KR101109082B1 - Reusable biochip and Using method of the same - Google Patents

Abstract

The present invention relates to a reusable biochip, wherein a reusable substrate having a measuring spot for measuring resistance between electrodes filled with a phase change material and a reaction spot having a binding material attached thereto are formed to be interchangeably attached to the reusable substrate. Replacement slide; characterized in that it comprises a.
In the present invention, in the biochip, it is possible to provide a slide of a replaceable type by using a reusable phase change material, increase the use efficiency of the biochip, and thereby reduce the manufacturing cost.

Description

Reusable biochip and using method of the same

The present invention relates to a structure of a reusable biochip and a method of using the biochip using the same.

A biochip is a chip in which biomolecules probes such as DNA and protein to be attached on a substrate are densely attached, and the gene expression patterns, gene defects, protein distribution, and reaction patterns in a sample can be analyzed. Biochips can be divided into microarray chips attached to a solid substrate and lab-on-a-chip attached to the microchannels according to the attachment form of the probe.

In such biochips, a system capable of detecting the binding of a target molecule to a probe immobilized on a substrate is required to determine whether a target molecule (target material) capable of binding a probe (binding material) is present in a sample. .

Currently, DNA chips for genetic analysis mostly label fluorescent dyes on sample DNA, react with probes on the chip, and detect fluorescent materials remaining on the chip surface using a confocal microscope or CCD camera. Use the method.

However, since such an optical detection method is difficult to miniaturize and the digitized output cannot be seen, much research is being conducted on the development of a new detection method capable of producing an electrical signal.

Accordingly, the present applicant simply and accurately detects bioinformation such as genes by measuring the resistance between electrodes connected to both ends of the phase change layer using a GST layer to detect the degree of crystallization of the phase change layer. A method for detecting a phase change biochip has been filed.

Referring to FIG. 1, the phase change biochip according to the above-described method may include a Si substrate, an electrode formed on the Si substrate, a phase change layer (GST) formed on the electrode, a protective film (SiO ₂ ) formed thereon, and the like. It consists of the organic material formed on the top and nanoparticles attached to the organic material.

In the fabrication of such phase change biochips, a lithography (LITHOGRAPHY) process is performed by using a MASK, and a GST / SiO ₂ layer is formed on the Ti / TiN unit electrode thus formed. The GST and SiO ₂ layer deposition equipment is a device that uses a magnetron sputtering method and is designed to deposit GST and SiO ₂ film.

The biochip designed as described above has the advantage of miniaturization and digitized output, but there is a problem that it cannot be reused once used even though the manufacturing process is complicated and expensive.

The present invention has been made to solve the above problems, by using a reusable substrate for the portion including the phase change layer and the electrode, and by using a replaceable slide for the portion where the bonding material and the target material combine, An object of the present invention is to provide a biochip and a method of using the same, which may increase the use efficiency of the biochip and thereby reduce manufacturing costs.

The present invention in order to solve the above problems is a reuse substrate for measuring the resistance between the electrode filled with a phase change material; And a replacement slide replaceably attached to the reusable substrate and having a bonding material attached thereto to provide a reusable biochip.

In addition, the reuse substrate includes a measuring spot for varying the resistance between the electrodes according to the degree of crystallization of the phase change material, the replacement slide, that the binding material includes a reaction spot for binding to the target material It features.

In addition, in the structure of the biochip according to the present invention, the reuse substrate and the replacement slide are preferably attached so that the measurement spot and the reaction spot coincide.

In addition, the reuse substrate according to the present invention may be configured to include a Si substrate, a Si Oxide layer and an electrode formed under the Si Oxide layer and a phase change layer that is stacked from the Si Oxide layer and embeds the electrode.

In addition, the sum of the thickness of the replacement slide and the glass substrate according to the present invention is preferably 630㎛.

The biochip according to the present invention can be used in the following manner.

Specifically, a reusable substrate includes a reusable substrate on which a measurement spot for measuring resistance between electrodes filled with a phase change material is formed, and a replacement slide which is replaceably attached to the reusable substrate, and on which a reaction spot on which a probe DNA is attached is formed. Detecting a biomaterial using a biochip, and attaching a binding material for inducing a target material to a reaction spot of the replacement slide; Attaching metal nanoparticles to the target material and inducing complementary binding to the binding material; Attaching the replacement slide to the reuse substrate; Irradiating a laser to induce crystallization of the phase change material; It can be configured by using a reusable biochip comprising a.

In the present invention, in the biochip, the portion including the phase change layer and the electrode uses a reusable substrate, and the portion in which the binding material and the target material are combined uses a slide that can be replaced, thereby increasing the use efficiency of the biochip. Through this, the manufacturing cost can be reduced.

1 is a conceptual diagram illustrating a conventional phase change biochip structure.
2 is a perspective view schematically showing the structure of a biochip according to the present invention.
3 is a schematic diagram illustrating laser light irradiation and electrical property detection for crystallization of a phase change layer of a phase change biochip according to the present invention.
4 is a plan view showing the electrode structure of the phase change biochip of the present invention.

Hereinafter, with reference to the accompanying drawings will be described in detail the configuration and operation according to the present invention. In the description with reference to the accompanying drawings, the same components are given the same reference numerals regardless of the reference numerals, and duplicate description thereof will be omitted. The terms first, second, etc. may be used to describe various components, but the components should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from another.

Referring to FIG. 2, the phase change biochip of the present invention may be interchangeably attached to the reuse substrate 100 and the reuse substrate on which a measurement spot S for measuring resistance between electrodes filled with a phase change material is formed. , It comprises a replacement slide 200 is formed with a reaction spot (P) attached to the binding material.

The reuse substrate 100 is sequentially formed from below the Si substrate 110, the Si oxide layer 120 formed on the Si substrate, the electrode 130 formed thereon, the phase change layer 140 formed thereon, formed thereon It is formed including the protective layer 150.

2 and 4, the electrode 130 serves to detect the degree of phase change of the phase change layer 140 by measuring the resistance between the electrodes in the phase change layer 140. The electrode pattern formed on the oxide layer 120 forms a measurement spot S at predetermined intervals so that the resistance between the electrodes is measured.

Depending on the degree of phase change in the phase change layer 140, the higher the crystalline fraction, the lower the resistance compared to the predetermined reference resistance. Will be shown.

The metal used as the electrode 130 is titanium (Ti), titanium nitride (TiN), tungsten (tungsten, W), gold (Au), aluminum (Aluminum, Al), tantalum (Tantalum, Ta), Tantalum nitride (TaN), molybdenum (Molybdenum, Mo), copper (Copper, Cu) may be exemplified, and any one having a conductive characteristic may be used without limitation.

The phase change layer 140 is a chalcogenide material composed of at least one element selected from the group consisting of Te, Se, Ge, Sb, Bi, Pb, Sn, As, In _, Ag, S, Si, P, and O. , Ge _a Sb _b Te _c (a, b, c are the atomic mole fractions, a + b + c = 1, 0 <a, b, c <1) or In _d Ag _e Sb _f Te _g (d, e, f and g each use an atomic mole fraction, d + e + f + g = 1, 0 <d, e, f, g <1), and it is particularly preferable to use Ge ₂ Sb ₂ Te ₅ . The phase change layer is a material that causes phase change due to crystallization when there is a change over a certain temperature in thermal change such as laser irradiation, and the phase change biochip of the present invention has the characteristics of such a phase change layer. It is to detect bio information such as genes.

The protective layer 150 is for preventing the phase change layer 140 from being damaged and is preferably formed of a SiO ₂ layer.

Replacement slide 200 is formed of the same material as the protective layer 150, the reaction spot (P) for attaching the bonding material is formed at a position corresponding to the side-spot (S). The reaction spot may be marked with a groove or a line so as to know where the binding material is attached.

In the preferred embodiment of the biochip according to the present invention, the sum of the thickness (t) of the protective layer 150 and the replacement slide 200 in the state in which the reuse substrate 100 and the replacement slide 200 are combined is 630 μm. It is preferred to be set to.

In particular, as shown in FIG. 3, when the focal point of the laser L of the DVD optical pickup apparatus is fixed to 0.6 mm, which is the thickness of the DVD, the laser focus of the DVD optical pickup apparatus is accurately changed. You can get to the floor. As a result, the energy of the laser light is sufficiently transmitted to the phase change layer.

In connection with the manufacture of the biochip according to the present invention as described above, the process of forming the reuse substrate 100 is as follows.

(1) Ti / TiN electrode 130 forming process

The Si oxide layer 120 is formed by exposing the Si substrate 110 to the atmosphere, and Ti / TiN is deposited in-situ thereon. Next, a lithography (LITHOGRAPHY) process is performed using the MASK. Here, an ashing process is performed using N ₂ / Ar gas in an ICP method to improve the degree of removal of the photoresist. As a result, a Ti / TiN unit electrode is formed on the Si oxide layer 120.

(2) Phase Change Layer 140 Deposition Process

The phase change layer 140 is formed on the Si oxide layer 120 on which the electrode 130 is formed. The phase change layer 140 is deposited to a thickness that completely fills the electrode 140, starting from the Si oxide layer 120. The deposition equipment of the phase change layer 140 is designed to deposit a phase change layer by using a magnetron sputtering method. Here, the phase change layer 140 to be deposited is Ge ₂ Sb ₂ Te _{5 which} is a chalcogenide compound, and the process conditions are Base Pressure <5.0X10 ^-6 Torr, Power = DC 10 W, and the like.

(3) Process of forming protective layer 150

After the phase change layer 140 is deposited, a protective layer is formed. The protective layer is formed of a transparent material through which a laser can be transmitted, and is preferably formed of SiO ₂ .

Next, an example of a manufacturing process of the replacement slide 200 attached to the recyclable substrate will be described.

(1) Glutaraldehyde (glutaradehyde) (160) after the process of attaching the binding material 170

A SiO ₂ substrate of the same material as the protective layer 150 of the reused substrate 100 is immersed in a glutaraldehyde solution (2%) and placed in a sealed container and stored at room temperature for 2 to 4 hours. After this step, wash with DI water and dry with nitrogen gas. This completes the surface treatment for attaching the bonding material on the replacement slide 200.

The replacement slide 200 of which the surface treatment is completed is washed with 0.2% SDS for 10 minutes. Subsequently, the preliminary phase change biochip is washed twice with tertiary distilled water for 5 minutes. Next, the reaction was stirred for 10 minutes in a solution of 0.7 g sodium borohydride and 500 ml reduction solution. The mixed solution should be prepared just before use. Then wash for 5 minutes with tertiary distilled water. Then wash for 5 minutes with 0.2% SDS. Then, washed three times with 1 minute distilled water. Then dry using N ₂ gas or centrifuge. Subsequently, the replacement slide 200, which has been dried, is placed in a slide box containing a moisture absorbent and stored in a desiccator. This attaches the bonding material 170 to the replacement slide.

(2) Process of Attaching Target Material 180 Marked with Metal Nanoparticles 190

A process for attaching the target material 180 represented by the metal nanoparticles 190 to the bonding material attached to the replacement slide 200 described above will be described below. Metal nanoparticles, such as gold, silver, copper may be used.

First, 5 μl of Mycobacteria Genotyping PCR Product, streptavidin conjugated AuNP (40 nm diameter) and 80 μl of hybridization buffer were added to 500 microtubes. Create a Hybridization Mixture. The mixture is then mixed using a vortex and centrifuged to remove the solution from the lid. Subsequently, the replacement slide 200 is placed in a hybridization chamber filled with water at the bottom. The hybridization mixture is then injected with care to avoid bubbles through the sample inlets present in the cover covering the replacement slide. Then close the cover of the hybridization chamber. Subsequently, the mixture is placed in a hybridizer at room temperature and reacted for 1 hour. Then remove the cover of the replacement slide after hybridization. The washing is then performed by shaking or stirring at 50 rpm for 2 minutes in a washing buffer. Next, washing is performed by shaking or stirring at 50 rpm for 1 minute in distilled water. The replacement slide is then centrifuged at 1000 rpm for 1 minute or dried using compressed air. As a result, the target material 180 represented by the metal nanoparticles 190 is attached to the binding material 170.

Hereinafter, an application example of a method of using a biochip manufactured by the above method will be described.

The reuse substrate 100 is inserted into a biochip detection device (not shown), and the replacement slide 200 to which the target material is coupled is attached. Attachment of the reuse substrate 100 and the replacement slide 200 is preferably to always be in place by installing a jig or the like in the position where the reuse substrate 100 is inserted.

Subsequently, when the laser for inducing the crystallization of the phase change layer 140 is irradiated using the DVD optical pickup device, the degree of phase change varies according to the presence or absence of metal nanoparticles. When the target material having a base sequence complementary to the binding material is attached at a high density, the degree of phase change of the phase change layer will be reduced since the laser is blocked by the metal nanoparticles of the target material. On the other hand, when the target material having a base sequence complementary to the binding material is attached at low density, the degree of phase change of the phase change layer is small because the laser is blocked by the metal nanoparticles of the target material. Will make a lot. Thereby, the biochip to which the laser was irradiated is completed.

The bio-chip irradiated with the laser may apply a voltage to measure resistance between the electrodes. Depending on the degree of phase change of the phase change layer, the higher the crystalline fraction, the lower the resistance than the predetermined reference resistance. The reference resistance can be determined by experiment.

After measuring the electrical properties to detect the presence or absence of the target material, the process for reusing the biochip is as follows.

The measured biochip removes the replacement slide 200 and irradiates a laser for returning the phase change layer 140 to its original state (amorphous) to the reuse substrate 100. The laser for reuse is irradiated with a high output (30mW ~ 70mW) and a low pulse compared to the output of the laser for the phase change layer 140 crystallization.

When the phase change layer 140 returns to the original amorphous state by laser irradiation, a process of attaching a binding material that can complementarily bind to the target material to be detected is performed on the replacement slide 200. Complementary bonding of the target material to the replacement slide is attached to the reused substrate and after the above-described process, the electrical properties between the electrodes are detected.

In the detailed description of the present invention as described above, specific embodiments have been described. However, various modifications are possible within the scope of the present invention. The technical idea of the present invention should not be limited to the embodiments of the present invention but should be determined by the equivalents of the claims and the claims.

100: reused substrate 110: Si substrate
120: Si oxide layer 130: electrode
140: phase change layer 150: protective layer
160: glutaraldehyde 170: binding material
180: target material 190: metal nanoparticles
200: replacement slide P: reaction spot
S: measuring spot

Claims (8)

A reuse substrate for measuring resistance between electrodes filled with a phase change material; And
And a replacement slide replaceably attached to the reusable substrate and to which a bonding material is attached.
The method of claim 1, wherein the reuse substrate,
Reusable biochip comprising a measurement spot for varying the resistance between the electrodes according to the degree of crystallization of the phase change material.
The method of claim 2, wherein the replacement slide,
Reusable biochip, characterized in that the binding material comprises a reaction spot for binding to the target material.
The method of claim 3, wherein the reuse substrate and the replacement slide,
Recyclable biochip, characterized in that the measuring spot and the reaction spot is attached to match.
The method according to any one of claims 1 to 4, wherein the reuse substrate,
Si substrate;
Si Oxide layer;
An electrode formed on the Si Oxide layer;
A phase change layer stacked from the Si oxide layer and filling the electrode; And
Reusable biochip comprising a protective layer formed on the phase change layer.
6. The method of claim 5,
And the sum of the protective layer and the replacement slide thickness is 630 μm.
Bio using a reusable biochip comprising a reusable substrate having a measuring spot for measuring the resistance between the electrode filled with a phase change material and a replacement slide that is replaceably attached to the reusable substrate and a reaction spot having a binder attached thereto is formed. Detect information,
Attaching a binding material for inducing a target material to the reaction spot of the replacement slide;
Attaching metal nanoparticles to the target material and inducing complementary binding to the binding material;
Attaching the replacement slide to the reuse substrate;
Irradiating a laser to induce crystallization of the phase change material; And
Measuring the resistance between the electrodes; method comprising using a reusable biochip.
The method of claim 7, wherein after measuring the resistance between the electrodes,
Removing the replacement slide; And
Irradiating a laser on the reuse substrate to return the phase change material to its original state.

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