KR101460066B1 - Fabricating method for nanowell array biosensor on a large sized substrate - Google Patents

Abstract

The present invention relates to a nanowell array biosensor in a large area substrate and a method of manufacturing the same, and more particularly, to a nanowell array biosensor having a nanowire array structure reproducibly realized by a novel method, 0001] The present invention relates to a nanowell array biosensor in a large-area substrate and a method of manufacturing the same.
According to the present invention, a nano-sized well array structure highly integrated in a narrow region can be realized uniformly and reproducibly. Accordingly, it is possible to manufacture a biosensor having greatly improved detection sensitivity, selectivity, and reliability of biomolecules such as various enzymes, proteins, DNA, and biomaterials. Also, according to the present invention, large-area substrates having a size of 6 inches (150 mm in diameter) or larger can be mass-produced unlike the prior art, and the possibility of commercialization is high.

Description

TECHNICAL FIELD [0001] The present invention relates to a biosensor array for biosensor array,

The present invention relates to a method of fabricating a nanowell array biosensor in a large-area substrate, and more particularly, to a nanowell array biosensor with a novel method to reproducibly realize a nano-sized well array structure to improve detection sensitivity, selectivity and reliability To a method of manufacturing a nanowell array biosensor on a large area substrate.

Recently, research on biosensors capable of quickly and accurately measuring biomolecules, biomaterials, and the like at any time and anywhere has been actively conducted due to an increased interest in medical care, the environment, food, and military. Medical biosensors are expected to play a leading role in the future growth of the biosensor industry as well as the present. Currently, medical sensors account for 11% of the total sensor market, including 49% of chemical sensors, 25% of ultrasonic sensors, 9% of biosensors, 7% of pressure sensors, 6% of temperature sensors and other sensors. Biosensors are used in 85-90% for medical use, and blood glucose sensors account for 90% in medical use.

Among various types of biosensors, electrochemical sensors are receiving the most attention because they are accurate, fast and economical, are easy to miniaturize, and can measure a wide range of signals (currents) with a simple measuring device. Combining the basic advantages of such an electrochemical sensor with a detection technique that provides quick detection and a low detection limit, a practical biochemical biosensor can be realized. In recent years, nanoscale electrochemical biosensors have been reduced in size to nanoscale, enabling analytical ability, local concentration measurement, and dopamine secretion to be measured, enabling in vivo monitoring of neurochemical applications.

However, it still has many limitations. (KR) Patent No. 10-1092859 provides a method for manufacturing microcontact printing, which is very low in reproducibility because it utilizes the movement of bio-content due to physical contact, and is very disadvantageous in large-area pattern formation Do.

The manufacturing method of a nanobio array using the SPM mentioned in Korean Patent (KR) No. 10-0506508 takes a very long time to form a large-area uniform nano array, and in fact, the formation of a large-area nano pattern is limited.

The method of manufacturing a nanocylindrical template or nano-dot array described in Korean Patent (KR) No. 10-1033806 has proposed a relatively economical and simple manufacturing method as compared with existing manufacturing techniques, There is a limit to the formation of the pattern, and it is difficult to control the size of the nanopattern as compared with the top down process, and the reproducibility and reliability of the continuously produced nanopattern are low.

Korea (KR) Patent No. 10-0549104 discloses a method of producing a nanopattern having a concave or columnar shape of less than several nanometers by using self-assembly of organic molecules and selective chemical adsorption of metals, but the nanopattern is not uniform and very irregular There is a problem that it is difficult to apply a large area. Korean Unexamined Patent Publication No. 10-2012-0060968 discloses a high density uniform nanoparticle array using a mixed block copolymer in which a high molecular weight first polymer and a low molecular weight second polymer are mixed. However, There are limitations in controlling various shapes and sizes of nanopatterns, and there are limitations in view of reproducibility, reliability, and mass production methods in view of a large area.

Korean Patent No. 10-1033806 Korean Patent No. 10-0506508

The present invention provides a nano-well array biosensor in a large-area substrate and a method of manufacturing the same, which can improve detection sensitivity, selectivity and reliability of a biomaterial by realizing a nano-sized well array structure on a large-area substrate in a reproducible manner .

A method of fabricating a nanowell array biosensor in a large area substrate,

Forming an oxide film on the substrate; Depositing a metal layer on the oxide layer; Forming a first photosensitive layer by coating a photosensitive agent on the metal layer, and forming a fine pattern through a photolithography process using a stepper exposure equipment; Etching the metal layer in the fine pattern shape, removing the photosensitive layer, and depositing an insulating layer; Forming a second photosensitive layer by coating a photosensitive agent on the insulating layer, and forming a nano pattern through a photolithography process using a stepper exposure equipment; And a second step of etching the insulating layer in the nanopatterned shape.

The substrate may be of a large area with a diameter of at least 6 inches and the substrate may be a silicon substrate.

Oxide film  Forming step

The oxide film formed on the substrate may be a silicon oxide film formed of silicon.

The oxide layer may be formed by a plasma chemical vapor deposition method or a thermal oxidation method, and the oxide layer may be formed to a thickness of 1000 to 5000 ANGSTROM.

Metal layer Deposition step

The metal layer may be formed of at least one metal selected from the group consisting of gold, platinum, silver, palladium, copper, and nickel for the electrode structure of the biosensor. However, the present invention is not limited thereto, and any conductive material used for electrochemical analysis can be used. However, it is preferable to use gold or platinum when the biosensor of the nano-well array structure is completed and the surface treatment for biomaterial treatment is important.

The metal layer may be formed by a conventional deposition method such as sputtering, evaporation, or ion beam deposition.

Fine pattern formation step

The first photosensitive layer is formed by applying a photosensitive agent to the deposited metal layer. The first photosensitive layer may have a thickness ranging from 0.5 to 1 μm. The thickness of the first photosensitive layer is preferably determined in consideration of etch selectivity between the first photosensitive layer and the metal layer in the step of etching the metal layer after the fine pattern formation step. When the thickness of the first photosensitive layer is too small (less than 0.5 μm), the photosensitive layer may be blown off before the metal layer is completely etched during the plasma dry etching process, and the desired fine pattern can not be formed. Further, if the thickness of the first photosensitive layer is excessively large (exceeding 1 占 퐉), it may not be easy to remove the first photosensitive layer after the plasma dry etching process.

A photoresist may be applied to the metal layer to form a first photosensitive layer, and then a fine pattern may be formed on the first photosensitive layer through a photolithography process using a stepper exposure equipment.

The stepper exposure equipment may be a KrF stepper exposure equipment or an ArF stepper exposure equipment. In addition to the stepper exposure apparatus, it is possible to use a more economical exposure apparatus. However, in the process of forming a nano-well array according to the present invention, alignment of first and second patterns (fine patterns and nanopatterns) It is desirable to form a fine pattern with a stepper exposure equipment. The stepper exposure apparatus can form a pattern by repeatedly step-projecting ultraviolet light emitted through a reticle, which is a circuit disk having fine patterns, onto a wafer substrate through a lens. In this case, a KrF stepper exposure apparatus using an excimer laser having a wavelength of 248 nm and an ArF stepper exposure apparatus using an excimer laser having a wavelength of 193 nm can be applied. Here, the reticle is a mask to be used for lithography, and is the first mask to finally form the nano-well array structure of the present invention. The reticle is a pattern to be formed on the photosensitive layer, and a shape conforming to the finally formed nano- well array biosensor Characterized by comprising a fine pattern. At this time, a highly uniform and reproducible nano-well array pattern can be formed on a large area substrate by applying a reticle having various patterns according to the shape, arrangement, and size of the required nano-well array structure. After the exposure, the photosensitive layer may be selectively removed by development to form a fine pattern.

The fine pattern size may be from a few tens of micrometers to several millimeters. At this time, the size of the fine pattern is preferably large enough to be fixed to the equipment or cell for electrochemical analysis, and can be adjusted according to the analysis equipment and analysis method.

Metal layer Etching step

The metal layer may be first etched using the photosensitive layer having the fine pattern formed thereon as a mask to form the fine pattern on the metal layer. Accordingly, the metal layer may have the same fine pattern as the fine pattern formed on the first photosensitive layer. The first etching may be dry etching. The dry etching is preferably performed by Reactive Ion Etching (RIE) or Inductively Coupled Plasma (ICP) etching.

The reactive ion etching or inductively coupled plasma etching is preferably performed under the conditions of a Cl2 gas at 25 to 50 sccm, an Ar gas at 10 to 20 sccm, a pressure of 100 to 200 mTorr, and a plasma power of 50 to 300 W.

When the first etching is finished, the first photosensitive layer may be removed. At this time, the photosensitive layer may be dipped in acetone and then removed using ultrasonic.

Insulating layer  Forming step

The insulating layer may be formed of any material that is not electrically conductive, but a silicon dioxide (SiO ₂ ) or a silicon nitride (SiN x) film having excellent adhesion to the substrate and the metal layer and being easy to etch is used desirable. The silicon oxide film and the silicon nitride film are the most widely used materials for forming the insulating layer in the semiconductor and display industries. The adhesion of the silicon oxide film and the silicon nitride film to the base material such as silicon and metal has been sufficiently verified. In the etching process after the deposition, Because of its high selectivity, it is easy to apply and has biocompatibility. Therefore, it can be used easily as a biosensor after chip fabrication.

Since the depth of the nano-well structure is determined in the nano-well array biosensor in which the thickness of the insulating layer to be deposited is finally produced, the thickness of the insulating layer may be important. A preferable thickness of the insulating layer may be 100 to 9000 ANGSTROM. If the thickness of the insulating layer is less than 100 angstroms, the nonspecific binding of the biomaterial having a relatively larger size as compared to the depth (height) of the nano-well may be increased to increase noise, As the depth (height) of the nano-well is deeper than the diameter of the nano-pattern, electrons and ions are trapped inside the nano-well, which may hinder the sensitivity and selectivity of the sensor. Is preferably in the above range.

The insulating layer may be deposited using plasma enhanced chemical vapor deposition.

In the case of depositing a silicon oxide film (SiO ₂ ) as the insulating layer, a silicon oxide film (SiO ₂ ) is deposited by plasma CVD (PECVD) using 100 to 200 sccm of 5% SiH ₄ / He gas, 500 to 800 sccm of N ₂ O gas, 300 ° C, 500-1000 mTorr pressure, and 50-150 W plasma power.

Also, when depositing a silicon nitride film (SiNx) with the insulating layer, 5% by using a plasma enhanced chemical vapor deposition (PECVD) SiH ₄ / He gas 300 ~ 450 sccm, NH ₃ The deposition is preferably performed under the conditions of 10 to 50 sccm of gas, 600 sccm of N ₂ gas, a deposition temperature of 100 to 300 ° C, a pressure of 1500 to 2000 mTorr, and a plasma power of 20 to 50 W.

Nano-pattern formation step

The second photosensitive layer is formed by applying a photosensitive agent to the deposited insulating layer. The second photosensitive layer may have a thickness ranging from 0.5 to 1 μm. The thickness of the second photosensitive layer is preferably determined in consideration of the etch selectivity between the second photosensitive layer and the insulating layer in the step of etching the insulating layer after the nano pattern forming step. If the thickness of the second photosensitive layer is too small (less than 0.5 탆), the photosensitive layer may be blown off before the insulating layer is completely etched during the plasma dry etching process, and the desired fine pattern can not be formed . Further, if the thickness of the second photosensitive layer is too large (exceeding 1 占 퐉), it may not be easy to remove the second photosensitive layer after the plasma dry etching process.

The photosensitive layer may be formed on the deposited insulating layer to form a second photosensitive layer, and then a nano pattern may be formed on the second photosensitive layer using a photolithography process using a stepper exposure equipment. In the present invention, a nano-pattern means a nano-sized pattern.

In the process of forming a nano pattern by forming a photolithography process by a stepper exposure equipment, ultraviolet light emitted through a reticle having a nano pattern is repeatedly projected onto a large-area substrate through a lens in the same manner as the above- (step projection). At this time, since the nano-well array structure should be formed on the fine pattern of the pre-formed metal layer, alignment must be accurately performed using an align key existing on the reticle. A nanopattern may be formed on the second photosensitive layer formed on the insulating layer by development after exposure.

The nanopattern may be at least one selected from the group consisting of a circular array, a polygonal array having a square or more and a honeycomb array, and the array of the nanopatterns may be formed in a parallel array or a zigzag array. But it is not limited thereto and may be patterned and arranged in a known shape.

The nanopattern may be formed with a diameter and an interval of 100 to 1000 nm. When the diameter and the interval of the nano patterns are less than 100 nm, it is impossible to form a pattern because the pattern resolution of the KrF stepper exposure apparatus or the ArF stepper exposure apparatus deviates from the pattern resolution. When the diameter exceeds 1000 nm (1 μm) , Detection sensitivity, selectivity, reliability, etc. of the nano-well structure can not be derived. Therefore, the above range is preferable.

Insulating layer Etching step

And the second layer of the insulating layer is etched using the second photosensitive layer having the nano pattern formed thereon as a mask to form a nano pattern on the insulating layer. Accordingly, the same nano pattern as the nano pattern formed on the second photosensitive layer may be formed on the insulating layer.

The second etching may be dry etching. The etching of the insulating layer is preferably performed by reactive ion etching (RIE) or inductively coupled plasma (ICP) etching as in the first etching (metal layer etching) step.

The reactive ion etch or inductively coupled plasma etch may be performed using CF ₄ 30 to 50 sccm of gas, 10 to 500 mTorr of pressure, and 30 to 300 W of plasma power.

The second photosensitive layer removing step

The method of fabricating a nanowell array biosensor in the large area substrate may further include removing the second photosensitive layer after the second etching of the insulating layer. At this time, the photosensitive layer may be dipped in acetone and then removed using ultrasonic.

According to the present invention, there is provided a nano-well array biosensor in a large-area substrate manufactured by the above-described method, wherein the nanowell array in a large-area substrate used for environmental bio-monitoring, medical diagnosis, food toxicity evaluation, A biosensor can be provided. (See Fig. 1)

The nanowell array biosensor in a large-area substrate according to the present invention can be used not only for environmental bio-monitoring, medical diagnosis, food toxicity evaluation or cosmetic toxicity evaluation, but also immunosensor, gene expression analysis, Such as enzymes, DNA, RNA, and sensors for protein detection. It is expected to be applied to various fields such as information, environment, medical and military fields in the future. More specifically, the present invention relates to a method for monitoring atmospheric and water quality, a nano-biosensor for animals and plants, a new drug test, an early diagnosis of crops and animal diseases, an early diagnosis and prediction of cancer outbreak, a food characteristic and toxicity detection, And can be utilized as a nano-biosensor.

According to the present invention, a nano-sized well array structure highly integrated in a narrow region can be realized uniformly and reproducibly. Accordingly, it is possible to manufacture a biosensor having greatly improved detection sensitivity, selectivity, and reliability of biomolecules such as various enzymes, proteins, DNA, and biomaterials. Also, according to the present invention, large-area substrates having a size of 6 inches (150 mm in diameter) or larger can be mass-produced unlike the prior art, and the possibility of commercialization is high.

1 is a schematic perspective view of a nano-well array biosensor in a large-area substrate.
2 is a flowchart of a method of manufacturing a nano-well array biosensor in a large-area substrate.
3 is a flow chart of a method of manufacturing a nano-well array biosensor on a large-area substrate.
4 is a photograph of (a) 6 inch wafer sample, (b) one nanowell array biosensor according to Example 1, and (c) 57 chips made in one 6 inch.
5 shows the (ac) SEM image of the nano-well array structure according to Example 1 and (d) the EDS analysis result of the surface.
6 is an (ac) AFM image and (de) current distribution image for the nano-well array structure according to Example 1. Fig.
7 is a schematic diagram of a biomaterial (STIP 1) treatment in a nano-well array biosensor according to Example 1 of the present invention.
FIG. 8 shows the results of (a) electrochemical impedance analysis (ElS) and (b) cyclic voltammetry (CV) measurements of Example 1 and Comparative Example 1.
FIG. 9 shows electrochemical impedance analysis (EIS) results of Example 1 (a) and Comparative Example 1 (b) according to STIP 1 concentration.
10 is a standard curve showing electrochemical analysis (EIS) results according to STIP 1 concentration.

Hereinafter, the present invention will be described in more detail with reference to Examples. The objects, features and advantages of the present invention will be readily understood by the following examples. The present invention is not limited to the embodiments described herein but may be embodied in different forms. Therefore, the present invention should not be limited by the following examples.

Example 1

Fabrication of nanowell array biosensors on large area substrates (see Figs. 2 and 3)

A P-type (100) silicon substrate having a size of 6 inches (150 mm in diameter) was prepared and a 3000 Å thick silicon oxide film was formed by thermal oxidation. Next, a metal layer having a thickness of 3000 Å was deposited on the silicon oxide film from a gold (Au) target using a sputtering apparatus. Thereafter, a photosensitive agent was coated on the metal layer, and then a fine pattern was formed on the photosensitive layer by a reticle having an electrode pattern of 8 mm ² size formed by using a KrF stepper exposure equipment. Thereafter, using a photosensitive layer having a fine pattern formed thereon as a mask, a 3000 Å thick gold (Au) layer was formed in an ICP etching apparatus under the conditions of Cl 2 gas of 35 sccm, Ar gas of 10 sccm, pressure of 120 mTorr, Au) metal layer was selectively removed, and then the photoresist was removed by immersing in an ultrasonic beaker for 3 minutes using acetone (step d). Next, 150 sccm of a 5% SiH ₄ / He gas, 700 sccm of N ₂ O gas, a temperature of 100 ° C, a pressure of 1000 mTorr (10 mTorr) using a plasma chemical vapor deposition (PECVD) equipment for forming an insulating layer constituting the nano- , then under the conditions of plasma power 120 W deposit a silicon oxide film (SiO ₂₎ of 2000Å thickness was coated with a photosensitive material thereon in a thickness of 7000Å (step e). Next, to form a nano-well array pattern having a regular arrangement in the photosensitive layer, a nano pattern formed by a reticle in which a regular circular array pattern having a diameter and a pattern interval of 400 nm were formed using a KrF stepper exposure equipment (Step f). Then, a silicon oxide film (SiO ₂ ), which is an insulating layer, was selectively formed in a reactive ion etching (RIE) equipment using a photosensitive layer having a nano pattern formed thereon as a mask under conditions of CF 4 gas 30 sccm, pressure 150 mTorr, (Step g). Thereafter, the photosensitizer was immersed in an ultrasonic beaker for 3 minutes using acetone (step h) to finally produce a nanowell array biosensor in a large-area substrate according to the present invention.

Comparative Example 1

A P-type (100) silicon substrate having a size of 6 inches (150 mm in diameter) was prepared as a reference for a nano-well array biosensor according to Example 1, a silicon oxide film having a thickness of 3000 Å was formed by thermal oxidation, A bare Au electrode was fabricated by depositing a metal layer having a thickness of 3000 Å on the silicon oxide film from a gold (Au) target using a sputtering apparatus.

Experimental Example 1

Photo image analysis

FIG. 4A shows a 6-inch wafer sample manufactured according to Example 1, FIG. 4B shows an enlarged photograph of one chip, and FIG. 4C shows photographs of 57 chips manufactured on one 6-inch wafer. Respectively.

Experimental Example 2

SEM image analysis

FIG. 5 shows a SEM image of the nano-well array structure analyzed by a scanning electron microscope (SEM) in the nano-well array biosensor manufactured according to Example 1. FIG. FIG. 5A shows the nano-well array structure very clearly as a SEM image measured at an inclination of 45 °. FIG. 5B is a SEM image of a cross section of the nano-well array structure, which shows the diameter, spacing, and depth of the nano-well. FIG. 5C can clearly confirm that the nano-well pattern is formed in a very uniform manner as a SEM image obtained by measuring a slightly wider area. FIG. 5D shows that the surface of the nano-well array structure is analyzed by energy-dispersive X-ray spectroscopy (EDS) and that a gold (Au) metal layer and a silicon oxide film (SiO ₂ ) are present.

Experimental Example 3

AFM analysis

The nano-well array structure fabricated according to Example 1 was analyzed by AFM (Atomic Force Microscope) and the results are shown in FIG. 6 (a) to (c), the surface topology of the nanoscale can be confirmed more precisely. 6D and 6E show the results of measurement using the I-AFM (current sensing AFM) mode. It can be confirmed that the current is uniformly distributed in the nano-well array structure according to the first embodiment. This current distribution confirmation is very important as a method for surely proving that the nano-well array biosensor manufactured according to the first embodiment is properly manufactured as a non-defective good product.

Experimental Example 4

In order to confirm the actual performance of the nano-well array biosensor manufactured according to Example 1, sensitivity, selectivity, and reliability as a biosensor using a protein as a biomaterial were examined. Experiments were conducted. The biomaterial is a protein that occurs in the body when stress induced STIP 1 (Stress-Induced-Phosphoprotein-1), which is used as a target detection substance. Biotest experiments were carried out using the binding reaction between streptavidin and biotin, which is commonly known for detecting STIP 1 in the nano-well array region. A liquid self assembled monolayer was used for the surface treatment in the nano well. The surface treatment by the liquid phase self-assembled thin film method was carried out by mixing 11-mercaptoecanoic acid (11-MUA) at a concentration of 10 mM in anhydrous ethanol, immersing the nanolow array sample at room temperature for 1 hour, Was treated with a solution of 50 mM EDC and 50 mM NHS in a sodium acetate buffer (pH 5.5) for 1 minute to activate the ester surface. Next, in order to immobilize streptavidin on the electrode surface in the nano-well, the sample was immersed in PBS buffer containing 1 mg / ml of streptavidin for 30 minutes at room temperature. Then, biotinylated antibody (10 μg / ml) was treated with a nano-well array biosensor for 1 minute and electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) The detection sensitivity and pattern of STIP 1 antigen were measured according to the concentration of STIP 1 antigen. (Refer to FIG. 7). At this time, an oxide film was formed on a silicon substrate with a reference of a nano-well array biosensor, and then a pure Au bare Au electrode Example 1) together with the biomaterial treatment.

Experimental Example 4-1

Analysis by Electrochemical Impedance Analysis (EIS) and Cyclic Voltammetry (CV)

The electrochemical impedance analysis (EIS) results of the comparison between the nano-well array biosensor manufactured according to Example 1 and the pure gold electrode prepared according to Comparative Example 1 are shown in FIG. 8A. The measurement results are shown in Fig. 8B.

As shown in FIG. 8A, the charge transfer resistance (Rct) values of the nano-well array according to Example 1 and the pure gold electrode according to Comparative Example 1 were 20.87 kohm and 42.98 kohm, respectively. The lower resistance values in the nano-well array structure indicate that more current can flow in the nano-well array structure. This is because, in the conventional large electrode, the movement of the material linearly diffuses, whereas in the nano-sized well structure, concentration occurs in the nanowire in the electrical double layer, resulting in radial diffusion, Can be interpreted as facilitating.

As shown in FIG. 8B, the total charge amount Q of the nano-well array structure is 27.05 mC, which is higher than the total charge amount 14.88 mC of the pure gold electrode. This indicates that more current can flow in the nano-well array structure, which supports the result of FIG. 8A.

Experimental Example 4-2

Electrochemical Impedance Analysis by STIP 1 Concentration

The electrochemical impedance analysis (EIS) results of the nano-well array biosensor according to Example 1 of the present invention are shown in FIG. 9A, and the electrochemical impedance analysis (EIS) results of the pure gold electrode according to Comparative Example 1 are shown in FIG. 9B . As the concentration of STIP 1 antigen increased in the nano-well array structure, the charge transfer resistance (Rct) was also increased, and also showed a similar increase in the pure gold electrode. However, as shown in FIG. 9A, the difference in charge transfer resistance according to concentration is clearly distinguished in the nano-well array structure. However, as shown in FIG. 9B, the difference in charge transfer resistance between pure gold electrodes is very small.

In order to be used as a biosensor, the detection signal according to each concentration must show a distinct difference, which is related to the selectivity among the requirements of the sensor. As described above, in the nano-well array structure according to the first embodiment, the selectivity of the requirements of the sensor is very excellent because the difference in the charge transfer resistance according to the concentration is clearly distinguished.

Experimental Example 4-3

Analysis according to standard curve

The results of the electrochemical impedance analysis according to the concentration of STIP 1 in Experimental Example 4-2 were calculated and shown in FIG. 10 as standard curves. Pure gold (Bare Au) on the amount of change (ΔR _ct) This showed a larger, pure gold (Bare Au) STIP 1 antigen concentration in nano-well array structure than the electrodes of the charge transfer resistance (Rct) in the nano-well array structure than that of electrode The value of which is clearly distinguished. From the viewpoint of the most important detection limit of the sensor, the nano-well array structure can be detected even at a very low antigen concentration of 10 pg / ml. Therefore, it can be considered that the biosensor is highly utilized as a biosensor. It is interpreted that the nano-well structure is also advantageous for electrical signal detection, but it also influences the lowering of the probability of nonspecific binding occurring in the processing of biomaterials.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the invention. For example, the nano-well array biosensor described in the embodiments of the present invention exemplifies the use of a KrF stepper exposure apparatus and the application of a reticle having a regular circular array pattern having a pattern diameter and an interval of 400 nm, respectively, ArF stepper exposure equipment and various types of reticles and the like can be applied within the technical scope of the present invention.

Claims (16)

Forming an oxide film on the substrate;
Depositing a metal layer on the oxide layer;
Forming a first photosensitive layer by coating a photosensitive agent on the metal layer, and forming a fine pattern through a photolithography process using a stepper exposure equipment;
Etching the metal layer in the fine pattern shape, removing the first photosensitive layer, and depositing an insulating layer;
Forming a second photosensitive layer by coating a photosensitive agent on the insulating layer, and forming a nano pattern through a photolithography process using a stepper exposure equipment; And
And etching the insulating layer in a nanopatterned shape. ≪ RTI ID = 0.0 > 21. < / RTI > The method according to claim 1,
Wherein the substrate is at least 6 inches in diameter. ≪ Desc / Clms Page number 20 > The method according to claim 1,
Wherein the substrate is a silicon substrate. ≪ RTI ID = 0.0 > 8. < / RTI > The method according to claim 1,
Wherein the oxide film is a silicon oxide film formed of silicon. The method according to claim 1,
Wherein the oxide film is formed by a plasma chemical vapor deposition method or a thermal oxidation method. The method according to claim 1,
Wherein the metal layer is made of at least one metal selected from the group consisting of gold, platinum, silver, palladium, copper, and nickel. The method according to claim 1,
Wherein the first photosensitive layer and the second photosensitive layer are each formed to a thickness ranging from 0.5 to 1 占 퐉. The method according to claim 1,
Wherein the stepper exposure equipment is a KrF stepper exposure equipment or an ArF stepper exposure equipment. The method according to claim 1,
Wherein said first etching is performed by reactive ion etching or inductively coupled plasma etching. ≪ RTI ID = 0.0 > 8. < / RTI > The method according to claim 1,
Wherein the insulating layer is a silicon oxide film or a silicon nitride film having a thickness of 100 to 9000 angstroms. The method according to claim 1,
Wherein the insulating layer is deposited using a plasma chemical vapor deposition process. ≪ RTI ID = 0.0 > 8. < / RTI > The method according to claim 1,
Wherein the nano patterns are any one of a circular array, a polygonal array having a square or more and a honeycomb array, and the array of the nano patterns is formed in a parallel array or a zigzag array. The method according to claim 1,
Wherein the nanopowder is formed with a diameter and an interval of 100 to 1000 nm. The method according to claim 1,
Wherein the second etching is performed by reactive ion etching or inductively coupled plasma etching. ≪ RTI ID = 0.0 > 8. < / RTI > The method according to claim 1,
And removing the second photosensitive layer after the second etching of the insulating layer. ≪ RTI ID = 0.0 > 21. < / RTI >
delete

Images