된 나노 포어 필터
CN112114020A (zh) * 2020-09-14 2020-12-22 安徽师范大学 双敏感金修饰的dna功能化玻璃纳米孔门控系统、构筑方法及应用

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