WO2019109254A1

WO2019109254A1 - Preparation method for and uses of nanopore and array

Info

Publication number: WO2019109254A1
Application number: PCT/CN2017/114643
Authority: WO
Inventors: 刘泽文; 王一凡; 陈琦
Original assignee: 清华大学
Priority date: 2017-12-05
Filing date: 2017-12-05
Publication date: 2019-06-13

Abstract

A preparation method for and uses of a nanopore. In the nanopore, a nanopore structure is implemented on one surface of a substrate by means of etching, and the other surface of the substrate is etched to reduce the thickness. When an opening event of a reference pore occurs due to the thickness reduction, a tracer gas (10) is packaged in a cavity. Atomic layer etching is carried on the other surface of the substrate, and the existence of the tracer gas (10) is detected at the same time; and when atoms of the tracer gas (10) are detected, it indicates that the opening event occurs, and the etching is stopped, so as to obtain a nanopore.

Description

Preparation method and use of nanopore and array

Technical field

The invention relates to the field of silicon-based three-dimensional processing technology, in particular to a preparation method and application of a nanopore and an array, in particular to a controllable silicon-based nanopore suitable for a biomacromolecule detection platform, a near-field optical and an atomic lithography grating. Production and feedback methods.

Background technique

Biomacromolecule sequencing, especially DNA single-base sequencing and protein sequencing, is one of the core technologies of modern life science research technology. Using solid-state nanopores as the basis for sequencing sensing, a low-cost, high-throughput, direct-reading detection platform is recognized as one of the most promising technologies.

The pore size of the solid nanopores can be flexibly controlled by humans, and has ideal biochemical pore stability, excellent physical and chemical properties. With the help of microscope electron beam technology, focused ion beam technology (FIB) and other processes, researchers have successfully produced a variety of nanometer, sub-nanometer solid pores with controlled pore size. However, the fabrication of solid-state nanopores also faces some problems. Firstly, the solid-state nanopore shape made by FIB has poor controllability and the channel is usually cylindrical. The effective length depends on the thickness of the film. The robustness and spatial resolution cannot be optimized at the same time. Secondly, all nanopore-based sequencing methods are used. There is currently no effective method for controlling the rate at which biomolecules pass through the nanopore, due to the too fast perforation rate and low temporal resolution.

Therefore, the current method for preparing nanopores still needs to be improved.

Summary of the invention

The present invention aims to solve at least one of the technical problems in the related art to some extent. It is an object of the present invention to provide a method for preparing nanopores which can achieve sub-nanometer precision and precise precision controllability.

To achieve the above objective, the technical solution of the present invention is implemented by the following steps:

According to a first aspect of the present invention, there is provided a method for preparing a nanopore comprising the steps of: (1) forming a first masking layer and a second masking layer on a front side and a back side of the substrate, respectively; (2) The first masking layer and the second masking layer are separately patterned to form a front nanopore window and a front reference window on the first masking layer, and a back window on the second masking layer; the back window Corresponding to the frontal nanopore window and the front reference window, the front reference window has a size larger than the front nanopore window by 200 nm to 2 μm; (3) the frontal nanopore window and the front reference The substrate corresponding to the window is subjected to a first etching to form a front reference cavity and a front nanopore cavity; (4) a tracer gas is disposed in the front nanopore cavity, and is closed; (5) corresponding to the back window Performing a second etching on the substrate while detecting whether the tracer gas is present in the working environment, and stopping the etching when detecting the presence of the tracer gas in the working environment to form the nanometer hole.

The nanopore prepared by the above preparation method can be prepared into a desired shape according to actual needs, and for example, can include a square (aspect ratio = 1:1), a rectangle (aspect ratio ≤ 10:1), and a slit (length and width) Ratio >10:1), and polygons (number of sides n>4) and so on. On this basis, different shapes of nanopores can be arrayed according to different number of mask front nanopore window arrangements.

The front side nanopore window is indicated by the front reference window, and the height of the front side reference cavity and the front side nanopore cavity formed can be ensured after etching, and the formation of such a structure can ensure that the front nanopore cavity tip is not thinned. It is etched to ensure that it is less than 1.5μm from the bottom surface after thinning, which reduces the etching time of the atomic layer etching stage and improves the preparation efficiency. Then, using atomic layer etching, etching a layer of silicon atoms in each cycle, precision control can be achieved, and high-precision tracer gas atom detection equipment can be used to perform high-precision monitoring of nanopore opening events, and nanometers can be realized. The pore size control of the pores is increased to the sub-nanometer precision level.

According to an embodiment of the present invention, the method for preparing the nanopore may further include the following additional technical features:

According to an embodiment of the invention, the front reference window in step (2) is spaced apart from the front nanopore window. The front reference window and the front nanopore window are spaced apart to facilitate the use of the front reference cavity formed by the front reference window to monitor the height of the front nanopore cavity formed by the front nanopore window, thereby facilitating the indication of the formation of the nanopore.

According to an embodiment of the present invention, between step (3) and step (4), further comprising (6): thinning the substrate corresponding to the back window to form a thinned surface, to the front reference The aperture is closest to the thinned surface to form a front reference aperture opening through the substrate.

In accordance with an embodiment of the present invention, the color feedback method is used in step (6) to monitor that the thinned surface is closest to the front reference aperture cavity to form a front reference aperture opening through the substrate. Monitoring the thinned surface by the color feedback method to communicate with the front reference cavity: adding a colored solution to the front reference cavity, when the thinned surface is thinned until the etching solution diffuses into contact with the solution At the time, the etching is stopped. By using the color feedback method to monitor, the etching can be sensitively monitored, resulting in higher precision.

According to an embodiment of the present invention, in the step (6), when the thinned surface is thinned to communicate with the front reference hole cavity, the distance between the thinned surface and the front surface nanopore cavity is less than 1.5 μm, preferably 140 nm. 1.4 μm. Thereby, the etching time of the atomic layer etching stage can be reduced, and the preparation efficiency is improved.

According to an embodiment of the invention, in the step (5), the second etching is an atomic layer etching, and the trace gas in the working environment is once detected every time one layer of atoms is etched. Atomic layer etching ensures that each cycle of etching removes an atomic layer of etched material. This further ensures the control of the preparation accuracy of the nanopore. Other etchings have no way to achieve this effect. At the same time, since the etching process is a cyclic process, the environment between each cycle is more favorable for the capture and detection of the tracer gas.

According to an embodiment of the invention, the substrate is a double throw silicon wafer, preferably a (100) crystal orientation silicon wafer, and the front side nanopore chamber and the front side reference aperture cavity are inverted quadrangular pyramid cavities.

According to an embodiment of the invention, the first masking layer and the second masking layer respectively comprise a silicon dioxide layer and a silicon nitride layer, the silicon dioxide layer is on the substrate, the silicon nitride A layer is on the silicon dioxide layer.

According to an embodiment of the invention, the first etching and the thinning treatment are performed by anisotropic wet etching.

According to an embodiment of the present invention, an anisotropic wet etching is performed using an alkaline solution which is an alkali metal hydroxide or quaternary ammonium salt. Anisotropic etching means that the etching rate of the etchant in a certain direction is much larger than other directions, and the silicon wafer is etched by using an alkali metal hydroxide such as sodium hydroxide or potassium hydroxide or a quaternary ammonium salt. You can get the structure of different graphics.

According to an embodiment of the invention, the tracer gas is enclosed in the frontal nanopore chamber using a PDMS/glass cavity structure in step (4) and then fixed.

According to an embodiment of the invention, the fixation is carried out using a device that can withstand a gas pressure of at least 0.1 MPa, the device being selected from one of an oxygen plasma surface treatment device, a bonding device or a clamp aid. The device used for fixing can guarantee a gas pressure of at least 0.1 MPa, which can ensure that the tracer gas atoms do not leak, so that precise monitoring can be achieved. When clamping by the clamping aid, the clamping force on the chip is at least greater than 2 Kg, and the application force needs to be uniform to avoid damage to the chip due to uneven force. When fixing by other means such as an oxygen plasma surface treatment apparatus or other bonding means, it is sufficient that the tracer gas atoms are not leaked, so that precise monitoring can be performed.

According to a second aspect of the invention, the invention provides a nanopore array. According to an embodiment of the invention, the nanopore arrays are arranged according to a certain rule; the nanopores are prepared by the preparation method described above. According to different needs, the nanopores can be arranged in different ways to form different nanopore arrays, which can be applied in the fields of biomacromolecule detection, near-field optics and/or atomic lithography gratings.

According to a third aspect of the invention, there is provided a device comprising a nanopore comprising the following structure:

Substrate layer,

The front side and the back side of the substrate layer are a first masking layer and a second masking layer, respectively.

The first masking layer is provided with a front reference window and a front nanohole window, and the size of the front reference window is larger than the size of the front nanohole window by 200 nm to 2 μm;

The front reference window and the front nanopore window extend toward the substrate layer to form a front reference cavity and a front nanopore cavity;

The second masking layer is provided with a back window, and the back window is disposed corresponding to the front reference window and the front nanohole window, and the back window extends toward the substrate layer to form a back cavity;

The back cavity is corresponding to the front reference cavity, and forms a front reference cavity opening;

The back cavity corresponds to the front nanopore window as a nanopore.

According to an embodiment of the present invention, the nanopore of the present invention may further include the following technical features:

According to an embodiment of the invention, the first masking layer and the second masking layer each comprise a silicon dioxide layer and a silicon nitride layer. The silicon dioxide layer is on the substrate layer, and the silicon nitride layer is on the silicon dioxide layer.

According to an embodiment of the invention, the nanopore is prepared by the preparation method described above.

According to a fourth aspect of the invention, the invention provides the use of the nanopore array described above and the device described above in the field of biomacromolecule detection, near-field optics and/or atomic lithography gratings.

The invention has the beneficial effects that the invention utilizes a high-precision tracer gas atom detecting device to perform high-precision monitoring on the nanopore opening event, and can realize the aperture control of the nanopore to be improved to the sub-nanometer precision level, thereby Sub-nano-precision silicon-based nanopores are used in the field of biomacromolecular detection to improve the precision of detection. On this basis, the substrate is precisely etched by atomic layer etching (ALE), and a layer of silicon atoms is etched in each cycle, so that precision precision can be controlled.

The additional aspects and advantages of the invention will be set forth in part in the description which follows.

DRAWINGS

The above and/or additional aspects and advantages of the present invention will become apparent and readily understood from

1 is a flow diagram of the preparation of a nanopore in accordance with one embodiment of the present invention.

2 is a schematic illustration of the height of the front side reference inverted quadrangular pyramid cavity and the front side nanopore inverted quadrangular pyramid cavity and the dimensions of the front side reference window and the front side nanopore window and the angle formed by the crystal face formed in accordance with one embodiment of the present invention.

3 is a cross-section of a clamping device that encapsulates a tracer gas structure in accordance with one embodiment of the present invention.

In the figure, the reference numerals are:

1, double throwing crystal silicon; 2, silicon dioxide layer; 3, silicon nitride layer; 4, front reference window; 5, front nanopore window; 6, front reference inverted quadrangular pyramid cavity; 7, frontal nanopore Inverted quadrangular pyramid cavity; 8, back window; 9, PDMS / glass cavity; 10, tracer gas; 11, silicon-based nanopore; 12, Teflon clamping device; 13, screw; 14, nut; Back cavity.

Detailed ways

Embodiments of the invention are described in detail below, examples of which are illustrated in the accompanying drawings. The embodiments described below with reference to the drawings are intended to be illustrative of the invention and are not to be construed as limiting.

The invention provides a preparation method of nano pores, comprising the following steps:

(1) forming a first masking layer and a second masking layer on the front and back sides of the substrate, respectively; (2) patterning the first masking layer and the second masking layer, respectively, to thereby mask the first masking layer Forming a front nanopore window and a front reference window on the layer, forming a back window on the second masking layer; the back window and the front nanopore window and the positive Correspondingly, the surface of the front reference window is 200 nm to 2 μm larger than the front nanopore window; (3) performing first etching on the substrate corresponding to the front nanopore window and the front reference window Forming a front reference hole cavity and a front surface nanopore cavity; (4) providing a tracer gas in the front surface nanopore cavity to be closed; and (5) performing a second etching on the substrate corresponding to the back surface window And simultaneously detecting whether the tracer gas exists in the working environment, and stopping the etching when detecting the presence of the tracer gas in the working environment to form the nanopore.

According to an embodiment of the present invention, the tracer gas selected in the present invention may be helium, nitrogen, argon or the like, which is chemically stable and has atomic radii of 130, 160 and 192 pm.

The size of the front reference window and the size of the front nanopore mentioned in the present invention may be any corresponding shape, and may be square, circular, or rectangular. The size of the present invention is the diameter or side length of the corresponding figure. As long as the size of the front reference window is larger than the size of the front nanopore window by 200 nm to 2 μm.

The present invention has been completed based on the following findings and findings of the inventors:

The wet etching forms nanopores. Due to the collapse structure and rectification characteristics, it is expected to fundamentally solve the problem of robustness and poor spatial and temporal resolution of nanopores. However, the existing three-step all-wet etching combined with the dry-wet method for preparing nanopores is very difficult to realize for nanopores, especially single nanopores, and it is difficult to achieve precise control of sub-nanometer resolution of nanopores. Based on the above technical problems, the inventors conducted in-depth research, and found that the precision of the nanopore preparation process can be achieved by setting a front reference window on the surface of the silicon substrate. Specifically, first, wet etching is used. The front reference cavity and the front nanopore cavity are realized on the front surface of the double-throw (100) silicon wafer, and the single-sided rapid thinning is performed by wet etching on the back surface of the silicon wafer; when the back surface is thinned to form the front reference cavity The polydimethylsiloxane (PDMS)/glass cavity structure is aligned with the front side of the silicon wafer in a tracer gas atmosphere, and reversible assembly is performed by oxygen plasma surface treatment, bonding or fixture assisting. The gas is encapsulated into the cavity; the back side of the silicon wafer is precisely etched by atomic layer etching (ALE), and after the end of each etching cycle, the presence of the tracer gas atom is performed in a vacuum environment using an internal detection instrument of the etching apparatus. Detection, when the presence of tracer gas atoms is detected, indicates the formation of the desired nanopore.

According to a specific embodiment of the present invention, the present invention provides a method for preparing a silicon-based nanopore, comprising the following steps:

(1) using a double throwing silicon wafer as a substrate, respectively forming a silicon dioxide layer on the front and back sides of the silicon wafer, and depositing a silicon nitride layer on the silicon dioxide layer;

(2) etching the silicon nitride layer and the silicon dioxide layer on the front and back sides, respectively forming a front reference window and a front nanopore window on the front side, and forming a back window on the back side, the size of the front reference window being larger than The front nanopore window is 200 nm to 2 μm in size;

(3) etching and forming a front reference inverted quadrangular pyramid cavity and a front surface nanopore inverted quadrangular pyramid cavity in the front reference window and the front nanohole window by an anisotropic wet etching method;

(4) using an anisotropic wet etching method to thin the back surface window to form a thinned surface, when thinned to Stopping etching when the reference inverted quadrangular pyramid cavity is in communication;

(5) using a PDMS/glass cavity in the tracer gas atmosphere to be aligned with the front surface, and fixed to the nanoporous inverted pyramid cavity filled with the tracer gas;

(6) performing atomic layer etching on the bottom surface after thinning, and stopping etching when the tracer gas atoms are detected;

(7) The PDMS/glass cavity is removed to obtain a sub-nanometer precision level silicon-based nanopore.

According to an embodiment of the present invention, the front reference window formed in the step (2) is 200 nm to 2 μm larger than the front nanopore window, and the formed front reference inverted quadrangular pyramid cavity and the front surface nanopore inverted quadrangular pyramid can be ensured after etching. The height of the cavity is different, and the formation of such a structure can ensure that the front nanopore quadrangular pyramid tip is not over-etched when thinning, and can be ensured to be less than 1.5 μm from the bottom surface after thinning, preferably 140 nm to 1.4 μm. Therefore, the etching time of the atomic layer etching stage is reduced, and the preparation efficiency is improved.

According to an embodiment of the present invention, in the above method for preparing a silicon-based nanopore, the step (6) uses an atomic layer etching to etch a layer of silicon atoms per cycle, thereby achieving precise precision control, and using high precision display Trace gas atom detection equipment, high-precision monitoring of nanopore opening events, can achieve nanopore aperture control to sub-nanometer accuracy level. In the preparation process, the nanopore can not directly observe the opening event of the nanopore, so it will affect the pore size of the nanopore. The tracer gas detection combined with the atomic layer etching can ensure the opening of the nanopore during the etching process. Accurate monitoring of events. At the same time, according to different precisions, the tracer gas atoms of different diameters are selected to meet the requirements of the pore size of different nanopores, and the nanopore aperture is accurately controlled.

The solution of the present invention will be explained below in conjunction with the embodiments. Those skilled in the art will appreciate that the following examples are merely illustrative of the invention and are not to be considered as limiting the scope of the invention. Where specific techniques or conditions are not indicated in the examples, they are carried out according to the techniques or conditions described in the literature in the art or in accordance with the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are conventional products that can be obtained commercially.

A specific embodiment of the present invention will be described with reference to FIG. 1, which includes the following steps:

(1) using a double throw crystal to the silicon wafer 1 as a substrate, thermally oxidizing a silicon dioxide layer 2 on both front and back sides, and depositing a silicon nitride layer 3 on the silicon dioxide layer 2; An insulating layer, a silicon nitride layer is used as a masking layer for wet etching; a silicon dioxide layer and a silicon nitride layer are collectively used as a masking layer;

(2) Using the double-sided lithography technique and the reactive ion etching technique, the silicon nitride layer 3 and the silicon dioxide layer 2 are patterned, and a front reference window 4 and a front nanopore window 5 are formed on the front side, respectively, and formed on the back surface. a back window 8, wherein the front reference window 4 has a size larger than the front nanopore window 5 by 200 nm to 2 μm;

(3) etching a front reference inverted quadrangular pyramid cavity and a front surface nanopore inverted quadrangular pyramid cavity in the front reference window and the front surface nanopore window by an alkaline etching solution anisotropic wet etching method, wherein The height difference between the front reference inverted quadrangular pyramid cavity and the front surface nanopore inverted quadrangular pyramid cavity is ΔH, and the size of the front reference window and the front nanopore window is ΔW, and the relationship is: ΔW= 2ΔH/tanα, where α is (100) crystal plane and (111) crystal The angle formed by the face, the angle of the angle is 54.74 ° (as shown in Figure 2);

(4) thinning the back window by an anisotropic wet etching method using an alkaline etching solution, and placing phenolphthalein in the solution of the inverted quadrangular pyramid cavity, and thinning the back surface of the silicon to the back cavity 15 When an event occurs, the thinning etching solution diffuses into the reference inverted quadrangular pyramid cavity to contact with the phenolphthalein, the solution turns red, and the etching is stopped. At this time, the front nanopore inverted quadrangular pyramid cavity 7 is less than 1.5 μm from the thinned surface, preferably It is 140 nm to 1.4 μm;

(5) in the tracer gas atmosphere, the PDMS / glass cavity structure 9 and the frontal nanopore inverted quadrangular pyramid cavity are aligned and fixed, so that the front nanopore inverted quadrangular pyramid cavity is filled with atmospheric pressure tracer gas 10;

(6) Inverting the fixed structure, placing it in an etching device for atomic layer etching, and turning on the built-in tracer gas atom detector to monitor the tracer atoms. After the tracer gas atom detector detects the presence of the tracer gas atom, Stop etching;

When the nanopore opening event occurs, the difference between the internal and external pressure of the PDMS/glass cavity 9 causes the tracer gas atoms in the cavity to overflow from the nanopore, and the tracer gas atom detector can detect the presence of the tracer gas atoms;

(7) The PDMS/glass cavity 9 is removed to obtain a silicon-based nanopore 11 having sub-nanometer precision controllability.

According to some embodiments of the present invention, in the above method for preparing a silicon-based nanopore, the double throwing silicon wafer is a (100) double polished wafer, and the double throwing silicon wafer has a thickness of 300 to 700 μm.

According to some embodiments of the present invention, in the above method for preparing a silicon-based nanopore, the thickness of the silicon nitride layer is

The thickness of the silicon dioxide layer is

The silica can be grown by thermal oxidation, and the silicon nitride layer is deposited by low pressure ski vapor deposition (LPCVE) to form a silicon nitride layer having a thickness of

The thickness of the silicon dioxide layer is

The silicon oxide layer can be grown by thermal oxidation using a plasma enhanced chemical vapor deposition (PECVD) silicon nitride layer to form a silicon nitride layer having a thickness of

According to some embodiments of the present invention, the etching solution is a KOH solution, and the formulation is KOH:H2O: IPA=50g: 100mL: 10mL, the etching temperature is 80° C., the precision is controlled at ±1° C., and the IPA is isopropyl. alcohol.

According to some embodiments of the present invention, the ratio (mass ratio) of the PDMS is a matrix: crosslinker = 10:1, and is cured in an environment of 80 to 120 ° C for 2 hours.

According to an embodiment of the present invention, in step (5), reversible assembly may be performed using an apparatus such as oxygen plasma surface treatment, bonding or jig assist to fix the structure. Referring to Figure 3, in accordance with an embodiment of the present invention, a Teflon holder 12 is used for attachment, wherein Figure 3 is a cross-sectional view of a clamping device enclosing a tracer gas structure, 13 being a screw and 14 being a nut . The screw 13 is fixed to the nut 14 when it is fixed, and the screw 13 is removed from the nut 14 when it is removed.

The schematic diagram of the device containing the nanopore prepared by the above preparation method is shown in Fig. 1 (7). The device can be used as a detection of biological macromolecules.

In the description of the present specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" and the like means a specific feature described in connection with the embodiment or example. A structure, material or feature is included in at least one embodiment or example of the invention. In the present specification, the schematic representation of the above terms is not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in a suitable manner in any one or more embodiments or examples. In addition, various embodiments or examples described in the specification, as well as features of various embodiments or examples, may be combined and combined.

Although the embodiments of the present invention have been shown and described, it is understood that the above-described embodiments are illustrative and are not to be construed as limiting the scope of the invention. The embodiments are subject to variations, modifications, substitutions and variations.

Claims

A method for preparing a nanopore, comprising the steps of:

(1) forming a first masking layer and a second masking layer on the front side and the back side of the substrate, respectively;

(2) patterning the first masking layer and the second masking layer respectively, thereby forming a front nanopore window and a front reference window on the first masking layer, and forming a back window on the second masking layer; The back window is disposed corresponding to the front nanopore window and the front reference window, and the size of the front reference window is 200 nm to 2 μm larger than the front nanohole window;

(3) performing a first etching on the substrate corresponding to the front nanopore window and the front reference window to form a front reference cavity and a front nanopore cavity;

(4) arranging a tracer gas in the front surface of the nanopore chamber to be closed;

(5) performing a second etching on the substrate corresponding to the back window, and detecting whether the tracer gas exists in the working environment, and stopping when detecting the presence of the tracer gas in the working environment Etching to form the nanopore.
The preparation method according to claim 1, wherein the front reference window in the step (2) is spaced apart from the front nanopore window.
The preparation method according to claim 1 or 2, further comprising (6): thinning the substrate corresponding to the back surface window between the step (3) and the step (4) A thinned face is formed until the front side reference cavity is closest to the thinned face to form a front side reference cavity opening through the substrate.
The preparation method according to claim 3, wherein in step (6), the thinned surface is monitored by a color feedback method to form a front reference hole cavity opening through the substrate closest to the front reference cavity.
The preparation method according to claim 3 or 4, wherein in the step (6), when the thinned surface is thinned to communicate with the front reference hole cavity, the thinned surface and the front surface nanopore cavity The distance is less than 1.5 μm.
The preparation method according to claim 5, wherein in the step (6), when the thinned surface is thinned to communicate with the front reference hole cavity, the distance between the thinned surface and the front surface nanopore cavity is 140 nm to 1.4 μm.
The preparation method according to any one of claims 1 to 6, wherein in the step (5), the second etching is an atomic layer etching, and each layer of atoms is etched to the working environment. The tracer gas in the test is performed once.
The preparation method according to any one of claims 1 to 7, wherein the substrate is a double throwing silicon wafer, and the front surface nanopore chamber and the front side reference hole cavity are inverted quadrangular pyramid cavities.
The method according to claim 8, wherein the substrate is a (100) crystal orientation silicon wafer.
The preparation method according to any one of claims 1 to 9, wherein the first masking layer and the second masking layer respectively comprise a silicon dioxide layer and a silicon nitride layer, the silicon dioxide a layer on the substrate, the silicon nitride A layer is on the silicon dioxide layer.
The preparation method according to any one of claims 1 to 10, wherein the first etching and the thinning treatment are performed by anisotropic wet etching.
The method according to claim 11, wherein the anisotropic wet etching is performed using an alkaline solution which is an alkali metal hydroxide or a quaternary ammonium salt.
The preparation method according to any one of claims 1 to 12, wherein in step (4), the tracer gas is enclosed in the front surface nanopore chamber by using a PDMS/glass cavity structure, and then fixed.
The preparation method according to claim 13, wherein the fixing is carried out by means of a device capable of withstanding a gas pressure of at least 0.1 MPa, the device being selected from one of an oxygen plasma surface treatment device, a bonding device or a jig auxiliary device. Kind.
A nanopore array comprising a nanopore, the nanopore being arranged according to a certain regularity;

The nanopore is prepared by the preparation method according to any one of claims 1-14.
A device containing nanopores, comprising the following structure:

Substrate layer,

The front side and the back side of the substrate layer are a first masking layer and a second masking layer, respectively.

The first masking layer is provided with a front reference window and a front nanohole window, and the size of the front reference window is larger than the size of the front nanohole window by 200 nm to 2 μm;

The front reference window and the front nanopore window extend toward the substrate layer to form a front reference cavity and a front nanopore cavity;

The second masking layer is provided with a back window, and the back window is disposed corresponding to the front reference window and the front nanohole window, and the back window extends toward the substrate layer to form a back cavity;

The back cavity is corresponding to the front reference cavity, and forms a front reference cavity opening;

The back cavity corresponds to the front nanopore window as a nanopore.
The device according to claim 16, wherein said first masking layer and said second masking layer each comprise a silicon dioxide layer and a silicon nitride layer, said silicon dioxide layer being on said substrate layer The silicon nitride layer is on the silicon dioxide layer.
The apparatus according to claim 16 or 17, wherein the nanopore is prepared by the preparation method according to any one of claims 1-14.
The use of the nanopore array of claim 15 and the device of any of claims 16-18 in the field of biomacromolecule detection, near-field optics and/or atomic lithography gratings.