Method for forming micro-pattern array by using hydrogel seal and application thereof

Abstract

A method of forming a micropattern array on a receiving substrate using a hydrogel stamp, comprising: providing a hydrogel stamp having a micropattern array disposed at the bottom thereof; loading a molecular solution into the hydrogel seal; and imprinting the molecules on a receiving substrate under the action of an electric field or current to obtain the micro-pattern array. The hydrogel seal is used under the action of an electric field or current, so that the hydrogel seal can be repeatedly imprinted, the imprinting efficiency is high, repeated infiltration is not needed, the time is saved, the cost is reduced, additional chemical reagents and complex processes are not needed, the operation is simple, the biocompatibility of a system is excellent, and the application prospect is wide. The molecular loss can be effectively reduced, and meanwhile, cross contamination in the molecular transfer printing process is avoided.

Description

A method for forming a micropattern array using a hydrogel stamp and its application Technical Field

[0001] The present application belongs to the technical field of hydrogel stamps, and in particular relates to a method for forming a micro-pattern array on a receiving substrate using a hydrogel stamp and its application. Background Art

[0002] Stamps allow molecules such as proteins to form micropattern arrays on a receiving surface, avoiding cross-contamination between patterned areas and reducing molecule waste. Hydrogel stamps are often used to simply, quickly, and economically transfer molecules, including DNA, RNA, proteins, dyes, and surfactants, onto solid surfaces. The resulting micropattern arrays can be used in high-throughput multiplexed biosensors, medical diagnostics, and drug screening. However, due to the very low diffusion rate of molecules within the hydrogel, the concentration of molecules near the stamp contact surface decreases rapidly with the number of stamps, reducing the number of uses of the hydrogel stamp and limiting its reusability.

[0003] Traditional stamps are mostly made of polydimethylsiloxane (PDMS) as the base material (Khadpekar, AJ, Khan, M., Sose, A. et al. Low Cost and Lithography-free Stamp fabrication for Microcontact Printing. Sci Rep, 2019, 9, 1024). The creation of the stamp master or template is completed using traditional micro-nano processing technology. After fabrication, the master is placed in a container with walls, and then uncrosslinked PDMS is poured onto the master. After pouring, the PDMS cures at high temperature to form a solid polymer with elasticity. The stamp is then peeled off and cut into appropriate sizes. The stamp replicates the master mold. Ink is applied to the stamp by immersion. When the stamp is in physical contact with the receiving surface substrate, the substrate will leave a patterned molecular array. However, this method requires multiple infiltrations and has low imprinting efficiency.

[0004] There are also reports of gel stamps made from agarose or acrylamide (e.g., J.Am. Chem. Soc. 2008, 130, 47, 16060-16064; Langmuir 1998, 14, 15, 3971-3975). These increase the number of impressions possible after a single infiltration of the stamp and reduce the time consumed by repeated infiltration of molecules. However, these still require multiple infiltrations to complete the impression, preventing continuous imprinting and limiting their usability.

[0005] Therefore, developing a method for patterning using a hydrogel stamp that has high imprinting efficiency, can perform continuous imprinting, has a large number of imprinting times, and does not require multiple infiltrations is a technical problem that needs to be urgently solved in this field.

[0006] Summary of the Invention

[0007] The present application provides a method for forming a micropattern array using a hydrogel stamp and its application. The hydrogel stamp, when used under the action of an electric field or current, enables repeated imprinting, resulting in high imprinting efficiency and the elimination of the need for multiple infiltrations. This saves time and reduces costs, and eliminates the need for additional chemical reagents and complex processes. The method is simple to operate and has excellent biocompatibility, effectively reducing molecular loss and avoiding cross-contamination during the molecular transfer process.

[0008] In a first aspect, the present application provides a method for forming a micropattern array on a receiving substrate using a hydrogel stamp, the method comprising:

[0009] providing a hydrogel stamp, wherein a micro-pattern array is provided on the bottom of the hydrogel stamp;

[0010] loading the hydrogel stamp with a molecular solution; and

[0011] Under the action of electric field or current, molecules are imprinted on the receiving substrate to obtain a micropattern array.

[0012] In the present application, the diffusion rate of molecules in the gel is increased under the action of an electric field or current, so that the volume of the dispersed phase is greatly increased, thereby greatly improving the number of times the hydrogel stamp can be reused. The effect of repeated imprinting can be achieved without multiple infiltrations. The imprinting efficiency is high, and continuous, automatic, rapid, and multiple imprinting are possible.

[0013] In some preferred embodiments, a molecular solution storage device is provided on the top of the hydrogel stamp.

[0014] In some preferred embodiments, an electrode is provided on the top of the hydrogel stamp.

[0015] In some preferred embodiments, the hydrogel stamp comprises a polymer material, wherein the polymer material comprises at least one of agarose, polyacrylamide, or sodium lauryl sulfate polyacrylamide.

[0016] In this application, the materials are not limited to the above-mentioned materials, and are also applicable to other materials.

[0017] In some preferred embodiments, the method for preparing the hydrogel stamp comprises:

[0018] The polymer material is mixed with a buffer solution to obtain a mixed solution; the mixed solution is poured into a mold, dried, and allowed to stand to obtain the hydrogel stamp.

[0019] In some preferred embodiments, the concentration of the hydrogel is 0.1-30%, for example, 0.2%, 0.4%, 0.6%, 0.8%, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%, 2.2%, 2.4%, 2.6%, 2.8%, 3%, 3.2%, 3.4%, 3.6%, 3.8%, 4%, 4.2%, 4.6%, 4.8%, 5%, 5.2%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.8%, 7.9%, 7.1%, 7.2%, 7.3%, 7.4%, 7.6%, 7.8%, 7. 8%, 6%, 6.2%, 6.5%, 6.8%, 7%, 7.2%, 7.5%, 7.8%, 8%, 8.2%, 8.5%, 8.8%, 9%, 9.2%, 9.5%, 9.8%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, etc.

[0020] In the present application, the concentration of the hydrogel refers to the mass volume percentage of the polymer material and the buffer solution; the concentration depends on the molecular size of the molecular material; when the molecular material is certain, too high a concentration of the hydrogel may cause the migration rate of the molecules in the hydrogel to be too slow, and too low a concentration of the hydrogel may cause the elastic modulus of the hydrogel stamp to be too low, making the stamp easy to break and the pattern easy to deform during the imprinting process.

[0021] In the present application, when a combination of polyacrylamide gel and nucleic acid is selected, the gel concentration is preferably 2-20%; when a combination of polyacrylamide gel and protein is selected, the gel concentration is preferably 2-30%; when agarose gel and DNA or protein is selected, the gel concentration is preferably 0.5-2%; when agarose gel and protein is selected, the gel concentration is preferably 0.5-10%.

[0022] For example, when a combination of agarose gel and DNA is selected, and the gel concentration is less than 0.5%, the resulting hydrogel stamp has a low elastic modulus, the stamp is easily broken, and even causes pattern deformation; when the concentration is greater than 2%, the migration rate of DNA molecules is slow and the imprint is unclear.

[0023] In some preferred embodiments, the mixing temperature is 20-100°C, for example, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, 95°C, etc.

[0024] In this application, when the gel material is agarose, the temperature is preferably 90-100°C; when the gel material is polyacrylamide, the temperature is preferably 20-30°C.

[0025] In some preferred embodiments, the pH value of the mixture is 6-8.

[0026] In some preferred embodiments, the height of the mold is 1 μm to 10 mm, for example, 2 μm, 5 μm, 10 μm, 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, etc.

[0027] In this application, if the mold height is too high, it may take too long for the DNA molecules to fill the hydrogel; if the height is too low, the hydrogel may easily fall out of the mold. In some preferred embodiments, the mold opening is covered with a patterned PDMS master. The master can be manufactured using any suitable method, such as standard micro-nanofabrication technology.

[0028] In some preferred embodiments, the drying is performed under vacuum conditions.

[0029] In some preferred embodiments, the standing temperature is 3-5°C, for example, 3°C, 4°C, 5°C, etc.

[0030] In some preferred embodiments, the standing time is 0.5 to 3 hours, for example, 1 hour, 1.5 hours, 2 hours, 2.5 hours, etc.

[0031] In some preferred embodiments, the molecular solution includes at least one of a DNA molecule solution, an RNA molecule solution, a protein solution, a dye, or a surfactant solution.

[0032] In some preferred embodiments, the concentration of the molecular solution is 0.1 to 3 mol / L, for example, 0.2 mol / L, 0.5 mol / L, 0.8 mol / L, 1 mol / L, 1.5 mol / L, 2 mol / L, 2.5 mol / L, etc.

[0033] In some preferred embodiments, the solvent of the molecular solution comprises a buffer.

[0034] In some preferred embodiments, the method of loading the molecular solution into the hydrogel stamp comprises: injecting the molecular solution into the molecular solution storage device, and applying an electric field until the molecular solution is completely diffused into the hydrogel stamp.

[0035] In some preferred embodiments, the time for applying the electric field is 0.5 to 5 h, for example, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, etc.

[0036] In some preferred embodiments, the method of imprinting under the action of an electric field comprises: disposing an electrode on the top of the hydrogel stamp, disposing an electrode plate under the imprinted receiving substrate, and imprinting in the electric field to obtain a micropattern array.

[0037] In some preferred embodiments, the electric field strength is 4 to 15 V / cm, for example, 5 V / cm, 6 V / cm, 7 V / cm, 8 V / cm, 9 V / cm, 10 V / cm, 11 V / cm, 12 V / cm, 13 V / cm, 14 V / cm, 15 V / cm, etc.

[0038] In the present application, when the electric field strength is within a specific range, the higher the efficiency of the hydrogel stamp imprinting, the higher the accuracy, and the clearer and more complete the pattern; when the electric field strength is less than 4V / cm, the molecular migration rate is slow and the imprinting efficiency is low; when the electric field strength is greater than 15V / cm, the imprinted pattern is prone to deformation or dislocation and has low accuracy.

[0039] In some preferred embodiments, the surface of the receiving substrate is further provided with a silane coating.

[0040] In some preferred embodiments, the silane includes at least one of 3-aminopropyltriethoxysilane, 3-aminopropyltrimethoxysilane, or 3-methacryloxypropyltrimethoxysilane.

[0041] In some preferred embodiments, the method of imprinting under the action of electric current includes: providing an electrode on the top of the hydrogel stamp, connecting the receiving substrate and the electrode on the top of the hydrogel stamp to a power source respectively, and imprinting under the action of electric current to obtain a micropattern array; the receiving substrate is a conductive substrate.

[0042] Preferably, the current intensity is 0.4-2 mA / cm, for example, 0.5 mA / cm, 0.6 mA / cm, 0.8 mA / cm, 1 mA / cm, 1.2 mA / cm, 1.4 mA / cm, 1.6 mA / cm, 1.8 mA / cm, etc.

[0043] Within the specific range of current intensity, the hydrogel stamp has higher efficiency and higher precision, and the pattern is cleaner and more complete. When the current intensity is less than 0.4 mA / cm, the molecular migration rate is slow and the stamping efficiency is low. When the current intensity is greater than 2 mA / cm, the thermal effect causes DNA and protein denaturation. In addition, the evaporation of water caused by the thermal effect can also cause the hydrogel stamp to deform, and the stamped pattern is also prone to deformation or dislocation, with low precision.

[0044] In the present application, the conductive substrate may be a substrate material having a conductive function itself, or a substrate material having a conductive coating on its surface.

[0045] In some preferred embodiments, the conductive substrate includes a glass substrate and a conductive layer stacked sequentially.

[0046] In the present application, the conductive layer is close to the side of the hydrogel stamp, and there is no excessive restriction on the material of the conductive layer. A variety of materials can be used, such as metal materials such as gold, platinum, and silver, non-metallic materials such as graphite and conductive polymers, or conductive mixtures of metals and non-metals.

[0047] In some preferred embodiments, a silane coating is further provided between the glass substrate and the conductive layer.

[0048] In some preferred embodiments, the silane includes at least one of 3-aminopropyltriethoxysilane, 3-aminopropyltrimethoxysilane, or 3-methacryloxypropyltrimethoxysilane.

[0049] In some preferred embodiments, the interval time of the stamping is 1 to 30 s, for example, it can be 2 s, 4 s, 6 s, 8 s, 10 s, 12 s, 14 s, 16 s, 18 s, 20 s, 22 s, 24 s, 26 s, 28 s, etc.

[0050] As a preferred technical solution of the present application, the method comprises: placing a molecular solution storage device and an electrode on the top of a hydrogel stamp with a hydrogel concentration of 0.1 to 30%, placing an electrode plate under the imprinted receiving substrate, and imprinting under an electric field strength of 4 to 15 V / cm to obtain a micropattern array; a silane coating is provided between the receiving substrate and the hydrogel stamp; and the time interval of the imprinting is 1 to 30 seconds;

[0051] Alternatively, a molecular solution storage device and electrodes are set on the top of a hydrogel stamp with a hydrogel concentration of 0.1 to 30%. The receiving substrate and the electrodes on the top of the hydrogel stamp are respectively connected to a power supply, and imprinting is performed under the action of current to obtain a micropattern array; the receiving substrate is a conductive substrate; and the time interval of the imprinting is 1 to 30 seconds.

[0052] In the present application, the positive and negative electrodes on the top of the hydrogel stamp and the electrodes below the receiving substrate are determined by the positive and negative electrodes of the molecular solution; for example, when the molecular charge is positive, a positive electrode is set on the top of the hydrogel stamp or connected to the positive electrode of the power supply, and the direction of the electric field should be from the top of the hydrogel stamp toward the bottom of the hydrogel stamp (i.e., the direction of the receiving substrate); when the molecular charge is negative, a negative electrode is set on the top of the hydrogel stamp or connected to the negative electrode of the power supply (the conductive layer has opposite charge to the electrode on the top of the hydrogel stamp), and the direction of the electric field should be from bottom to top, i.e., from the receiving substrate to the hydrogel stamp.

[0053] In the present application, examples of negatively charged molecules include DNA or RNA molecules, especially DNA or RNA molecules in a buffer solution at pH 7; examples of positively charged molecules include trypsin, especially trypsin in a buffer solution at pH 7.

[0054] In a second aspect, the present application provides an application of the method according to the first aspect in the fields of biosensors, medical diagnosis or drug screening.

[0055] The numerical range described in this application includes not only the point values ​​listed above, but also any point values ​​between the above numerical ranges that are not listed. Due to limited space and for the sake of simplicity, this application no longer exhaustively lists the specific point values ​​included in the range.

[0056] Compared with the prior art, the present invention has the following advantages:

[0057] The method for using a hydrogel stamp provided in the present application can effectively reduce molecular loss and avoid cross-contamination during the molecular transfer process by using the hydrogel stamp for imprinting under the action of an electric field or current. The method also increases the diffusion rate of molecules within the hydrogel and can control the direction of the electric field between plates according to the different electrical properties of the molecules, greatly improving the single-processing utilization rate of the hydrogel stamp, improving transfer production efficiency and reducing costs. In addition, the present application is a physical process that does not require additional chemical reagents or complex processes, is simple to operate, and has excellent biocompatibility of the system. It has few material restrictions, is easy to mass-produce, and can be achieved through automation, greatly improving imprinting efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] FIG1 is a schematic diagram of the method provided in Example 2;

[0059] Among them, 1-positively charged electrode plate; 2-hydrogel stamp with positively charged molecules; 3-glass substrate coated with 3-(methacryloyloxy)propyltrimethoxysilane; 4-negatively charged electrode plate; after imprinting, molecular imprints are left on the substrate surface;

[0060] FIG2 is a schematic diagram of the method provided in Example 4;

[0061] Among them, 1-electrode plate connected to the positive pole of the power supply; 2-hydrogel stamp loaded with positively charged molecules; 3-glass substrate coated with 3-(methacryloyloxy)propyltrimethoxysilane; 4-conductive layer; 5-power supply; 6-wire; after imprinting, a molecular imprint is left on the surface of the substrate. DETAILED DESCRIPTION

[0062] The technical solution of the present application is further described below through specific implementation methods. Those skilled in the art should understand that the embodiments are only used to help understand the present application and should not be regarded as specific limitations of the present application.

[0063] Example 1: Imprinting using a hydrogel stamp

[0064] (1) Preparation of hydrogel stamps

[0065] Agarose powder and TAE buffer (40mM Tris-acetate (SIGMA), 1mM EDTA (SIGMA)) were weighed into a conical flask and thoroughly mixed at 95°C to obtain an agarose solution. The pH of the mixed solution was maintained between 6 and 8, and the gel concentration was 1.0%. The thoroughly mixed agarose solution was immediately poured into a stamp mold (1mm high, 3mm wide, and 6mm long). After the agarose solution was poured into and filled the mold, the mold opening was covered with a patterned PDMS master. The mold containing the agarose solution was then placed in a desiccator connected to a vacuum pump to remove air bubbles. The mixture was allowed to stand at 4°C for 3 hours to obtain a patterned hydrogel stamp.

[0066] (2) Ink injection

[0067] DNA molecules with thiol groups at the ends and a molecular weight of 2000 to 5000 bp were mixed with TAE buffer to obtain a DNA molecule solution with a concentration of 1 mg / ml. The DNA molecule solution was injected into a molecular solution storage device on top of a hydrogel stamp. The hydrogel stamp was placed with the patterned side facing down into an electric field device with a positive electrode plate underneath. The DNA molecules were allowed to diffuse completely into the hydrogel stamp for 3 hours.

[0068] (3) Imprinting

[0069] The PDMS master on the mold was peeled off, and a glass substrate coated with 3-(methacryloyloxy)propyltrimethoxysilane was placed between the positive electrode plate and the hydrogel stamp. The patterned side of the hydrogel stamp was facing the positive electrode plate, and the hydrogel stamp was imprinted in an electric field with an electric field strength of 5 V / cm.

[0070] Example 2

[0071] This example was carried out according to the method of Example 1. The only difference from Example 1 was that the gel concentration in step (1) was 0.5%, the DNA molecule solution was replaced with an equal concentration of 24 kDa trypsin buffer solution in step (2), and the electric field strength in step (3) was 6 V / cm. Other materials, amounts, and step parameters were the same as those in Example 1.

[0072] Example 3

[0073] This example was carried out according to the method of Example 1. The only difference from Example 1 was that the gel concentration in step (1) was 1.5%, and the electric field strength in step (3) was 4 V / cm. Other materials, amounts, and step parameters were the same as those in Example 1.

[0074] Example 4

[0075] This embodiment provides a method for using a hydrogel stamp, which specifically includes the following steps:

[0076] Steps (1) and (2) are the same as those in Example 2;

[0077] (3) Imprinting

[0078] An electrode plate connected to the positive pole of a power supply is provided on the top of the hydrogel stamp. The conductive substrate includes a conductive layer (made of graphite), a 3-(methacryloyloxy)propyltrimethoxysilane coating, and a glass substrate arranged in sequence. The conductive substrate is placed under the hydrogel stamp with the patterned side of the hydrogel stamp facing the conductive substrate. The conductive layer is connected to the negative pole of the power supply, and the hydrogel stamp is imprinted under the action of electric current.

[0079] Example 5

[0080] This embodiment is carried out according to the method of Example 1. The only difference from Example 1 is that the electric field intensity in step (3) is 20 V / cm. Other materials, amounts and step parameters are the same as those in Example 1.

[0081] Example 6

[0082] This embodiment is carried out according to the method of Example 1. The only difference from Example 1 is that the electric field intensity in step (3) is 1 V / cm. Other materials, amounts and step parameters are the same as those in Example 1.

[0083] Example 7

[0084] This example was carried out according to the method of Example 1. The only difference from Example 1 was that the gel concentration in step (1) was 15%. Other materials, amounts and step parameters were the same as those in Example 1.

[0085] Example 8

[0086] This example was carried out according to the method of Example 1. The only difference from Example 1 was that the gel concentration in step (1) was 0.05%. Other materials, amounts and step parameters were the same as those in Example 1.

[0087] Therefore, the method for using the hydrogel stamp provided in the present application can improve the imprinting efficiency and achieve continuous imprinting by imprinting the hydrogel stamp under the action of an electric field or current, without the need for multiple infiltrations in molecular solutions, thus saving process flow, time and cost, improving the imprinting efficiency, and the obtained pattern is clear and complete.

[0088] When the process parameters are not specified in the present application, such as in Examples 5 to 8, the embossing effect is poor, the precision is low, and the pattern may be deformed or unclear.

[0089] In summary, the usage method provided by the present application uses the hydrogel stamp for imprinting under the action of an electric field or current, which can effectively reduce the loss of molecules and avoid cross-contamination during the molecular transfer process; it increases the diffusion rate of molecules in the hydrogel, and can control the direction of the electric field between plates according to the different electrical properties of the molecules, greatly improving the single-processing utilization rate of the hydrogel stamp, improving the transfer production efficiency and reducing costs; in addition, the present application is a physical process, does not require additional chemical reagents and complex processes, is simple to operate, and the system has excellent biocompatibility, has little material restriction, is easy to mass-produce, and can be achieved through automation.

[0090] The applicant declares that the above is only a specific implementation method of the present application, but the protection scope of the present application is not limited thereto. Technical personnel in the relevant technical field should understand that any changes or substitutions that can be easily thought of by technical personnel in the relevant technical field within the technical scope disclosed in this application fall within the protection scope and disclosure scope of this application.

Claims

1. A method for forming a micropattern array on a receiving substrate using a hydrogel stamp, comprising: providing a hydrogel stamp, wherein a micro-pattern array is provided on the bottom of the hydrogel stamp; loading a molecular solution into the hydrogel stamp; as well as Under the action of electric field or current, molecules are imprinted on the receiving substrate to obtain a micropattern array.

2. The method according to claim 1, wherein The hydrogel stamp contains a polymer material, and the polymer material includes at least one of agarose, polyacrylamide or sodium lauryl sulfate polyacrylamide.

3. The method according to claim 1 or 2, wherein A molecular solution storage device is provided on the top of the hydrogel stamp.

4. The method according to any one of claims 1 to 3, wherein An electrode is provided on the top of the hydrogel stamp.

5. The method according to any one of claims 1 to 4, wherein The preparation method of the hydrogel stamp comprises: Mixing a polymer material with a buffer solution to obtain a mixed solution; pouring the mixed solution into a mold, drying, and allowing to stand to obtain the hydrogel stamp; Wherein, the polymer material is at least one of agarose, polyacrylamide or sodium lauryl sulfate polyacrylamide; Preferably, the concentration of the hydrogel in the hydrogel stamp is 0.1-30%.

6. The method according to claim 5, wherein: The mixing temperature is 20 to 100° C. Preferably, the pH value of the mixture is 6 to 8; Preferably, the height of the mold is 1 μm to 10 mm; Preferably, the drying is carried out under vacuum conditions; Preferably, the standing temperature is 3-5°C; Preferably, the standing time is 0.5 to 3 hours.

7. The method according to any one of claims 1 to 6, wherein The molecular solution includes at least one of a DNA molecular solution, an RNA molecular solution, a protein solution, a dye or a surfactant solution.

8. The method according to any one of claims 1 to 7, wherein The concentration of the molecular solution is 0.1 to 3 mol / L; Preferably, the solvent of the molecular solution comprises a buffer.

9. The method according to any one of claims 1 to 8, wherein The method of loading the molecular solution into the hydrogel stamp comprises: injecting the molecular solution into the molecular solution storage device, applying an electric field until the molecular solution is completely diffused into the hydrogel stamp; Preferably, the electric field is applied for a time of 0.5 to 5 hours.

10. The method according to any one of claims 1 to 9, wherein The method of imprinting under the action of an electric field comprises: arranging an electrode on the top of a hydrogel stamp, arranging an electrode plate below the imprinted receiving substrate, and imprinting in an electric field to obtain a micro-pattern array; Preferably, the electric field strength is 4 to 15 V / cm.

11. The method according to any one of claims 1 to 10, wherein The receiving substrate surface is also provided with a silane coating; Preferably, the silane includes at least one of 3-aminopropyltriethoxysilane, 3-aminopropyltrimethoxysilane or 3-methacryloxypropyltrimethoxysilane.

12. The method according to any one of claims 1 to 9, wherein The method for imprinting under the action of electric current comprises: providing an electrode on the top of a hydrogel stamp, connecting the receiving substrate and the electrode on the top of the hydrogel stamp to a power source respectively, and imprinting under the action of electric current to obtain a micro-pattern array; the receiving substrate is a conductive substrate; Preferably, the current intensity is 0.4-2 mA / cm.

13. The method according to claim 12, wherein: The conductive substrate comprises a glass substrate and a conductive layer stacked in sequence; Preferably, a silane coating is further provided between the glass substrate and the conductive layer; Preferably, the silane comprises at least one of 3-aminopropyltriethoxysilane, 3-aminopropyltrimethoxysilane or 3-methacryloxypropyltrimethoxysilane; Preferably, the interval time of the stamping is 1 to 30 seconds.

14. The method according to any one of claims 1 to 13, wherein: The method comprises: placing a molecular solution storage device and an electrode on the top of a hydrogel stamp with a hydrogel concentration of 0.1 to 30%, placing an electrode plate under an imprinted receiving substrate, and performing imprinting under an electric field strength of 4 to 15 V / cm to obtain a micro-pattern array; a silane coating is provided between the receiving substrate and the hydrogel stamp; and the time interval of the imprinting is 1 to 30 seconds; Alternatively, a molecular solution storage device and an electrode are provided on the top of a hydrogel stamp with a hydrogel concentration of 0.1 to 30%. The receiving substrate and the electrode on the top of the hydrogel stamp are respectively connected to a power source, and imprinting is performed under the action of current to obtain a micropattern array; the receiving substrate is a conductive substrate; and the time interval of the imprinting is 1 to 30 seconds.

15. Use of the method according to any one of claims 1 to 14 in the fields of biosensors, medical diagnosis or drug screening.

Images