KR20100114238A - The highly sensitive pre-patterned micro array chip for bio-molecules detection with hydrophobic and hydrophilic surface and fabrication method thereof - Google Patents

Abstract

PURPOSE: An array chip of high sensitivity for bio molecule analysis using hydrophobic and hydrophilic surfaces and a method for manufacturing the same are provided to intensively arrange inside a microwell. CONSTITUTION: An array chip(10) of high sensitivity comprises a plurality of microwells(3) on a substrate(1). The microwell has metal thin film(5) and hydrophobic(7) in laminated multilayer thin film. The metal thin film has Ti, Cr, Al, Cu, Au, Ag, Pt, Si, Rh, Ru, V, Ta or mixture thereof. A method for manufacturing the microarray comprises: a step of depositing the metal thin film on the substrate; a depositing hydrophobic thin film on the metal thin film; a step of patterning the metal thin film and hydrophobic thin film by picture process; a step of etching the metal thin film and hydrophobic thin film to form a plurality of microwells; and a step of forming hydrophilic thin film with hydrophilic functional group.

Description

The highly sensitive pre-patterned micro array chip for bio-molecules detection with hydrophobic and hydrophilic surface and fabrication method

The present invention relates to a highly sensitive array chip and a method for manufacturing the same, which can be intensively aligned in a microwell into which a sample is injected, and have an excellent signal-to-noise ratio to obtain reliable analysis results.

Protein chip is a biochip technology, which refers to a biochip that finds protein information by immobilizing related substances such as antibodies, receptors, ligands, nucleic acids, and carbohydrates that can react with a specific protein at a high density on a single chip.

Protein chips are manufactured by aligning and attaching proteins with known structures and functions to small glass, plastic, or metal substrates, which require stable immobilization and alignment of proteins on the substrate.

Various methods are used for the immobilization of proteins, and the surface treatment method of the substrate to increase the affinity of the protein with the substrate is the most common.

As a protein chip substrate, a well plate using a plastic material, a glass slide, a porous gel pad slide, and the like are typically used.

In the case of glass slides, the formation of amine bonds on the substrate surface is commonly used when proteins are fixed on the substrate.

In addition, in the case of a metal substrate, a self-assembly method may be used to increase the affinity of only the immobilized portion of the surface in order to use microstamping capable of selective patterning. The intermediate mediator between the receptor and the solid substrate can be used to control the orientation of the protein during protein immobilization.

However, the surface-treated substrate reacts with oxygen and moisture in the atmosphere during storage and distribution periods before production and use, resulting in a decrease in substrate activity.

Korean Patent Laid-Open Publication No. 2006-17267 proposes a microarray that attaches a UV film to a surface of a substrate to which an active group or a probe molecule is attached on a surface to prevent a decrease in adhesion when exposed to ultraviolet rays. However, in this method, a process is added due to UV film production, substrate adhesion, and the like, and there is a concern that the phenomenon of lifting between the substrate and the UV film may occur due to external environment.

Moreover, protein chips have many problems, such as instability of the protein itself, spatial disturbance due to the three-dimensional structure, and the possibility of cross-contamination between each point.

On the other hand, the analysis using the protein chip is performed through an optical method. In this case, a plastic well plate may require a large amount of sample for analysis, a low signal-to-noise ratio, and a low analysis sensitivity.

In addition, in the case of glass slides, an expensive device is required for a separate arrangement. In addition to volatilization of a sample, cross contamination between adjacent samples occurs, which also causes a poor signal-to-noise ratio, and uniform analysis image. There is a problem that can not.

In the case of using a substrate coated with a porous gel pad, the signal-to-noise ratio is good, but residues remain after washing and a large amount of sample is required for analysis.

In order to solve the above problems, an object of the present invention is to provide a highly sensitive array chip and a method of manufacturing the same, which is easy to align the sample and has an excellent signal-to-noise ratio.

In order to achieve the above object,

Having a plurality of microwells on a substrate;

The microwell provides a highly sensitive array chip formed of a multilayer thin film in which a metal thin film and a hydrophobic thin film are stacked.

In addition, the high sensitivity array chip provides a high sensitivity array chip in which a hydrophilic thin film having a hydrophilic functional group is formed on a surface of a substrate inside an opening of a microwell.

In addition,

Depositing a metal thin film on the substrate;

Depositing a hydrophobic thin film on the metal thin film;

Patterning the metal thin film and the hydrophobic thin film through a photolithography process; And

A method of manufacturing a highly sensitive array chip comprising etching a metal thin film and a hydrophobic thin film on which the pattern is formed to form a plurality of microwells is provided.

In addition, the high sensitivity array chip further performs a step of forming a hydrophilic thin film having hydrophilic functional groups on the surface of the substrate inside the opening of the microwell after the microwell forming step.

The highly sensitive array chip according to the present invention can be intensively aligned inside a microwell into which a sample is injected, has an excellent signal-to-noise ratio, obtains reliable analysis results, and can be applied for biological analysis.

The present invention is described in more detail below.

1 and 2 are a cross-sectional view and a three-dimensional view showing a high sensitivity array chip according to a first embodiment of the present invention.

1 and 2, the highly sensitive array chip 10 includes a plurality of microwells 3 on a substrate 1.

The microwells 3 are vertically and horizontally disposed on the substrate 1 at equal intervals, and various shapes of a cylindrical shape, a polyhedron, an inverted cone shape, or an inverted pyramid shape are possible. 10 to 10000 of these microwells 3 are formed on the substrate 1, and the number and size of the microwells 3 may vary depending on the application.

The substrate 1 may be a glass substrate, a semiconductor substrate or a plastic substrate and is not particularly limited in the present invention. Typically, semiconductor substrates are Si, Ge, C, Ga, As, P, B, Zn, Se, S, Cd, Sn, Al, In, SiGe, GaAs, AlGaAs, GaAsP, InAs, Sn, InAsP, InGaAs, AlAs, InP, GaP, ZnSe, CdS, ZnCdS, CdSe and one selected from the group consisting of a combination thereof are possible. In addition, the plastic substrate includes one material selected from the group consisting of ester polymers, silicone polymers, acrylic polymers, olefin polymers, and copolymers thereof. Preferably polyethylene terephthalate (PET), polybutylene terephthalate (PBT), polysilane, polysiloxane, polysilazane, polyacrylate, polymethylmethacrylate, polyethylacrylate, polyethylmethacrylate, inter Click olefin polymers, polyethylene, polypropylene, polyimide, polystyrene, polyacetal, polyetheretherketone, polyestersulfone, polytetrafluoroethylene, polyvinylchloride, polyvinylidene fluoride, perfluoroalkyl polymers and their One selected from the group consisting of a combination is possible.

In particular, in the present invention, the microwell 3 is formed of a multilayer thin film in which the metal thin film 5 and the hydrophobic thin film 7 are stacked.

The metal thin film 5 can selectively suppress the optical signal and reduce the background signal, and is formed to a thickness of 1 nm to 1000 nm.

The metal thin film 5 may be made of any conventional metal material, and is typically Ti, Cr, Al, Cu, Au, Ag, Pt, Si, Rh, Ru, V, Ta, or a combination thereof. It includes one selected from the group consisting of, and preferably using Ti.

The hydrophobic thin film 7 allows the hydrophilic sample solution to be analyzed to be well aligned only in the analysis part, that is, inside the microwell 3, without getting wet in the background part of the microwell 3. The material of such a hydrophobic thin film 7 may be any material as long as the material exhibits hydrophobicity.

For example, the hydrophobic thin film 7 may include perfluoro trichlorosilane (FOTS), octadecyl mercaptan (ODT, Octadecylmercaptan), trichlorooctadecylsilane (OTS, Trichlorooctadecylsilane), and dichlorodimethylsilane (DDMS , Dichlorodimethylsilane (DTS), Undecyltrichlorosilane (DTS), Undecenyltrichlorosilane (Undecenyltrichlorosilane), Perfluorodecyltrichlorosilane (PDTS, Perfluorodecyltrichlorosilane), Trichlorovinylsilane (TVS, Trichlorovinylsilane), Perfluoro It includes one species selected from the group consisting of decanoic acid (PFDA, Perfluorodecanoic Acid) and combinations thereof.

The hydrophobic thin film 7 is formed to a thickness of 1nm to 10nm, preferably formed of a self-assembled thin film (SAM). Self-assembled molecular thin film has a relatively long alkyl group, and self-assembly of two-dimensional alignment on the surface of the substrate by using molecules having functional groups capable of covalent bonding by interacting with the surface of the substrate. By using this means a monolayer aligned (monolayer). In the present invention, the hydrophobic thin film 7 is formed of a self-assembled molecular thin film to have a very stable and uniform surface.

In addition to the self-assembled molecular thin film, the hydrophobic thin film 7 may contain source gases such as CHF ₃ , CH ₂ F ₂ , C ₂ F ₆ , C ₃ F ₈ , C ₄ F ₈ , C ₅ F ₈ , and C ₄ F ₆ . It can be used by depositing by plasma chemical vapor deposition method.

The hydrophobic thin film 7 is also a photoresist used in a photographic process, typically a polyvinylphenol polymer, a polyhydroxystyrene polymer, a polynorbornene polymer, a polyadamant polymer, a polyimide polymer, a polyacryl One kind selected from the group consisting of a rate-based polymer, a polymethacrylate-based polymer, a polyfluorine-based polymer, and a combination thereof is possible.

In addition, hydrophobic and chemical-resistant fluororesins such as polytetrafluoroethylene (PTFE), polychlorotrifluoroethylene (PCTFE), polyvinylidene fluoride (PVDF) and polyvinyl fluoride (PVF) Can be used.

Here, aromatic hydrocarbons, typically 2-tert-butyl-9,10-di (2-naphthyl) anthracene, anthracene, 9,10-diphenylanthracene, tetracene, rubin, perylene, parylene N ( Parylene N), Parylene C, Parylene CF, Parylene D, Parylene HT, Parylene SR, Parylene SF ), 2,5,8,11-tetra (tert-butyl) peryline, pentacene, coronine, and a combination thereof.

In addition, polydimethylsiloxane group, trade names Apical ^® ,, Kapton ^® ,, Norton TH ^® ,, Kaptrex ^®, Phosphorus linear polyimides, Aromatic heterocyclic polyimides And one selected from the group consisting of a combination thereof.

These materials may form the hydrophobic thin film 7 through a conventional wet coating method, and may be formed by various methods such as a flow coating method, a spin coating method, and a dip coating method.

3 is a cross-sectional view illustrating a high sensitivity array chip according to a second embodiment of the present invention.

Referring to FIG. 3, in the highly sensitive array chip 10, a hydrophilic thin film 9 is formed on the surface of the substrate 1 in the opening region of the microwell 3, thereby modifying the surface to be hydrophilic.

In other words, hydrophilicity on the surface of the substrate 1 on which the metal thin film 5 / hydrophobic thin film 7 is not formed, i.e., on the surface of the lower substrate 1 in the opening of the microwell 3, the analysis portion into which the sample is directly injected. The hydrophilic thin film 9 having a functional group is formed.

The hydrophilic functional group is selected from the group consisting of hydroxy group (-OH), amine group (-NH ₂ ), aldehyde group (-C (= O) H), carboxyl group (-C (= O) OH) and combinations thereof It includes one kind. In the case of proteins or nucleic acids, the main component is an amino acid, and proteins and nucleic acids introduced into a sample to be analyzed are stably fixed to the substrate 1 due to hydrophilic functional groups present on the surface of the substrate 1.

Preferably, the hydrophilic thin film 9 is R ^1- (CH ₂ ) n -Si- (R ² ) ₃ , wherein R ¹ = OH, NH ₂ , COH, or COOH, and R ² is Cl or OC _n Alkoxy of H _{2n + 2} , n is 1 to 6) trichlorosilane or trialkoxysilane is possible, more preferably, aminopropyltriethoxysilane (3-aminopropyltriethoxysilane), aminoethylaminopropyl Trimethoxysilane (N- (2-aminoethyl) -3-aminopropyltrimethoxysilane), aminotrimethoxysilane ((3-aminopropyl) trimethoxysilane), aminopropyldiethoxymethylsilane (3-Aminopropyl (diethoxy) methylsilane), trie One kind from the group consisting of (3-triethoxysilylpropyl) succinic anhydride and combinations thereof is possible.

Such a compound having a hydrophilic functional group is chemically reacted with OC _n H _{2n + 2} , which is a chloro group or an alkoxy group in a molecule, and oxygen present in the substrate, thereby being covalently bonded to the substrate to be stably fixed to the substrate. SAM).

In addition, depending on the object to be analyzed, the surface of the hydrophilic thin film 9 can be modified with other suitable hydrophilic functional groups. For example, in the embodiment of the present invention, the surface of the hydrophilic thin film having an amine group with aminopropyltriethoxysilane may be modified with glutaaldehyde to react the amine group with glutaaldehyde so that the surface has an aldehyde group.

The material for such a surface treatment is not particularly limited in the present invention, and any material may be used as long as it is a material known in the art such as glutaraldehyde and acetic acid.

The manufacture of the highly sensitive array chips as described above is carried out in various deposition and coating processes.

The highly sensitive array chip according to the first embodiment of the present invention

(S1) depositing a metal thin film on the substrate;

(S2) depositing a hydrophobic thin film on the metal thin film;

(S3) patterning the metal thin film and the hydrophobic thin film through a photolithography process; And

(S4) The metal thin film and the hydrophobic thin film on which the pattern is formed are manufactured by etching through an etching process to form a microwell.

Each step will be described in more detail below. 4 to 8 are cross-sectional views illustrating a manufacturing procedure of the high sensitivity array chip.

First, as shown in FIG. 4, the metal layer 2 is deposited on the substrate 1 (S1).

The deposition is not limited in the present invention, it is carried out using a conventional deposition method. Representatively, it is performed by one method selected from the group consisting of sputtering, electron beam deposition, thermal deposition, laser molecular beam deposition, pulse laser deposition, metal organic chemical vapor deposition, chemical vapor deposition, and plasma deposition.

Next, a hydrophobic thin film layer 4 is deposited on the metal layer 2 (FIG. 5, S2).

The hydrophobic thin film layer 4 is formed using a material as described above using a self-assembled thin film formation method, a plasma enhanced chemical vapor deposition method, and a conventional wet coating method. The specific method may be appropriately selected by those skilled in the art according to the respective materials.

For example, the hydrophobic thin film layer 4 in the form of a self-assembled thin film is prepared by treating oxygen metal to the metal layer 2 by introducing an oxygen group and introducing a material as stated in the activated hydroxy functional group into a vapor phase. It is possible.

Next, the metal layer 2 and the hydrophobic thin film layer 4 are patterned and etched using an optical lithography method to form a microwell 3 made of a multilayer thin film of the metal thin film 5 / hydrophobic thin film 7. (S3).

The optical lithography method is not particularly limited in the present invention, and through a conventional photolithography process such as photoresist coating-> soft bake-> exposure-> development-> hard bake-> etching-> strip process as is well known. To perform.

For example, as shown in FIG. 6, the photoresist 6 is applied over the entire surface of the hydrophobic thin film layer 4.

The application of the photoresist 6 may be a conventional wet coating method, typically a dip coating, a doctor blade method, a spin coating, a spray coating, a slit coating, a casting method, or a lamination method. Etc. The photoresist 6 is not particularly limited in thickness, and is generally made to be 1 μm to 100 μm or less.

Subsequently, a mask is placed on the coated photoresist 6 film and then exposed. As the mask, a Cr mask, an emulsion mask, a film mask, or the like may be used, and an appropriate one may be selected according to the minimum line width and lifetime to be processed. At this time, the exposure process uses general UV (360 ~ 450nm), KrF (248nm), ArF (193nm), VUV (157nm), EUV (13nm), E-beam, X-ray or ion beam as the exposure source, 0.1 Preferably, the exposure is performed at an exposure energy of 50 mJ / cm ² .

Referring to FIG. 7, the photoresist 6 film is treated with the post-exposure developer to remove other photoresist 6 except for the exposed portion to form a pattern.

The developing process refers to a process of melting a photoresist 8 in a portion where bonding is relatively weak through an exposure process with a developing solution, and through this process, a metal thin film 5 / hydrophobic thin film (patterned on the substrate 1). A microwell 3 made of a multilayer thin film of 7) is formed (FIG. 8).

At this time, the developer used may be carried out using an alkaline developer that is commonly used, it is preferable that the alkaline developer is an aqueous solution of tetramethylammonium hydroxide (TMAH) of 0.01 to 5% by weight.

The development process refers to a process of melting a hydrophobic thin film and a metal thin film with an etchant, and through this process, a microwell (3) consisting of a multilayer thin film of a metal thin film (5) / hydrophobic thin film (7) patterned on a substrate (1). To form (Fig. 8, S4).

At this time, the etching solution to be used is preferably selected according to the type of hydrophobic thin film and the metal thin film.

In addition, the high sensitivity array chip according to the second embodiment of the present invention shown in FIG.

(S1) depositing a metal thin film on the substrate;

(S2) depositing a hydrophobic thin film on the metal thin film;

(S3) patterning the metal thin film and the hydrophobic thin film through a photolithography process;

(S4) forming a plurality of microwells by etching the metal thin film and the hydrophobic thin film on which the pattern is formed; And

(S5) Produced by forming a hydrophilic thin film having a hydrophilic functional group on the surface of the substrate inside the opening of the microwell.

At this time, the steps of S1 to S4 are as described above.

The highly sensitive array chip according to the second embodiment is manufactured by forming a hydrophilic thin film on the surface of the substrate inside the opening of the microwell after the step S4 and modifying the surface thereof.

Specifically, as shown in FIG. 9, a hydrophilic thin film 9 having a hydrophilic functional group is formed on the surface of the substrate 1 in the opening region of the microwell 3 (S5).

The hydrophilic thin film 9 is formed using a self-assembled thin film method using a solution containing one hydrophilic group selected from the group consisting of a hydroxyl group, an amine group, an aldehyde group, a carboxyl group and a combination thereof.

If necessary, the hydrophilic thin film 9 may be further modified to have a specific hydrophilic group to form a modified hydrophilic thin film 9a (FIG. 10, where R = OH, NH ₂ , COH, or COOH).

The modified hydrophilic thin film 9a includes a specific hydrophilic group required in the field of using an array chip. The modified hydrophilic thin film 9a is formed by dropping or spraying a compound containing a specific hydrophilic group onto the hydrophilic thin film 9 to react with the hydrophilic group of the previously formed hydrophilic thin film 9.

The high-sensitivity array chip manufactured through such a step has various fields such as IT (Information Technology), BT (Bio Technology), ET (Environment Technology), NT (nano technology), AT (Agriculture Technology) or fusion fields thereof. Applicable to the field. For example, the high sensitivity array chip can be used for analyzing biomolecules such as proteins and nucleic acids.

In the highly sensitive array chip according to the present invention, the lower part of the microwell opening into which the sample is injected is modified with a hydrophilic functional group so that the sample can be selectively fixed more easily.

As shown in FIGS. 11 and 12 (a) and (b), the microwells are made of a hydrophobic thin film so that the sample solution does not get wet around the microwells, and can be aligned and concentrated only inside the microwells, thus providing 0.1 to 1.0 μl. Analysis is possible even with very small samples of / spot.

Particularly, due to the metal thin film, the optical signal can be selectively suppressed to obtain a very good signal-to-noise ratio compared to the conventionally manufactured array chip, and the sample is aligned in the exact arrangement of the opening in which the hydrophilic thin film is formed in advance. In addition, since the sample is hardly adsorbed to the hydrophobic thin film portion, more reliable analysis results can be obtained.

In addition, the hydrophobic / hydrophilic surface of the high-sensitivity array chip can be measured in the same manner as in the case of using a conventional planar chip even when the entire chip is analyzed by maintaining a plan view of less than a micrometer unit.

In addition, no expensive arrangement is required, and the biomaterial can be easily immobilized in a desired pattern, making it possible to manufacture a user-friendly platform.

Hereinafter, preferred examples and experimental examples of the present invention are presented. However, the following examples are only preferable examples of the present invention, and the present invention is not limited to these examples.

Experimental Example 1

In order to confirm the wettability of the sample in the microwell, the contact angle of the hydrophilic thin film and the hydrophobic thin film on the substrate was measured.

First, a 100 nm thick Ti thin film is deposited on a silicon substrate, and then a self-assembled thin film is formed with an aminopropyltriethoxysilane to a thickness of 2 nm on top of the silicon substrate / Ti thin film / hydrophilic self-assembled thin film. Formed.

Alternatively, the self-assembled thin film was formed to have a thickness of 2 nm on the Ti thin film by using FOTS (fluorooctyltrichlorosilane), thereby forming a silicon substrate / Ti thin film / hydrophobic self-assembled thin film.

(A) of FIG. 13 is a contact angle of a Ti thin film formed on a silicon substrate, (b) shows a hydrophilic self-assembled thin film having an amine group on its surface, and (c) shows a contact angle of a hydrophobic self-assembled thin film.

Referring to FIG. 13, the Ti contact angle was 47 °, the hydrophobic self-assembled thin film was 100 kV, and the hydrophilic self-assembled thin film was 40 °. As a result, when the lower part of the microwell into which the sample is injected has hydrophilicity and the surroundings are treated hydrophobicly, the hydrophilic thin film may result in higher wettability of the sample to the well, and due to the repulsive force with the hydrophobic thin film. It can be seen that the sample can only be effectively present inside the well.

Experimental Example 2

Ti metal was deposited by sputtering on a silicon substrate (thickness of 100 nm), and a self-assembled thin film (thickness of 2 nm) was formed using FOTS (fluorooctyltrichlorosilane) thereon. After applying PR (AZ 1518) on the self-assembled thin film and performing exposure and development, the patterned multilayer thin film was obtained by patterning.

The self-assembled thin film was formed to a thickness of 2 nm with aminopropyltriethoxysilane on the silicon substrate on which the multilayer thin film was not formed, and then, by dissolving glutaraldehyde, the surface was modified with aldehyde to form an microwell array. Prepared.

The arrays were selectively immobilized on 4 × 4 arrays using BSA proteins (CY5-BSA) labeled with CY5 fluorescent chromosomes.

FIG. 14 is an image of fluorescent scan after introducing a fluorescent labeled protein measured after modifying a substrate with an aldehyde group. Referring to FIG. 14, it can be seen that the protein is selected and fixed with high sensitivity only to the hydrophilic selection part.

Experimental Example 3

Ti metal was deposited by sputtering on a silicon substrate (thickness of 100 nm), and a self-assembled thin film (thickness of 2 nm) was formed using FOTS thereon. After applying PR (AZ1518) on the self-assembled thin film, exposure and development were performed, and then patterned to obtain a patterned multilayer thin film.

A self-assembled thin film was formed to a thickness of 2 nm with aminopropyltriethoxysilane on the silicon substrate on which the multilayer thin film was not formed, and then, acetic acid was dispensed to modify the surface with a carboxyl group to prepare an array in which microwells were formed.

The array was selectively immobilized to the 4 × 4 array using BSA protein (CY3-BSA) labeled with CY3 fluorescent chromosome.

FIG. 15 is an image of fluorescent scan after introduction of a fluorescent labeled protein measured after modifying a substrate with an aldehyde group. Referring to Figure 15, it can be seen that the protein is fixed to a selective and high sensitivity only to the hydrophilic select portion.

The highly sensitive array chip according to the present invention can be used for analyzing biomolecules such as proteins and nucleic acids.

1 and 2 are a cross-sectional view and a three-dimensional view showing a high sensitivity array chip according to a first embodiment of the present invention.

3 is a cross-sectional view and a three-dimensional view showing a high sensitivity array chip according to a second embodiment of the present invention.

4 to 10 are cross-sectional views illustrating a manufacturing procedure of the high sensitivity array chip according to the present invention.

11 and 12 are schematic diagrams showing the application of the high sensitivity array chip according to the present invention.

(A) of FIG. 13 is a contact angle of a Ti thin film formed on a silicon substrate, (b) shows a hydrophilic self-assembled thin film having an amine group on its surface, and (c) shows a contact angle of a hydrophobic self-assembled thin film.

14 is a fluorescence scan image measured after modifying a substrate with an aldehyde.

15 is a fluorescence scan image measured after modifying a substrate with a carboxyl group.

Claims (19)

Having a plurality of microwells on a substrate; The microwell is a high sensitivity array chip formed of a multilayer thin film in which a metal thin film and a hydrophobic thin film are stacked. Having a plurality of microwells on a substrate; The microwell is formed of a multilayer thin film in which a metal thin film and a hydrophobic thin film are stacked. And a hydrophilic thin film having a hydrophilic functional group formed on a substrate inside the opening of the microwell. The method according to claim 1 or 2, The substrate is a high sensitivity array chip comprising one selected from the group consisting of glass, semiconductor and plastic. The method according to claim 1 or 2, The metal thin film is a high sensitivity array chip comprising one selected from the group consisting of Ti, Cr, Al, Cu, Au, Ag, Pt, Si, Rh, Ru, V, Ta and combinations thereof. The method according to claim 1 or 2, The hydrophobic thin film is perfluoro trichlorosilane (FOTS, Perfloroctyltrichlorosilane), octadecylmercaptan (ODT, Octadecylmercaptan), trichlorooctadecylsilane (OTS, Trichlorooctadecylsilane), dichlorodimethylsilane (DDMS, Dichlorodimethylsilane), undecyl Trichlorosilane (DTS, Undecyltrichlorosilane), Undecenyltrichlorosilane (Undecenyltrichlorosilane), Perfluorodecyltrichlorosilane (PDTS, Perfluorodecyltrichlorosilane), Trichlorovinylsilane (TVS, Trichlorovinylsilane), Perfluorodecanoic acid (PFDA , Perfluorodecanoic Acid) and a high sensitivity array chip that is a self-assembled thin film (Self-Aligned Monolayer) comprising one selected from the group consisting of these. The method according to claim 1 or 2, The hydrophobic thin film is selected from the group consisting of source gas such as CHF ₃ , CH ₂ F ₂ , C ₂ F ₆ , C ₃ F ₈ , C ₄ F ₈ , C ₅ F ₈ , C ₄ F _6, and combinations thereof. A highly sensitive array chip comprising a hydrophobic thin film deposited by a plasma enhanced chemical vapor deposition method including a species. The method according to claim 1 or 2, The hydrophobic thin film may be a polyvinylphenol polymer, a polyhydroxystyrene polymer, a polynorbornene polymer, a polyadamant polymer, a polyimide polymer, a polyacrylate polymer, a polymethacrylate polymer, or a polyfluorine polymer. High sensitivity array chip comprising a photoresist selected from the group consisting of polymers and combinations thereof. The method according to claim 1 or 2, The hydrophobic thin film is one selected from the group consisting of polytetrafluoroethylene (PTFE), polychlorotrifluoroethylene (PCTFE), polyvinylidene fluoride (PVDF), polyvinyl fluoride (PVF), and combinations thereof High sensitivity array chip comprising a fluororesin (Fluororescein) comprising a. The method according to claim 1 or 2, The hydrophobic thin film is 2-tert-butyl-9,10-di (2-naphthyl) anthracene, anthracene, 9,10-diphenylanthracene, tetracene, rubin, perylene, parylene N, Parylene C, Parylene CF, Parylene D, Parylene HT, Parylene SR, Parylene SF, 2, A highly sensitive array chip comprising one aromatic hydrocarbon selected from the group consisting of 5,8,11-tetra (tert-butyl) perylene, pentacene, coronine, and combinations thereof. The method according to claim 1 or 2, The hydrophobic thin film is a high sensitivity array chip comprising one selected from the group consisting of polydimethylsiloxane, linear polyimides (Linear polyimides), aromatic heterocyclic polyimides (Aromatic heterocyclic polyimides) and combinations thereof. The method according to claim 1 or 2, The microwell is a highly sensitive array chip that is cylindrical, polyhedron, inverted cone, or inverted pyramidal shape. The method of claim 2, Wherein the hydrophilic functional group is a highly sensitive array chip that comprises one selected from the group consisting of hydroxy group, amine group, aldehyde group, carboxyl group, and combinations thereof. The method of claim 2, The hydrophilic thin film is R ^1- (CH ₂ ) n -Si- (R ² ) ₃ , wherein R ¹ = OH, NH ₂ , COH, or COOH, R ² is Cl or OC _n H _{2n + 2} alkoxy A high sensitivity array chip which is trichlorosilane or trialkoxysilane represented by (alkoxy)). Depositing a metal thin film on the substrate; Depositing a hydrophobic thin film on the metal thin film; Patterning the metal thin film and the hydrophobic thin film through a photolithography process; And Forming a plurality of microwells by etching the patterned metal thin film and the hydrophobic thin film. Depositing a metal thin film on the substrate; Depositing a hydrophobic thin film on the metal thin film; Patterning the metal thin film and the hydrophobic thin film through a photolithography process; Etching the patterned metal thin film and the hydrophobic thin film to form a plurality of microwells; And Forming a hydrophilic thin film having a hydrophilic functional group on a substrate surface inside the opening of the microwell Method of manufacturing a high sensitivity array chip comprising a. The method according to claim 14 or 15, The metal thin film deposition is performed by one method selected from sputtering, ion beam deposition, electron beam deposition, thermal deposition, laser molecular beam deposition, pulsed laser deposition, metal organic chemical vapor deposition, chemical vapor deposition, and plasma deposition. Manufacturing method. The method according to claim 14 or 15, The deposition of the hydrophobic thin film is a method of manufacturing a highly sensitive array chip that is carried out by a self-assembled thin film formation method, plasma enhanced chemical vapor deposition method or wet coating method. The method of claim 15, The hydrophilic thin film is a method of manufacturing a high sensitivity array chip is formed by a self-assembled thin film formation method. The method of claim 15, The hydrophilic thin film is a method for producing a highly sensitive array chip that further modify the surface of the hydrophilic functional group.

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