KR20090022698A - Method of coating silan layer on substrate - Google Patents

Abstract

A method for coating a silane layer on a substrate is provided to prevent misalignment between a substrate and other components in a manufacturing process of biochips. A method for coating a silane layer on a substrate comprises the following steps of: preparing the substrate(10); spin-coating the substrate with silane(300); and coupling a probe on a silane layer. The spin-coating includes the following steps of: coating a silane solution on the substrate; rotating the silane solution-coated substrate at a first speed and rotating again at a second speed; and solidifying the silane solution. The first speed is 500rpm, and the second speed is 2,000rpm.

Description

Method of coating silane layer on substrate {Method of coating silan layer on substrate}

The present invention relates to a method for manufacturing a biochip using spin coating.

Recently, with the development of the genome project, interest in biochips is increasing as the genome nucleotide sequences of various organisms are identified. Biochips are widely used for gene expression analysis, genotyping, detection of mutations and polymorphisms such as SNPs, protein and peptide analysis, potential drug screening, and drug development and manufacturing.

Such a biochip includes a plurality of probes for detecting a target sample, and the substrate was treated with a silane solution or the like to form the probes on the substrate more effectively. However, in the process of processing the silane solution on the substrate, unnecessary silane by-products may be formed, such as an alignment key, which may worsen the stability of the process in the manufacturing process of the biochip.

The problem to be solved by the present invention is to provide a process for producing a stable biochip.

The problem to be solved by the present invention is not limited to the above-mentioned problem, another task that is not mentioned will be clearly understood by those skilled in the art from the following description.

The manufacturing process of the biochip of the present invention for achieving the above technical problem includes providing a substrate, spin coating a silane layer on the substrate, and coupling a probe on the silane layer.

Other specific details of the invention are included in the detailed description and drawings.

According to the method of manufacturing a biochip according to embodiments of the present invention, a meniscus-type silane layer is not formed on the sidewall of the alignment key, so that a misalignment between the substrate and other equipments during the manufacturing process of the biochip is performed. alliegn) can be prevented. In addition, since the silane layer is not formed on the back surface of the substrate, malfunction in the substrate leveling in equipment such as a stepper may be prevented.

Advantages and features of the present invention and methods for achieving them will be apparent with reference to the embodiments described below in detail with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in various forms, and only the present embodiments are intended to complete the disclosure of the present invention, and the general knowledge in the art to which the present invention pertains. It is provided to fully convey the scope of the invention to those skilled in the art, and the present invention is defined only by the scope of the claims. Like reference numerals refer to like elements throughout.

When an element is referred to as being "connected to" or "coupled to" with another element, it may be directly connected to or coupled with another element or through another element in between. This includes all cases. On the other hand, when one device is referred to as "directly connected to" or "directly coupled to" with another device indicates that no other device is intervened. Like reference numerals refer to like elements throughout. “And / or” includes each and all combinations of one or more of the items mentioned.

Although the first, second, etc. are used to describe various elements, components and / or sections, these elements, components and / or sections are of course not limited by these terms. These terms are only used to distinguish one element, component or section from another element, component or section. Therefore, the first device, the first component, or the first section mentioned below may be a second device, a second component, or a second section within the technical spirit of the present invention.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In this specification, the singular also includes the plural unless specifically stated otherwise in the phrase. As used herein, “comprises” and / or “comprising” refers to the presence of one or more other components, steps, operations and / or elements. Or does not exclude additions.

1 is a view for explaining a protrusion formed on a substrate in the manufacturing process of the biochip according to the embodiments of the present invention.

Referring to FIG. 1, a plurality of protrusions 200 separated by a separation region 260 are formed on a substrate 10.

Substrate 10 may be a flexible or rigid substrate. Examples of flexible substrates to be applied include membranes or plastic films such as nylon and nitrocellulose. Examples of the rigid substrate include a silicon substrate and a transparent glass substrate made of soda lime glass. In the case of silicon substrates or transparent glass substrates, nonspecific binding hardly occurs during the hybridization process. In addition, the silicon substrate or the transparent glass substrate has an advantage that a manufacturing process and a photolithography process of various thin films that are already established and applied stably in a semiconductor device manufacturing process or an LCD panel manufacturing process and the like can be applied as they are.

The plurality of protrusions 200 separated by the separation regions 260 include an alignment key 250 and a probe cell active region 210. In this case, the probe cell active region 210 is a region capable of forming a probe cell by coupling with the probe (see FIG. 5).

A probe cell is defined as including a cell in which a plurality of probes that can hybridize with a target sample and detect a target sample are arranged. For example, the probe can be an antibody, enzyme, oligomeric probe, or the like.

An oligomer is a polymer composed of two or more monomers (covalently bonded), and may refer to a molecular weight of about 1,000 or less, but is not limited to the above numerical value. The oligomer may comprise about 2 to 500 monomers, preferably 5 to 30 monomers. The monomer may be a nucleoside, nucleotide, amino acid, peptide or the like depending on the type of probe immobilized on the probe array.

Nucleosides and nucleotides include known purine and pyrimidine bases, as well as methylated purines or pyrimidines, acylated purines or pyrimidines, and the like. In addition, nucleosides and nucleotides may include conventional ribose and deoxyribose sugars, as well as modified sugars in which one or more hydroxyl groups are substituted with halogen atoms or aliphatic groups, or functional groups such as ethers, amines, and the like. .

The amino acids may be L-, D-, and nonchiral amino acids of amino acids found in nature, as well as modified amino acids, amino acid analogs, and the like. A peptide refers to a compound produced by the amide bond between the carboxyl group of an amino acid and the amino group of another amino acid.

Thus, oligomeric probes may consist of two or more nucleosides, nucleotides, amino acids, peptides, and the like.

The alignment key 250 may be an alignment reference in a continuous biochip manufacturing process and a detection process using the biochip. For example, the substrate may be aligned in a mask, an exposure apparatus, or the like in photolithography, or in a dicing process in which the substrate is cut and separated into individual biochips. In addition, the cut individual chips may be used as an alignment reference in a detection process such as a scanning process after hybridization, as well as in the packaging of the cut individual chips.

The surface of the isolation region 260 may not include functional groups capable of coupling with the silane layer (see 300 in FIG. 2) or the probe (see 450 in FIG. 5) described below. In an embodiment of the present invention, the surface of the isolation region 260 may be the surface of the substrate 10. For example, in the case of a silicon substrate or a transparent glass substrate, the surface of the isolation region 260 may be a surface of the silicon substrate or the transparent glass substrate.

Although not shown in the drawing, in another embodiment of the present invention, a blocking film may be further formed between the protrusions 200. The blocking film does not include a functional group capable of coupling with the silane layer and the probe, and may be, for example, a fluorine silane film, a silicide film, a polysilicon film, or a Si, SiGe epitaxial film. In this case, the surface of the separation region may be the surface of the blocking film.

In one embodiment of the present invention, the protrusion 200 may be formed in a pattern of a film formed on the substrate 10.

The pattern of the film formed on the substrate 10 may be formed by, for example, forming a protrusion forming film on the substrate 10 and patterning the protrusion forming film.

For example, the film for forming the protrusion may be a silicon oxide film such as a Plasma Enhenced TetraEthylOrthoSilicate (PE-TEOS) film, a High Density Plasma (HDP) oxide film or a P-SiH4 oxide film, a thermal oxide film, a silicate such as hafnium silicate, zirconium silicate, silicon nitride film, Metal oxynitride films such as silicon oxynitride films, hafnium oxynitride films, zirconium oxynitride films, titanium oxide films, tantalum oxide films, aluminum oxide films, hafnium oxide films, zirconium oxide films, metal oxide films such as ITO, polyimides, polyamines, gold, silver, copper, and palladium Metal, such as these, or polymers, such as polystyrene, polyacrylic acid, and polyvinyl.

The protrusion forming film is a deposition method that is stably applied in a semiconductor manufacturing process or an LCD manufacturing process, for example, chemical vapor deposition (CVD), sub-atmospheric CVD (SACVD), low pressure CVD (LPCVD), and plasma enhanced CVD (PECVD). , Sputtering, or the like may be applied to the substrate 10.

Patterning the protrusion forming film may include forming a photoresist pattern on the protrusion forming film, etching the protrusion forming film using the photoresist pattern as an etching mask, and removing the photoresist pattern. Forming the photoresist pattern may proceed by forming a photoresist layer on the film for forming the protrusions, exposing and developing using a mask.

In addition, although not shown in the drawing, in another embodiment of the present invention, the protrusion may be formed of a (LOCal Oxidation of Silicon) oxide film in the substrate. For example, the LOCOS oxide film may be formed by forming an anti-fire pattern on the substrate and oxidizing the pattern exposed by the anti-oxidation pattern. The antioxidant pattern may be, for example, a nitride film or a stacked form of an oxide film and a nitride film.

2 is a view for explaining a silane layer spin-coated on a substrate in the manufacturing process of the biochip according to the embodiments of the present invention. 3A and 3B are diagrams illustrating a speed of rotating a substrate in a process of manufacturing a biochip according to embodiments of the present invention.

Referring to FIG. 2, the silane layer 300 is spin coated on the substrate 10. Forming the silane layer 300 on the substrate 10 may include applying a silane solution onto the substrate 10, rotating the substrate to which the silane solution is applied, and solidifying the silane solution.

First, a silane solution may be applied onto the substrate 10 on which the protrusion 200 is formed. The silane solution includes a functional group capable of coupling with the probe, and may be, for example, hygioxy silane, amino silane and the like. More specifically, the silane solution is N- (3- (triethoxysilyl) -propyl) -4-hydroxybutyramide (N- (3- (triethoxysilyl) -propyl) -4-hydroxybutyramide), N, N N, N-bis (hydroxyethyl) aminopropyl-triethoxysilane, acetoxypropyl-triethoxysilane, 3-glycidoxy propyltrimeth Methoxysilane (3-Glycidoxy propyltrimethoxysilane) and the like.

A functional group is defined as including a group that can be used as a starting point of an organic synthesis process. That is, groups such as probes such as synthetic oligomeric probes or monomers for in-situ synthesis, such as nucleosides, nucleotides, amino acids, peptides, etc. There is no specific limitation as long as it refers to a group capable of covalent or non-covalent coupling and can be coupled. Examples of the functional group include a hydroxyl group, an aldehyde group, a carboxyl group, an amino group, an amide group, a thiol group, a halo group, or a sulfonate group.

After the substrate 10 to which the silane solution is applied is rotated at a first speed, the substrate 10 may be subsequently rotated at a second speed. The substrate 10 may be rotated at a first speed to uniformly coat the silane solution applied on the substrate 10. In addition, by rotating the substrate 10 at a second speed, the excess silane solution may be removed so that the silane layer may be formed on the substrate 10 to a predetermined thickness, and further, the solvent may be removed. In order to effectively remove excess silane solution, the second speed may be faster than the first speed, and the first speed and the second speed may be, for example, 500 rpm and 2,000 rpm, respectively.

Referring to FIG. 3A, in the exemplary embodiment of the present invention, the silane solution may be coated by rotating the substrate 10 at a discontinuous change rate. For example, the substrate may be rotated at a first speed V1 for a predetermined time, and then the substrate may be subsequently rotated at a second speed V2 for a predetermined time. Here, in the discontinuously changing first speed V1 and the second speed V2, the second speed V2 may be faster than the first speed V1.

Referring to FIG. 3B, in another embodiment of the present invention, the silane solution may be coated by rotating the substrate at a continuously varying speed. For example, the silane solution may be coated by rotating the substrate at a linearly varying speed. Here, at a predetermined time, the second speed V2 may be faster than the first speed V1.

Solidification of the silane solution can be accomplished, for example, in a bake device. Depending on the size of the substrate 10 or the amount of the silane solution applied on the substrate 10, the bake time to the temperature may vary, for example, the substrate on which the silane layer 300 is applied for about 40 minutes at 120 ° C. You may bake (10).

Figure 4 is a schematic diagram for explaining the difference according to the coating method of the silane layer.

How the silane layer is formed according to the coating method will be described in more detail with reference to FIG. 4.

Referring to FIG. 4, when the silane layer 300 is formed by dip coating, the silane layer 300 is formed on the sidewall of the protrusion 200 due to the meniscus. It may be formed in a meniscus shape. In addition, since the dip coating includes dipping the substrate 10 in the silane solution, a region 300_b in which the silane solution is partially coupled may exist on the back surface 10_b of the substrate 10 on which the protrusion 200 is formed. have. Specifically, for example, when the substrate 10 is a silicon substrate, since a native oxide may exist on the back surface 10_b of the substrate 10, a region 330_b in which the silane solution is partially coupled may exist. .

On the other hand, in the case of spin coating, due to the rotation of the substrate 10, the silane layer 300 is not formed in the meniscus type on the sidewall of the protrusion 200. In addition, since the silane solution is applied only to the substrate 10 on which the protrusion 200 is formed, an unnecessary silane layer may not be formed on the back surface 10_b of the substrate 10. Furthermore, by coating the silane solution by rotation, it is possible to form a silane layer having a constant thickness.

Therefore, since the meniscus silane layer is not formed on the sidewall of the probe cell active region 210, the biochip may be manufactured so that unwanted probes are not formed on the substrate 10. In addition, since the meniscus silane layer or the like is not formed on the alignment key 250, the mask, the exposure apparatus, and the like may be accurately aligned in a continuous biochip manufacturing process such as, for example, a photography process. In addition, since the unnecessary silane layer 300_b is not formed on the back surface 10_b of the substrate 10, the substrate leveling may be performed without a malfunction, for example, in a stepper or the like.

5 is a view illustrating a biochip completed by coupling a probe in a process of manufacturing a biochip according to embodiments of the present disclosure.

Referring to FIG. 5, the probe 450 is coupled onto the probe cell active region 210. The probe cell active region 210 may be directly coupled to the probe 450 or may be coupled via the linker 410.

The linker 410 may include a first linker that facilitates coupling between the probe cell active region 210 and the probe 450 and a second linker that enables free interaction between the probe 450 and the target sample. Can be. Wherein if the length of the first linker is long enough to allow free interaction with the target sample, the second linker may not be interposed.

Coupling the linker 410 and the probe 450 may be coupling a probe such as a pre-synthesized oligomer probe, or may be coupling by in-situ synthesis of a probe monomer. .

6 is a view for explaining a silane layer formed on a substrate in the manufacturing process of the biochip according to another embodiment of the present invention.

Referring to FIG. 6, the silane layer 300 may be formed on the substrate 10 by the aforementioned spin coating. As described with reference to FIG. 4, in the manufacturing process of the biochip according to another embodiment of the present invention, unnecessary silane layer is not formed on the back surface of the substrate 10, for example, the substrate may be leveled without a malfunction in a stepper or the like, thereby providing a stable biochip. The manufacturing process can be performed.

7 is a view for explaining a biochip completed by coupling a probe in the manufacturing process of the biochip according to another embodiment of the present invention.

Referring to FIG. 7, the probe 450 is coupled to the partially activated silane layer region AR. The probe coupled to the activated silane layer region AR may deactivate the entire silane layer (see 300 in FIG. 6) and partially activate the deactivated silane layer 300a to activate the silane layer region AR. And the deactivated silane layer region NAR, and the probe 450 may be coupled to the activated silane layer region AR. In this case, the deactivation is defined as a meaning including a state in which the probes 450 to the linker 410 cannot couple to the silane layer (see 300 in FIG. 6).

The silane layer (see 300 in FIG. 6) can be deactivated by coupling a protecting group. A protecting group is defined as including a group that blocks the site to which it is coupled to participate in a chemical reaction, and deprotection means including a protecting group that is separated from the coupling site to allow the site to participate in a chemical reaction. Is defined. That is, since the protecting group can be acidlable or photolabile, the functional group can be deprotected by acid or light. For example, the photodegradable protecting group can be selected from a variety of positive photodegradable groups including nitro aromatic compounds such as o-nitrobenzyl derivatives or benzylsulfonyl, and specifically 6-nitroveratriryloxycarbonyl (NVOC), 2-nitro Benzyloxycarbonyl (NBOC), α, α-methyl-dimethoxybenzyloxycarbonyl (DDZ), and the like.

Light may be partially irradiated to the deactivated silane layer 300a through a mask to form an activated silane layer region AR and an inactivated silane layer region NAR. Specifically, light may be partially irradiated onto the deactivated silane layer 300a to deprotect the protecting group to form the activated silane layer region AR. In functional terms, the activated silane layer region AR is substantially the same as the probe cell active region to which the silane layer is applied in FIG. 2, and the inactivated silane layer region NAR is substantially the same as the isolation region in FIG. 2.

Coupling the probe 450 to the activated silane layer region AR may couple the activated silane layer region AR directly to the probe 450 and via the linker 41. You can also Coupling the linker 410 and the probe 450 may be coupling a probe such as a pre-synthesized oligomer probe, or may be coupling by in-situ synthesis of a probe monomer. .

 More detailed information about the present invention will be described through the following specific experimental examples and comparative examples, and details not described herein will be omitted because they are sufficiently technically inferred by those skilled in the art.

Experimental Example

A thermal oxide film was formed to a thickness of 1,000 Å on an 8-inch silicon substrate, and a photoresist film was formed to a thickness of 1.2 μm on the substrate. The photoresist film was exposed in a 365 nm wavelength projection exposure apparatus using a checkerboard type mask having a 1.0 μm pitch, followed by development with a 2.38% aqueous tetraMethylAmmonium Hydroxide solution to form a checkerboard horizontal and vertical A photoresist pattern was formed that exposes the intersecting straight regions. The thermal oxide film was etched using the photoresist pattern as an etching mask to pattern the protrusion including the probe cell array region and the alignment key.

10-20 ml of the silane solution was applied to the silicon substrate, and the suspension state was maintained for about 5 seconds while the silane solution was applied on the silicon substrate. After the substrate was rotated at 500 rpm for about 15 seconds, the substrate was continuously rotated at 15 rpm for 15 seconds. After the rotation was stopped and the substrate was stabilized for about 13 minutes at room temperature, the plate was transferred to a baking equipment and solidified at 120 ° C. for 40 minutes. The substrate was then washed with deionized water and then spin dried.

Comparative Example

The conditions different from the above experimental example were the same, but the silane solution was coated on the substrate on which the protrusions were formed by dip coating instead of the spin coating.

<Comparison of Experimental Example and Comparative Example>

8A and 8B are SEM top views of biochips before coupling of probes formed according to Comparative Examples and Experimental Examples, respectively.

Referring to FIGS. 8A and 8B, the alignment key in which the silane layer is formed is compared, and the alignment key A 'in the experimental example by spin coating is more than the alignment key A in the comparative example by dip coating. It can be seen that it is formed clearly. In addition, in the comparative example by the dip coating, it can be seen that there is a region B in which the silane solution is not evenly formed.

Although embodiments of the present invention have been described above with reference to the accompanying drawings, those skilled in the art to which the present invention pertains may implement the present invention in other specific forms without changing the technical spirit or essential features thereof. I can understand that. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive.

1 is a view for explaining a protrusion formed on a substrate in the manufacturing process of the biochip according to the embodiments of the present invention.

2 is a view for explaining a silane layer spin-coated on a substrate in the manufacturing process of the biochip according to the embodiments of the present invention.

3A and 3B are diagrams illustrating a speed of rotating a substrate in a process of manufacturing a biochip according to embodiments of the present invention.

Figure 4 is a schematic diagram for explaining the difference according to the coating method of the silane layer.

5 is a view illustrating a biochip completed by coupling a probe in a process of manufacturing a biochip according to embodiments of the present disclosure.

7 is a view for explaining a biochip completed by coupling a probe in the manufacturing process of the biochip according to another embodiment of the present invention.

8A and 8B are SEM top views of biochips before coupling of probes formed according to Comparative Examples and Experimental Examples, respectively.

(Explanation of symbols for the main parts of the drawing)

10: substrate 200: protrusion

210: probe cell active region

250: alignment key 260: separation area

300: silane layer 410: linker

450: probe

Claims (9)

Providing a substrate, Spin-coating a silane layer on the substrate, The method of manufacturing a biochip comprising coupling a probe on the silane layer. The method of claim 1, The spin coating may include applying a silane solution onto a substrate, rotating the substrate on which the silane solution is applied at a first speed, then rotating at a second speed, and solidifying the silane solution. Manufacturing method. The method of claim 2, Wherein said first speed is 500 rpm and said second speed is 2,000 rpm. The method of claim 3, wherein The method of manufacturing a biochip to rotate the substrate for 15 seconds each at the first speed and the second speed. The method of claim 2, Solidifying the silane solution comprises baking for 40 minutes at 120 ℃. The method of claim 1, And forming a plurality of protrusions separated by a separation region on the substrate, wherein the protrusions include an alignment key and a plurality of probe cell active regions. The method of claim 6, The surface of the separation region does not contain a functional group capable of bonding with the silane layer or the probe. The method of claim 6, Coupling the probe comprises coupling the probe onto the plurality of probe cell active regions. The method of claim 1, The silane layer is a method for producing a biochip comprising a hydroxyl group, an aldehyde group, a carboxyl group, an amino group, an amide group, a thiol group, a halo group or a sulfonate group.

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