KR101895123B1 - Double nano-pore device and method of manufacturing the same - Google Patents

Abstract

The dual nanopore element includes a spacer unit formed at a first height to accommodate a solution containing an object to be examined, a spacer unit disposed at an upper portion of the spacer unit, the spacer unit including a first reservoir and a second reservoir A first nanopore unit having a first nanopore capable of supplying the solution and a second nanopore unit disposed below the spacer unit and having a second reservoir and a second nanopore unit capable of discharging the solution from the accommodation space to the second reservoir, And a second nanopore unit having a pore.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a double-

The present invention relates to a dual nanopore device and a method of manufacturing the same, and more particularly, to a dual nanopore device capable of analyzing a nanoscale material such as DNA and a method of manufacturing the dual nanopore device.

Nanopore devices can control the passage of very small amounts of ions, and can also analyze materials passing through the nanopore. This is similar to the principle of a conventional Coulter counter, but it has recently been studied actively with respect to the nanopore device because it has a pore size of 1 nm or less and a fine nanopore of 100 nm or less.

For example, a nanopore device has a structure similar to an ion channel of a living organism. Therefore, the nanopore device is capable of passing through a nanopore formed in the nanopore device, such as A, G, T, C And the like. When a material such as the DNA passes through the nanopore, there is an energy barrier due to electrostatic interaction or geometric limitation. Therefore, the DNA can pass through the energy barrier. As a result, the DNA passing through the inside of the nanopore occupies the inside of the nanopore, so that the DNA blocks the flow of current due to the movement of ions present in the electrolyte. As a result, when there is nothing in the nanopore, a relatively low potential difference occurs when the DNA occupies the inside of the nanopore as compared with the potential at which only normal ions pass easily, ) Is reduced. Research on the sequencing of the DNA using the size of the blocking current is under way, which is called a blocking current.

In addition, research is underway to analyze nanomaterials that are smaller than nanopores, such as microRNAs, nanowires, and polymer chains, as well as DNA.

However, the pair of electrodes included in the nanopore device are located in a storage container in which an object such as the DNA generally located above and below the pore is stored. In this case, disturbances such as movement of the ion materials themselves or their ionic materials other than the subject existing between the two electrodes may occur. The accuracy of the inspection may deteriorate due to the influence of the disturbance.

Further, a part of the substance to be passed through the entrance or exit of the nanopore is adhered and cloaking occurs in the nanopore, so that the entrance of the nanopore is closed and slowly opened again so that the subject moves into the nanopore A phenomenon may occur. As a result, it is difficult to precisely confirm the movement of the object, and it is difficult to detect a signal due to the flow of the object within the nanopore. As described above, when a single nanopore is used, it is difficult to judge whether the nanomaterial has actually passed through with high reliability.

It is therefore an object of the present invention to provide a dual nanopore device capable of improving inspection accuracy and reliability using a double nanopore for a solution containing an object to be inspected will be.

Another object of the present invention is to provide a method for producing the above dual nanopore device.

A dual nano-pore device according to an embodiment of the present invention includes a spacer unit formed at a first height to accommodate a solution containing an object to be tested, a spacer unit disposed on the spacer unit, A first nanopore unit having a first nanopore unit capable of supplying the solution from the first reservoir to the reservoir space and a second nanopore unit disposed below the spacer unit, And a second nanopore unit having a second nanopore that can be discharged to the second nanopore unit.

In one embodiment of the present invention, the first and second nanopore units may be arranged asymmetrically about the spacer unit.

In one embodiment of the present invention, the first and second nanopores may not overlap each other when viewed in a plan view.

Here, a channel for stretching the test object may be formed in the receiving space between the first and second nanopores.

In addition, the length of the channel may be longer than the length of the subject.

In one embodiment of the present invention, the first height may be less than 5 times the length of the test object.

In an embodiment of the present invention, the first and second nanopores may have different diameters.

In one embodiment of the present invention, a first electrode, a reference electrode, and a second electrode, which can be electrically connected to the solution present in each of the first reservoir, the accommodation space, and the second reservoir, may be provided.

Here, the first and second electrodes and the reference electrode may have a wire shape.

In addition, the first and second electrodes and the reference electrode may include an Ag / AgCl alloy, graphene, or MoS2.

According to the method of manufacturing a dual nano-pore device according to an embodiment of the present invention, a spacer unit in which a space for accommodating a solution containing an object to be inspected is formed at a first height is prepared. On the top of the spacer unit, a first nanopore unit having a first reservoir and a first nanopore capable of supplying the solution from the first reservoir to the accommodating space is formed. On the other hand, a second nanopore unit having a second reservoir and a second nanopore that can be discharged to the second reservoir from the reservoir is formed in the lower portion of the spacer unit.

In one embodiment of the present invention, each of the first and second nanopore units may be attached to the upper and lower portions of the spacer unit using an adhesive member.

According to the dual nanopore device according to the embodiments of the present invention, a structure in which first and second nanopores and channels are structurally combined is formed. At this time, the first and second nanopores do not overlap each other. Therefore, a channel in which the moving direction of the subject moves to enter the second nanopore is formed inside the spacer unit, so that the shape of the subject can be adjusted or a signal of higher quality can be detected.

1 is a cross-sectional view illustrating a dual nanopore device according to an embodiment of the present invention.
2 is a cross-sectional view illustrating the operation of the dual nano-pore device of FIG.
3 is a flowchart illustrating a method of manufacturing a dual nanopore device according to an embodiment of the present invention.
4 is a cross-sectional view illustrating a method of fabricating a dual nano-pore device according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. The present invention is capable of various modifications and various forms, and specific embodiments are illustrated in the drawings and described in detail in the text. It should be understood, however, that the invention is not intended to be limited to the particular forms disclosed, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. In the accompanying drawings, the sizes and the quantities of objects are shown enlarged or reduced from the actual size for the sake of clarity of the present invention.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In the present application, the terms "comprise", "comprising", and the like are intended to specify that there is a feature, step, function, element, or combination of features disclosed in the specification, Quot; or " an " or < / RTI > combinations thereof.

On the other hand, unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries are to be interpreted as having a meaning consistent with the contextual meaning of the related art and are to be interpreted as either ideal or overly formal in the sense of the present application Do not.

Nanopore element

1 is a cross-sectional view illustrating a dual nanopore device according to an embodiment of the present invention. 2 is a cross-sectional view illustrating the operation of the dual nano-pore device of FIG.

Referring to FIGS. 1 and 2, a dual nanopore device 100 according to an embodiment of the present invention includes a spacer unit 110, a first nanopore unit 130, and a second nanopore unit 150 do.

The spacer unit 110 is formed with a receiving space 115 capable of receiving a solution containing the object. The receiving space 115 is formed at a first height h.

The solution may be temporarily stored in the accommodation space 115. Therefore, as the subject to be inspected temporarily remains in the accommodating space 115, the inspection time for the subject can be increased. In addition, the spacer unit 110 may support the first and second nanopores 130 and 150 on the upper and lower portions of the spacer unit 110, respectively.

On the other hand, the first height h of the receiving space 115 may correspond to the width of the path through which the test object can pass.

In one embodiment of the present invention, the first height h may be less than 5 times the length of the subject. Further, the first height h may be equal to or less than the length of the test object.

In one embodiment of the present invention, the spacer unit 110 may further include an adhesive member 111 (see FIG. 5) in which a through hole (corresponding to the accommodation space) is formed. Thus, the first and second nanopore units 130 and 150 may be bonded to the upper surface and the lower surface of the spacer unit 110, respectively.

Alternatively, the spacer unit 110 may correspond to a coating layer having a through-hole formed therein.

The spacer unit 110 may be formed of an insulating material as a whole. Thereby, the spacer unit 110 can electrically insulate the solution contained in the accommodation space 115 from the outside. The material forming the spacer unit 110 may be an insulating material or a dielectric material having an insulating property. For example, glass, mica, polymer, SiN, Al ₂ O ₃ , SiO ₂ , HfO ₂ , hBN and the like can be used.

Alternatively, the spacer unit 110 may be made of a polymer. In this case, the spacer unit 110 may have a film form or a coating layer form. The spacer unit 110 made of a polymer having a viscosity can be strongly bonded to each of the first and second nanopore units 130 and 150 without a separate adhesive.

The first nanopore unit 130 is disposed on the spacer unit 110.

The first nanopore unit 130 includes a first storage member 131 having a first reservoir 137 and a first membrane 133 having a first nanopore 135 formed therein.

 The solution containing the test object is supplied to the first reservoir 137 from above. The first reservoir 137 may be defined as a sidewall having a horizontal diameter that becomes smaller as it goes down. Thus, the first reservoir 137 may have a cone shape.

A first nanopore 135 is formed on the first membrane 133. The solution can pass from the first reservoir 137 to the accommodating space 115 through the first nanopore 135. At this time, if the test object remains in the first nanopore 135, a first blocking current may be measured between the first reservoir 137 and the accommodation space 115.

More specifically, when the solution containing no analyte passes through the first nanopore 135, there is no substance that partially blocks the first nanopore 135, so that a normal current is generated by the electrolyte in the solution Can be measured. On the other hand, when the solution in which the analyte dissolves passes through the first nanopore 135, the test object partially blocks the first nanopore 135, so that the electrical resistance may increase. As a result, the current flowing between the first reservoir (137) and the accommodation space (115) is reduced. At this time, the amount of current reduced based on the normal current is defined as the blocking current.

The average diameter of the object can be measured according to the blocking current value. For example, when the blocking current is relatively high, the subject has a relatively large diameter, whereas when the blocking current is relatively low, the subject has a relatively small diameter.

The first nanopore 135 may have a cylindrical shape. At this time, the first nanopore 135 may have a horizontal diameter of 1,000 nm or less and a vertical length of 10,000 nm or less. Alternatively, the first nanopore 135 may be in the form of an hourglass or a cone.

The second nanopore unit 150 is disposed below the spacer unit 110. The second nanopore unit 150 may be positioned asymmetrically with respect to the first nanopore unit 130 with respect to the spacer unit 110.

The second nanopore unit 150 has a second storage member 151 having a second reservoir 157 and a second membrane 153 having a second nanopore 155 formed therein.

In the second reservoir 157, the solution passing through the second nanopore 155 from the accommodation space 115 is stored. When the solution passes from the accommodation space 115 to the second nanopore 155, a second blocking current between the accommodation space 115 and the second reservoir 157 can be measured.

Accordingly, since the dual nanopore element 100 can measure both the first and second blocking currents, it is possible to secure an improved accuracy.

In an embodiment of the present invention, the first and second nanopores 135 and 155 may be positioned so as not to overlap each other when viewed in plan view. Thus, the solution containing the test object flows in the vertical direction through the first nanopore 135, horizontally moves in the accommodation space 115, and then flows through the second nanopore 155 2 < / RTI > As a result, the double-nanopore element 100 can have an improved accuracy because the object to be inspected can secure the residual time remaining in the accommodation space 1155.

2, a channel 117 (see FIG. 2) is formed in the accommodation space 115 in an accommodation space 115 capable of stretching the test object 10 between the first and second nanopores 135 and 155 May be formed. Thereby, when the subject 10 is moved through the channel 117, the subject 10 can more easily enter the second nanopore 155 in an unfolded state. As a result, closure of the second nanopore 155 can be inhibited from closing the entrance of the second nanopore 155.

For example, when the subject is DNA, the length is determined by the number of bases. Generally, when DNA moves in a solution in a narrow channel, the DNA is partially or wholly expanded in the flow direction of the fluid. Therefore, not only is it spread while passing through the first and second nanopores 135 and 155, Even when passing through the channel 117 formed in the unit 110, it is possible to relatively easily enter into the second nanopore 155 by stretching.

The horizontal length of the channel 117 formed between the first and second nanopores 135 and 155 may be longer than the length of the subject 10. Thereby, a sufficient time can be secured that the subject 10 passing through the channel 117 can be stretched.

Meanwhile, when the first and second nanopores 135 and 155 have a cylindrical shape, the first and second nanopores 135 and 155 may have different diameters. Accordingly, as the first and second nanopoles 135 and 155 are provided, the diameters of the first and second nanopores 135 and 155 are the same as the blocking current measured when passing through the first and second nanopores 135 and 155, . As a result, it is possible to more accurately examine the subject by using different signal values.

1 and 3, a first electrode 171 that can be electrically connected to a solution existing in the first reservoir 137, the accommodating space 115, and the second reservoir 157, (170) and a second electrode (172). The first blocking current flowing between the first electrode 171 and the reference electrode 170 and the second blocking current flowing between the second electrode 172 and the reference electrode 170 can be measured respectively, Examination of the carcass may be possible.

The first electrode 171 and the second electrode 172 may be inserted into the first and second reservoirs 137 and 157 so as to protrude from the inside of the first and second reservoirs 137 and 157, respectively. The first electrode 171 and the second electrode 172 may protrude from the first and second storage members 131 and 151, respectively. Meanwhile, the reference electrode 170 may be formed on the surface of the spacer unit 110 or inside thereof.

Here, the first and second electrodes 171 and 172 and the reference electrode 170 may have a wire shape. The first and second electrodes 171 and 172 and the reference electrode 170 may be formed of a metal such as gold, silver or copper or a metallic compound such as AgCl or a carbon nano material such as graphene or reduced graphene and it may include a two-dimensional plane and its composite materials such as a composite thereof, or _{MoS 2, WS 2, h-} BN.

Alternatively, the electrodes 170, 171, and 172 may include an electrochemically stable material whose electrochemical reaction is inhibited with respect to the analyte ions.

For example, the electrodes 170, 171, and 172 may be formed of a material having excellent electric conductivity such as silicon doped with a metal element. Furthermore, a modified two-dimensional planar material doped with an element such as boron (B) or nitrogen may be used for the two-dimensional planar material, and the element that can be doped is not particularly limited. In addition, as a composite of nanomaterials, electrodes such as graphene coated with gold nanoparticles or electrodes composed of a double layer of Cr / Au can be used.

Manufacturing method of nanopore device

3 is a flowchart illustrating a method of manufacturing a dual nanopore device according to an embodiment of the present invention. 4 is a cross-sectional view illustrating a method of fabricating a dual nano-pore device according to an embodiment of the present invention.

Referring to FIG. 3, in accordance with a method of manufacturing a dual nano-pore device according to an embodiment of the present invention, a spacer unit having a receiving space capable of accommodating a solution containing an object to be tested is formed at a first height (S110) . Subsequently, a first nanopore unit having a first reservoir and a first nanopore capable of supplying the solution from the first reservoir to the accommodating space is formed on the spacer unit (S120). On the other hand, a second nanopore unit having a second reservoir and a second nanopore that can be discharged to the second reservoir from the reservoir is formed in the lower portion of the spacer unit (S130). Thereby, a double nanopore element including the first and second nanopore units is manufactured.

Referring to FIGS. 3 and 4, each of the first and second nanopore units 130 and 150 may be formed through a photolithography process or an imprint process. For example, a silicon nitride (SiN) layer is grown on a silicon substrate. Thereafter, the silicon substrate is patterned using a photolithography process to form a first storage member 131 having a first reservoir. Thereafter, the thickness of the silicon substrate is adjusted through the backside etching process on the rear surface of the silicon substrate.

A first membrane 133 having a first nanopore may be formed using a beam such as a scanning electron microscope, a focused ion beam, or a transmission electron microscope to form a nanopore in the silicon nitride (SiN) layer. It can also produce pores with nanopatterning techniques such as E-beam lithography and nanoimprint or electrochemical etching techniques.

On the other hand, the second nanopore unit may also be formed through the same process as the first nanopore unit.

Next, the first and second nanopore units 130 and 150 may be coupled to the spacer unit 110. At this time, an adhesive is preferably coated on the upper and lower surfaces of the spacer unit 110 so that each of the first and second nanopore units 130 and 150 can be attached thereto.

At this time, when the first and second nanopore units 130 and 150 are attached to the spacer unit 110, the first and second nanopores 130 and 150 may be arranged so as not to overlap each other. have.

In one embodiment of the present invention, each of the first and second nanopore units 130 and 150 is attached to the top and bottom surfaces of the spacer unit 110, and then the silicon nitride layers are deposited on each of the silicon nitride layers It is possible to form each of the nanopores.

The double nanopore device can analyze an object that passes through various measurement signals more effectively. Therefore, the double nanopore device can be utilized not only for DNA and microRNA sequencing but also for analyzing various nanomaterials.

100: double nanopore element 110: spacer unit
130: first nanopore unit 150: second nanopore unit
170: reference electrode 171: first electrode
172: second electrode

Claims (12)

A spacer unit formed at a first height to accommodate a solution containing the object to be inspected;
A first nanopore unit disposed on the spacer unit and having a first reservoir and a first nanopore capable of supplying the solution from the first reservoir to the accommodating space; And
And a second nanopore unit disposed below the spacer unit, the second nanopore unit having a second reservoir and a second nanopore that can discharge the solution from the accommodation space to the second reservoir,
The first and second nanopores do not overlap each other when viewed in a plan view,
Wherein a channel for stretching the test object is formed between the first and second nanopores in the receiving space. The dual nanopore element according to claim 1, wherein the first and second nanopore units are arranged asymmetrically with respect to the spacer unit. delete delete The dual nanopore element according to claim 1, wherein the length of the channel is longer than the length of the test object. The dual nanopore element according to claim 1, wherein the first height is 5 times or less the length of the test object. The dual nanopore device according to claim 1, wherein the first and second nanopores have different diameters. The dual nano-pore device according to claim 1, comprising a first electrode, a reference electrode, and a second electrode that can be electrically connected to a solution present in each of the first reservoir, the accommodation space and the second reservoir . The dual nanopore element according to claim 8, wherein the first and second electrodes and the reference electrode have a wire shape. The dual nanopore element according to claim 8, wherein the first and second electrodes and the reference electrode comprise an Ag / AgCl alloy, graphene or MoS ₂ . Preparing a spacer unit in which a receiving space capable of receiving a solution containing an object to be inspected is formed at a first height;
Forming a first nanopore unit having a first reservoir and a first nanopore capable of supplying the solution from the first reservoir to the accommodation space, on top of the spacer unit; And
Forming a second reservoir and a second nanopore unit having a second nanopore that can be discharged to the second reservoir from the reservoir, at a lower portion of the spacer unit,
The first and second nanopores do not overlap each other when viewed in a plan view,
Wherein a channel capable of stretching the test object is formed between the first and second nanopores in the receiving space. 12. The method of claim 11, wherein each of the first and second nanopore units is attached to an upper portion and a lower portion of the spacer unit using an adhesive member.

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