KR102013996B1 - Product method of lab on a chip - Google Patents

Abstract

The present specification discloses a microfluidic analysis chip and a method of manufacturing the same, which are not limited by the type of protein and the method of bonding the chip. Microfluidic analysis chip manufacturing method according to the present specification comprises the steps of (a) manufacturing a chip housing in which the microchannels for the main channel and a plurality of subchannels are formed; (b) performing a surface treatment for fixing a reactant on the surface of the microchannel for the main channel; And (c) injecting a reactant through the main channel microtubules or the subchannel microtubules.

Description

Microfluidic analysis chip manufacturing method {PRODUCT METHOD OF LAB ON A CHIP}

The present invention relates to a method for manufacturing a microfluidic analysis chip, and more particularly, to a method for manufacturing a microfluidic analysis chip having improved function and accuracy compared to a conventional microfluidic analysis chip.

Biochip refers to the accumulation of biomolecules such as DNA and protein on a small substrate made of glass, silicon, or nylon.In this case, when DNA is accumulated, it is called DNA chip. This is called. Biochips can also be broadly divided into microarray chips and microfluidics chips.

Microarray chips are biochips that can arrange thousands or tens of thousands or more of DNA or proteins at regular intervals, process analytes, and analyze their binding patterns. Microfluidics chip is a biochip that analyzes the reaction of various kinds of biomolecules or sensors integrated in the chip while flowing a small amount of analyte. It is also called a lab on a chip. It is a cutting-edge technology that combines the functions of sensors, pumps, valves, reactors, extractors, separation systems, etc., which are essential for the sample preparation process of automated analytical devices used for the analysis of biochemicals.

Looking more closely at Lab-on-a-Chip, Lab-on-A-Chip is designed to perform sample injection, pretreatment, chemical reactions, separation / analysis, etc., which are performed at the laboratory level in order to analyze chemicals and biochemicals. One microanalyzer.

Lab-on-a-chip technology combines micro flow control technology with MEMS micromachining technology to accurately transfer, distribute and mix samples from several picoliters (pl) to tens of microliters (μl). It is a core technology.

Lab-on-a-Chip, which uses trace amounts of samples and analyzes chemical components quickly and easily, is widely used to select useful new drugs out of many new drug candidates at high speed.In recent years, lab-on-a-chip is used to detect environmental pollutants and diagnose diseases. Kind of lab-on-a-chip is under research and development.

Unlike micro-array chips such as DNA chips and protein chips, lab-on-a-chips are still at the stage of R & D in the world, and commercialization is limited and small-scale. In the case of the lab-on-a-chip, the network of microchannels is simple and the reaction process is implemented at a stage not complicated.

Republic of Korea Patent Publication No. 10-2010-0060723 (2010.06.07)

The present specification is to provide a microfluidic analysis chip and a method for manufacturing the same, which are not limited by the type of protein and the method of bonding the chip.

The present specification is not limited to the above-mentioned task, and other tasks not mentioned will be clearly understood by those skilled in the art from the following description.

Microfluidic analysis chip manufacturing method according to the present specification for solving the above problems, (a) manufacturing a chip housing formed with a microtube for the main channel and a plurality of subchannels; (b) performing a surface treatment for fixing a reactant on the surface of the microchannel for the main channel; And (c) injecting a reactant through the main channel microtubules or the subchannel microtubules.

According to an embodiment of the present disclosure, the step (a) includes the steps of manufacturing a lower chip and a lower chip chip plate formed with a micro-tube for the main channel and a plurality of sub-channels; And combining the chip lower plate and the chip upper plate.

In this case, in the bonding step, the lower chip plate and the upper chip plate may be combined using at least one method of heat treatment, ultraviolet treatment, and chemical treatment.

According to an embodiment of the present disclosure, the step (a) may include a cap or a valve blocking the main tube microtube or the subchannel microtube and the outside of the main tube microtubule or the subchannel microtubule. Forming; may further include.

According to an embodiment of the present disclosure, the step (b) may include the first and second subchannel microtubules of the second subchannel of the plurality of subchannel microtubules and the plurality of subchannel microtubules. Closing the ends of the remaining microchannels except for the subchannels with a cap or a valve; And (i) a first point at which the first subchannel microtube and the main channel microtube are connected, and (ii) the second subchannel microtube and the main channel. Injecting a surface treatment solution through the first subchannel microtube or the second subchannel microtube for the surface treatment for fixing the reactant to the surface of the region between the second points to which the microtubules are connected; It may include.

According to the exemplary embodiment of the present specification, the step (c) may include ends connected to both ends of the main tube microtubules and to the outside of the remaining microchannel tubules except for the first and second subchannel microtubules. Closing with a cap or valve; And (i) a first point at which the first subchannel microtube and the main channel microtube are connected, and (ii) the second subchannel microtube and the main channel. And injecting a reactant solution through the first subchannel microtube or the second subchannel microtube to fix the reactant on the surface of the region between the second points to which the dragon microtube is connected. .

The method for manufacturing a microfluidic analysis chip according to the present specification includes (d) (i) a cap or valve of the microtube for the first or second subchannel and (ii) a main channel adjacent to the cap or valve selected from (i). Opening the cap or valve of the dragon microtubule; And (e) injecting a removal solution for removing a non-fixed reactant on the surface of the region between the first point and the second point through the subchannel microtubule or the mainchannel microtubule opened in (d). It may further include.

The method for using a microfluidic analysis chip according to the present specification includes: (a) a main tube microtube for providing a space for reacting with a reagent while a sample injected from a sample inlet formed at one end moves to the other end, and one end of the main The microfiber is connected to a side surface of the channel microtubule, and the other end includes a plurality of subchannel microtubes connected to the outside of the chip housing, and a chip housing surrounding the main channel microtube and the plurality of subchannel microtubes. A fluid analysis chip comprising: (i) a first point of a plurality of subchannel microtubes in which a first subchannel microtube and the main channel microtube are connected; The fine substance in which a reactant or a hydrogel is fixed on the surface of the region between the second subchannel microtube for the second subchannel and the main channel microtube among the plurality of subchannel microtubes Injecting a sample through said sample injection port of the analysis chip body; (b) (i) opening the cap or valve connected to both ends of the microchannel for the main channel, and (ii) closing the ends connected to the outside of the plurality of subchannel microtubes with a cap or valve; And (c) injecting a washing solution for washing the sample remaining in the microchannel for the main channel without reacting with the reactant through one of both ends of the main tube for the microchannel.

Microfluidic analysis chip manufacturing method according to the present specification for solving the above problems, (a) manufacturing a chip housing formed with a microtube for the main channel and a plurality of subchannels; (b) closing both ends of the main tube microtubules and the ends connected to the outside of the other subchannel microtubules except the first and second subchannel microtubules with a cap or a valve; And (c) injecting a hydrogel through the first or second subchannel microtubules. Further, (b-1) may be a surface treatment for fixing the hydrogel on the surface of the microchannel for the main channel; may further include.

The microfluidic analysis chip fabrication method according to the present disclosure is based on (d) at least one of measured impedance, magnetic field, and optical value of the target region, whether (i) the sample or the reagent is reached, (ii) the sample Or (iii) connecting a control unit for determining at least one of the flow rate of the reagent, (iii) the amount of the sample or the reagent, and (iv) the sample or the type of the reagent to the main channel microtubule. It may further include.

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

According to one aspect of the present specification, a desired local region of the surface of the microchannel for the main channel may be fixed with a reactant or a hydrogel. This allows more precise reaction of reagents and samples in the desired area.

According to another aspect of the present specification, since the reaction material such as protein is fixed to the main channel microtubules after the upper chip plate and the lower chip plate are combined in the manufacturing process, possibility of denaturation of the reactant such as protein by heat treatment or the like during the manufacturing process This becomes very low.

According to another aspect of the present disclosure, it is possible to detect a variety of materials through the surface treatment of the local region of the microtubules for the main channel with different materials. In addition, through the hydrophilic and hydrophobic material it is possible to control the flow rate or to limit the flow of the microfluid for each section.

Effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the following description.

1 is a flowchart illustrating a method of manufacturing a microfluidic analysis chip according to an exemplary embodiment of the present specification.
2 is a plan view and a cross-sectional view of a microfluidic analysis chip manufactured according to the microfluidic analysis chip manufacturing method of the present specification.
3 is an exemplary view in which a cap is formed on the microfluidic analysis chip according to the present specification.
4 is an exemplary view of injecting a surface treatment solution and a reactant solution according to one embodiment of the present specification.
5 is an exemplary view of injecting a surface treatment solution and a reactant solution according to another embodiment of the present specification.
6 is an exemplary view of injecting a surface treatment solution and a reactant solution according to another embodiment of the present specification.
7 is an enlarged cross-sectional view of a part of the microchannel for the main channel according to the present specification.
8 is an exemplary view of a method for removing unnecessary reactants according to the present specification.
9 is an exemplary view for washing the inside of the microtubules for the main channel according to an embodiment of the present disclosure.
10 is an enlarged view of a portion of a microfluidic analysis chip having a plurality of electrodes according to the present specification.
11 and 12 are exemplary views of a microchannel for a subchannel including a valve according to an embodiment of the present disclosure.

Advantages and features of the invention disclosed herein, and methods of achieving them will be apparent with reference to the embodiments described below in detail in conjunction with the accompanying drawings. However, the present disclosure is not limited to the embodiments disclosed below, but may be implemented in various forms, and the present embodiments are merely provided to make the disclosure of the present disclosure complete, and those of ordinary skill in the art to which the present disclosure belongs. It is provided to fully inform the skilled person of the scope of the present specification, and the scope of the present specification is defined only by the scope of the claims.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the scope of the present disclosure. In this specification, the singular also includes the plural unless specifically stated otherwise in the phrase. As used herein, "comprises" and / or "comprising" does not exclude the presence or addition of one or more other components in addition to the mentioned components. Like reference numerals refer to like elements throughout, and "and / or" includes each and all combinations of one or more of the mentioned components. Although "first", "second", etc. are used to describe various components, these components are of course not limited by these terms. These terms are only used to distinguish one component from another. Therefore, of course, the first component mentioned below may be a second component within the technical spirit of the present invention.

Unless otherwise defined, all terms used in this specification (including technical and scientific terms) may be used in a sense that can be commonly understood by those skilled in the art to which the present specification belongs (hereinafter, referred to as a person skilled in the art). . In addition, terms that are defined in a commonly used dictionary are not ideally or excessively interpreted unless they are specifically defined clearly.

The spatially relative terms " below ", " beneath ", " lower ", " above ", " upper " It can be used to easily describe a component and its correlation with other components. Spatially relative terms are to be understood as including terms in different directions of components in use or operation in addition to the directions shown in the figures. For example, when flipping a component shown in the drawing, a component described as "below" or "beneath" of another component may be placed "above" the other component. Can be. Thus, the exemplary term "below" can encompass both an orientation of above and below. Components may be oriented in other directions as well, so spatially relative terms may be interpreted according to orientation.

Hereinafter, with reference to the accompanying drawings will be described an embodiment of the present invention;

1 is a flowchart illustrating a method of manufacturing a microfluidic analysis chip according to an exemplary embodiment of the present specification.

Referring to FIG. 1, in the method of manufacturing a microfluidic analysis chip according to an embodiment of the present specification, a step of manufacturing a chip housing in which a microtube for a main channel and a microtube for a plurality of subchannels is formed (S10), for the main channel Surface treatment for fixing the reaction material on the surface of the microtubules (S20) and the step of injecting the reactants through the microchannels for the main channel or subchannels (S30).

2 is a plan view and a cross-sectional view of a microfluidic analysis chip manufactured according to the microfluidic analysis chip manufacturing method of the present specification.

Referring to FIG. 2, the microfluidic analysis chip 100 according to the present disclosure may include a chip housing 110, a main tube microtube 120, and a plurality of subchannel microtubes 130.

The chip housing 110 may be made of a polymer material such as plastic.

In the present specification, the microchannel 120 for the main channel is a space into which a sample such as blood and urine is introduced and moved, and a reaction chamber for reacting with a reagent in the main channel microtubule. This can be formed. In addition, both ends of the main channel microtubule 120 may be connected to the outside of the chip housing 110. The exterior does not necessarily mean a space that is physically spaced relative to the end of the housing. Since the main tube microtubules 120 are to be sampled, one end should be connected to the outside for sample input. In addition, after the sample injected into the main tube 120 for the main channel reacts with the reagent, the sample should be moved to the opposite side to check the result, and the result should be confirmed from the outside. In the drawings, an inlet and an outlet of the main tube microtube 120 are illustrated on the top surface of the chip housing, but the microfluidic analysis chip according to the present specification is not limited to the drawings. It is apparent that the inlet and the outlet may be formed in various ways such as the top, bottom, side of the chip housing. Therefore, in the present specification, both ends of the main channel microtubule 120 are connected to the outside of the chip housing 110, and the reaction of the sample and the reagent made in the main channel microtubule 120 is confirmed from the outside. It should be understood in various forms so that it can be made.

One end of the plurality of subchannel microtubes 130 may be connected to a side surface of the main channel microtube 120, and the other end thereof may be connected to the outside of the chip housing 110. In the present specification, the 'side surface of the microchannel for the main channel' means a side surface based on the moving direction of the fluid flowing in the microchannel for the main channel. Therefore, the microchannels for the subchannels do not need to be vertically connected to the surface of the microchannels for the main channel, and mean various forms in which the inside of the main channel microtubules and the inside of the subchannel microtubules can be connected. do. On the other hand, Figure 2 is shown an embodiment having two micro-channels 130 for the sub-channel, but is not limited to the embodiment shown in the figure and the number varies as necessary.

 The chip housing 110 may be manufactured integrally by an injection method, or may be manufactured in a method in which the chip lower plate 111 and the chip upper plate 112 are combined. When the chip housing 110 is divided into a chip lower plate 111 and a chip upper plate 112, the step S10 may include a main channel microtube 120 and a plurality of subchannel microtubes 130. It may include the step of manufacturing the chip lower plate 111 and the chip upper plate 112 and the step of coupling the chip lower plate 111 and the chip upper plate 112. In the combining step, the lower chip plate 111 and the upper chip plate 112 may be combined using at least one of heat treatment, ultraviolet light treatment, and chemical treatment. In this case, the microchannel 120 for the main channel may be formed on a surface where the chip lower plate 111 and the chip upper plate 112 are coupled to each other.

3 is an exemplary view in which a cap is formed on the microfluidic analysis chip according to the present specification.

Referring to FIG. 3, it can be seen that the cap 140 is formed at the ends of the main channel microtubule and the subchannel microtubule. The cap 140 is configured to open or close the main channel microtubule or the subchannel microtubule, and when the cap is closed, the microtubule is blocked from the outside so that air does not flow or fluid enters. . In this case, the step S10 may further include forming a cap or a valve blocking the main channel microtube or the subchannel microtube and the outside in the main channel microtubule or the subchannel microtubule. have. On the other hand, how to use the cap 140 will be described below.

 According to one embodiment of the present specification, (i) a first point of a first sub-channel microtube and the main channel microtubule among the plurality of subchannel microtubes in the entire region of the main channel microtubule And (ii) the protein-fixing material is chemically treated on the surface of the region between the second point of the plurality of subchannel microtubes connected to the second subchannel microtube and the main channel microtube. Can be.

According to an embodiment of the present disclosure, the step S20 may include remaining ends of the main tube microtubes, except for the first subchannel microtube and the second subchannel microtube among the plurality of subchannel microtubes. Closing an end connected to the outside of the microchannel for the subchannel with a cap or a valve, and (i) connecting the first subchannel microtube and the main channel microtubule to an entire region of the main channel microtubule; (Ii) the first subchannel microtube for surface treatment for fixing a reactant on the surface of the region between the first point and (ii) the second point where the second subchannel microtube and the main channel microtube are connected; Or injecting the surface treatment solution through the second microchannel for the subchannel.

According to an embodiment of the present disclosure, the step S30 may include caps or ends connected to both ends of the main tube microtubules and to the outside of the remaining microchannel tubules except for the first and second subchannel microtubules. (I) a first point to which the first subchannel microtube and the main channel microtubule are connected; and (ii) the second subchannel micrometer. Injecting a reactant solution through the first subchannel microtubule or the second subchannel microtubule to fix the reactant on the surface of the region between the tubule and the second point to which the main tubular microtube is connected; It may include.

4 is an exemplary view of injecting a surface treatment solution and a reactant solution according to one embodiment of the present specification.

Referring to FIG. 4D for a while, it may be confirmed that the surface treatment and the reactant are fixed to a part of the microchannel for the main channel. In the present specification, for convenience of understanding, (i) a point where the first subchannel microtube and the main channel microtube are connected to each other in the entire region of the main channel microtubule, One point ', and (ii) a point where the second subchannel microtube and the main channel microtube are connected to the second point among the plurality of subchannel microtubes. The first subchannel microtubule is a subchannel microtubule corresponding to the first point, and the second subchannel microtubule is a subchannel microtubule corresponding to the second point.

First, the cap formed in the microchannel 120 for the main channel is closed in FIG. 4A, and the cap formed in the microchannel 130 for the subchannel is opened. Caps cover both ends of the main tube microtubes and the ends connected to the outside of the microchannels for the other subchannels except the first and second subchannel microtubes. In FIG. 4, since only the microtubes for the first and second subchannels are shown, the state of blocking the remaining microchannels for the subchannels is not illustrated. However, according to various embodiments, other microchannels for the subchannels may be formed in addition to the microtubes for the first and second subchannels. In this case, when the surface treatment solution for fixing the reactant is injected through the sub-channel microtube 130, the surface between the first point and the second point is treated as shown in FIG. The surface treatment solution may be BSA (bovine serum albumin), HEC (hydroxyethyl-cellulose), MC (methyl-cellulose), PVA (poly (vinyl alcohol), PP (pluronic polyol) or DS (dextransulfate). As shown in (c) of FIG. 4, the reactant solution may be injected through any one of an end connected to the outside of the first microchannel for the subchannel and an end connected to the outside of the second microchannel for the second subchannel. As a result, the reactant is fixed to the surface between the first point and the second point as shown in (d) of FIG.

The reactant may be a substance that chemically reacts with a specific substance or an antigen-antibody reactant, or a protein that binds to a specific component. That is, it may be a variety of materials that react with the target material according to the properties of the material to be found in the sample.

5 is an exemplary view of injecting a surface treatment solution and a reactant solution according to another embodiment of the present specification.

(A) and (b) of FIG. 5 are the same as (a) and (b) of FIG. Therefore, the description of the repeated parts will be omitted and the difference will be described from (c) of FIG. 5. Referring to FIG. 5C, the cap formed in the microchannel 120 for the main channel is opened, and the cap formed in the microchannel 130 for the subchannel is closed. The reactant solution may be injected through any one of both ends of the main tube microtubule 120. In this case, since the reactant fixing material is surface treated only between the first point and the second point, the reactant is fixed to the surface between the first point and the second point as shown in FIG.

6 is an exemplary view of injecting a surface treatment solution and a reactant solution according to another embodiment of the present specification.

First, the cap formed in the microchannel 120 for the main channel is opened in FIG. 6A, and the cap formed in the microchannel 130 for the subchannel is closed. When the surface treatment solution for fixing the reactant is injected through the main channel microtubule 120, the surfaces of all regions of the main channel microtubule are surfaced with the reactant fixing substance as shown in FIG. Processing. Next, referring to FIG. 6C, the cap formed in the main channel microtubule 120 is closed, and the cap formed in the subchannel microtubule 130 is opened. The reactant solution may be injected through any one of an end connected to the outside of the first microchannel for the subchannel and an end connected to the outside of the second microchannel for the second subchannel. As a result, the reactant is fixed to the surface between the first point and the second point as shown in FIG.

4 to 6 illustrate embodiments in which the surface of the local region is fixed with a reactant. 4 to 6 have been described with reference to an embodiment in which two subchannel microtubules are formed for simplicity and convenience of description, but additional microchannels for subchannels other than the first and second subchannel microtubules are configured. In this case, a plurality of reactant fixing regions can be made even in one main channel microtubule. In this case, the use of heterogeneous reactants enables analysis of multiple target substances within one channel.

On the other hand, the reactant may be fixed beyond the required first point or second point, and may remain on the surface of the microchannel for the subchannel.

7 is an enlarged cross-sectional view of a part of the microchannel for the main channel according to the present specification.

In the manufacturing method according to the present disclosure, the surface treatment solution and the reactant solution are injected through the microchannel for the subchannel corresponding to the desired local region by blocking the main channel microtubule with a cap or a valve, but as shown in FIG. 7. The surface treatment solution or the reactant solution may deviate from the expected first point or second point. In FIG. 7, the left side of the first subchannel microtubule is expressed as an area where the reactant is undesirably fixed. In addition, it was expressed that some of the reactants remained on the surface of the microtubules for the first subchannel. The fabrication method according to the present disclosure enables the removal of reactants in the unwanted region.

8 is an exemplary view of a method for removing unnecessary reactants according to the present specification.

Referring to FIG. 8A, the protein is fixed between the first point and the second point according to the method of FIG. 6. At this time, it is assumed that the unwanted reactant is to be removed from the surface of the microtubules for the first subchannel, the surface of the microtubules for the second subchannel, the left side of the first point and the right side of the second point. To this end, as shown in (b) of FIG. 8, the left end of the main tube microtubule and the cap of the first subchannel microtubule are opened, and the right end of the main channel microtubule and the second subchannel microtubule The cap is closed. Then, the removal liquid is injected through the left end of the main channel microtubule or the first subchannel microtubule. This removes unnecessary reactants present on the surface of the first microchannel for the subchannel and to the left of the first point. Next, as shown in FIG. 8C, the right end of the main channel microtubule and the cap of the second subchannel microtubule are opened, and the left end of the main channel microtubule and the first subchannel microtubule. Cap should be closed. Then, the removal liquid is injected through the right end of the main channel microtubule or the second subchannel microtubule. This removes unnecessary reactants present on the surface of the microtubules for the second subchannel and to the right of the second point. As a result, as shown in (d) of FIG. 8, unnecessary reactants may be removed through the removal liquid.

Meanwhile, FIG. 8 illustrates an embodiment in which two microchannels for subchannels are provided. However, when more than two subchannel microtubules are provided, another subchannel microtubule may serve as the main channel microtubules. For example, four subchannel microtubules are provided, and the points of the main channel microtubules corresponding to each of the subchannel microtubules are referred to as a first point, a second point, a third point, and a fourth point. I'll name it. In this case, it is assumed that the reactant is fixed between the second point and the third point, and to remove unnecessary reactants in the remaining part. To this end, the microtubules for the first subchannel and the microtubules for the second subchannel are opened, and the rest are closed, and a removal liquid is injected through the microtubules for the first subchannel or the microtubules for the second subchannel. Then, the third subchannel microtubule and the fourth subchannel microtubule are opened, and the rest are all closed, and the removal liquid is injected through the third subchannel microtubule or the fourth subchannel microtubule. This will remove unnecessary reactants in the remaining regions except for the reactants fixed between the second and third points.

That is, (i) the cap or valve of the microtube for the first or second subchannel and (ii) the cap or valve of the microchannel for the main channel adjacent to the cap or valve selected from (i) or the cap for the subchannel or Opening the valve. In addition, the removing liquid is injected to remove the reactant which is not fixed to the surface of the region between the first point and the second point through the open subchannel microtube or the main channel microtube.

The removal solution selects a suitable solution according to the binding method of the material and the surface to be removed, for example, the protein removal solution is 15 g glycine, 1 g SDS, 10 ml Tween20, Adjust pH to 2.2, Bring volume up to 1 L With ultrapure water solution or 20 ml SDS 10%, 12.5 ml Tris HCl pH 6.8 0.5M, 67.5 ml ultra pure water, 0.8 ml ß-mercaptoethanol solution can be used.

On the other hand, the sample is reacted (or combined) with the reactant, but the sample that exceeds the amount of the unreacted substance or reactant may remain in the microchannel for the main channel. Therefore, there is a need to wash the microchannel for the main channel.

9 is an exemplary view for washing the inside of the microtubules for the main channel according to an embodiment of the present disclosure.

Referring to FIG. 9A, first, a sample is injected. The sample was expressed as containing a substance that binds to the reactants. Next, referring to FIG. 9 (b), it can be seen that all of the samples reacted with the reactants, but some of the samples remained in the microchannels for the main channel. Next, as shown in (c) of FIG. 9, (i) a cap or valve connected to both ends of the main channel microtubule is opened, and (ii) a cap end connected to the outside of the plurality of subchannel microtubules. Or close with a valve. A washing solution for injecting a sample remaining in the microchannel for the main channel without reacting with the reactant is injected through either one of both ends of the main tube for the microchannel. As a result, as shown in Figure 9 (d), it is possible to remove the residual material through the washing liquid. Removing the residual material through such a washing solution, according to an embodiment, may be performed when the microfluidic analysis chip is manufactured, or by the user when the user uses the manufactured microfluidic analysis chip.

The washing liquid can be variously selected by the user according to the environmental conditions of the use of the micro-tubules for the main channel, and will generally be DIW (deionized water), PBS (phosphate buffered saline) or TBS (tris buffered saline) widely used in the bio field. Can be.

1 to 8 were all examples of fixing the reactant to the surface of the microchannel for the main channel. However, according to the present specification, the hydrogel may be fixed between the first point and the second point.

According to another embodiment of the present disclosure, (i) a first point of the first sub-channel microtube and the main channel microtube among the plurality of subchannel microtubes in the entire region of the main channel microtube; And (ii) the hydrogel 160 may be fixed to a surface of a region between a second point of the plurality of subchannel microtubes connected to the second subchannel microtube and the main channel microtube. In this case, the method may further include performing a surface treatment for fixing the hydrogel on the surface of the microchannel for the main channel.

The 'hydrogel' is a polymer material, which is generally used in diapers, contact lenses, medical electrodes, and cell cultures, and is used in a variety of bandages for molding materials, soil moisture storage, and burn wounds for special purposes. It is a hydrophilic polymer that is crosslinked by cohesive force such as covalent bond, hydrogen bond, van der waals bond or physical bond, and has a three-dimensional polymer network structure that can swell by containing a large amount of water in an aqueous solution. It is a substance having. For example, in order to form a concentration gradient of the chemicals by culturing the cells in three dimensions or diffusion of specific chemicals through the three-dimensional skeleton, Matrigel, Puratrix, Collagen, Fibrin gel, PEGDA, alginate, and the like. In addition, depending on the characteristics, the hydrogel formed by the Ionic cross-linking method has alginate (containing Ca2 + ions together), and PEGDA (initiator material) is used for the UV curable gel (requires photo-polymerization). And temperature sensitive gels such as collagen and matrigel. Since the type of the hydrogel is well known to those skilled in the art, a detailed description thereof will be omitted.

On the other hand, the hydrogel may be a reactant itself according to the present specification, it may be a medium containing a reactant according to the present specification. In addition, after the reactant according to the present specification is fixed to the surface of the microchannel for the main channel, the hydrogel may be injected to fill the microchannel for the main channel.

On the other hand, according to the present specification in the case of the first method of combining the chip upper plate and the chip lower plate, compared with the conventional manufacturing method, the chip upper plate before fixing the reaction material used as a reagent on the surface of the microchannel for the main channel The main difference is that the chip and the bottom plate are combined first. In the conventional manufacturing method, since the microchannel for the main channel of the microfluidic analysis chip is very small, when the reactant is a protein, the protein is first fixed on the surface of the microtubule, and then the upper chip plate and the lower chip plate are combined. At this time, since the heat treatment, ultraviolet treatment, and chemical treatment are used in the process of bonding the upper chip and the lower chip, the protein may be modified. Protein structure denaturation can lead to a decrease in analytical performance, which is a limitation for microfluidic analysis chips depending on the protein's characteristics. On the other hand, the microfluidic analysis chip 100 according to the present specification binds the chip upper plate 112 and the chip lower plate 111 first, and then fixes the protein on the surface of the microchannel for the main channel. Very unlikely.

On the other hand, the microfluidic analysis chip 100 according to the present specification is based on at least one of the measured impedance, the magnetic field and the optical value for the target region (i) whether the sample or reagent, (ii) the sample Or (iii) connecting a control unit for determining at least one of the flow rate of the reagent, (iii) the amount of the sample or the reagent, and (iv) the type of the sample or the reagent to the main channel microtubule. It may include.

The control unit may include a plurality of electrodes installed at both ends of a target region of the main channel microtubule and a sensor for measuring impedance between the plurality of electrodes for measuring impedance change.

The control unit may include a magnetic field measuring sensor installed at both ends of a target region of the microchannel for the main channel to measure the magnetic field change.

The control unit may include a light source installed at one end of the target region of the main channel microtubule and an optical sensor installed at the other end of the target region of the main channel microtubule for measuring optical numerical change.

10 is an enlarged view of a portion of a microfluidic analysis chip having a plurality of electrodes according to the present specification.

Referring to FIG. 10, it can be seen that two electrodes are installed in some regions of the microtube, and a voltage sensor for impedance measurement is connected between the two electrodes. Since gas has infinite impedance and liquid has relatively close impedance, it is possible to electrically measure the arrival of liquid on the region of interest when liquid and gas are injected in series, and thus the liquid to be checked in real time. Injection information can be used as accurate feedback control. In addition to the change in impedance, by adding a substance that affects the magnetic field to the sample or reagent, the magnetic field is changed to determine whether the liquid reaches the specific region, that is, the target region. In addition, by measuring the amount of light scattered in the process of the light emitted from the light source to pass through the sample or the reagent or by measuring the reflected light can also determine whether the liquid reaches the target area. This allows the control unit to (i) whether the sample or reagent reaches the target region based on the measured impedance, magnetic field or optical numerical value, (ii) the flow rate of the sample or the reagent, (iii) the sample or At least one of the amount of the reagent and (iv) the sample or the kind of the reagent can be determined.

When multiple samples and reagents were sequentially injected into the microtubules, the pre-injection order and volume information were previously stored and performed in anticipation of the passage time of a specific region according to the flow rate of the applied fluid. In this case, because of the pre-determined setting, it is impossible to know exactly when the samples and reagents are actually passed, and abnormal fluid drive may occur when an unexpected situation occurs or an error occurs in the preset. In contrast, the microfluidic analysis chip including the control unit according to the present specification includes (i) whether the sample or the reagent is reached, (ii) the flow rate of the sample or the reagent, (iii) the amount of the sample or the reagent, and (i) iv) The kind of the sample or the reagent can be determined. Thus, especially when performing biological assays, it is possible to determine when the samples and reagents arrive and to control subsequent fluid drive.

On the other hand, when a plurality of liquids having similar electrical conductivity are injected, there may be a restriction in distinguishing the liquid by the impedance difference. In this case, it is possible to monitor indirectly using index liquids with significantly different conductivity. That is, the flow velocity and the time point of passage can be determined based on the impedance information of the index liquid. This method is similarly applicable to magnetic field or optical numerical measurement methods.

According to the present specification, a plurality of subchannel microtubule microfluidic analysis chips 100 may include a cap or a valve that blocks fluid flow in the subchannel microtubules in some of the subchannel microtubules. Can be. In this case, the controller may control the opening and closing of at least one of the cap or the valve for blocking the fluid flow based on any one of the measured impedance, the magnetic field and the optical value.

11 and 12 are exemplary views of a microchannel for a subchannel including a valve according to an exemplary embodiment of the present specification.

Referring to FIG. 11, electrodes and sensors for measuring impedance are installed in the microchannel for the horizontal main channel. The valve is provided in the microchannel for longitudinal subchannels. (a) shows that the main channel microtubule is filled with gas, and the valve of the subchannel microtubule for liquid injection is closed. Then, as shown in (b), the injection of the subchannel is opened by starting the injection of the liquid when the injection of the liquid. Then, the liquid reaches between the electrodes as shown in (c). When the impedance between electrodes is measured, the gas has infinite impedance until before, but since the liquid reaches the zero impedance value, the liquid reaches the region of interest, and the flow rate can be monitored. In (d), after the target amount of liquid has been injected, the valve is closed to limit the inflow.

Referring to FIG. 12, in (a), pink liquid flows through the main channel microtubule, and the valve of the subchannel microtubule is closed. In the next (b), the injection of the blue tube by opening the valve of the subchannel when the injection of the blue liquid is required. In (c), by measuring the impedance between the electrodes, it is possible to monitor whether the blue liquid reaches the ROI, flow rate, and the like. Finally, in (d), after the target amount is injected, the valve is closed to limit the inflow.

While the embodiments of the present disclosure have been described with reference to the accompanying drawings, a person skilled in the art to which the present disclosure belongs may practice the present disclosure in other specific forms without changing the technical spirit or essential features. I can understand that. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive.

100: microfluidic analysis chip
110: Chip Housing
111: lower chip plate
112: upper chip plate
120: microchannel for the main channel
130: microchannel for subchannel
140: cap
150: protein fixing material
151: Protein
160: hydrogel

Claims (11)

(a) manufacturing a chip housing in which a microtube for the main channel and a plurality of microchannels for the subchannel are formed;
(b) performing a surface treatment for fixing a reactant on the surface of the microchannel for the main channel; And
(c) injecting a reactant through the microchannel for the main channel or the microchannel for the subchannel;
In step (b),
Caps which are connected to the outside of both ends of the main channel microtubules, the subchannel microtubules except for the first subchannel microtubules and the second subchannel microtubules among the plurality of subchannel microtubules, or Closing with a valve; And
(I) a first point to which the first subchannel microtube and the main channel microtube are connected; and (ii) the second subchannel microtube and the main channel. Injecting a surface treatment solution through the first subchannel microtubule or the second subchannel microtubule for surface treatment for fixing a reactant on the surface of the region between the second points to which the microtubules are connected;
Microfluidic analysis chip manufacturing method comprising a. The method according to claim 1,
In step (a),
Manufacturing a chip bottom plate and a chip top plate on which a main tube microtube and a plurality of subchannel microtubes are formed; And
Combining the lower chip and the upper chip plate; microfluidic analysis chip manufacturing method comprising a. The method according to claim 2,
The combining step, the microfluidic analysis chip manufacturing method, characterized in that for coupling the lower chip and the upper chip plate using at least one method of heat treatment, ultraviolet treatment and chemical treatment. The method according to claim 1,
The step (a) further comprises the step of forming a cap or valve for blocking the outside and the main tube micro-tubes or sub-channel micro-tubes on the main channel micro-pipes or sub-channel micro-pipes; Microfluidic analysis chip manufacturing method characterized in that. delete (a) manufacturing a chip housing in which a microtube for the main channel and a plurality of microchannels for the subchannel are formed;
(b) performing a surface treatment for fixing a reactant on the surface of the microchannel for the main channel; And
(c) injecting a reactant through the microchannel for the main channel or the microchannel for the subchannel;
Including,
In step (c),
Closing both ends of the main channel microtubules and the ends connected to the outside of the other subchannel microtubules except for the first and second subchannel microtubules with a cap or a valve; And
(I) a first point to which the first subchannel microtube and the main channel microtube are connected; and (ii) the second subchannel microtube and the main channel. Injecting a reactant solution through the first subchannel microtube or the second subchannel microtube to fix the reactant to the surface of the region between the second points to which the microtubules are connected; Microfluidic analysis chip manufacturing method. (a) manufacturing a chip housing in which a microtube for the main channel and a plurality of microchannels for the subchannel are formed;
(b) performing a surface treatment for fixing a reactant on the surface of the microchannel for the main channel; And
(c) injecting a reactant through the microchannel for the main channel or the microchannel for the subchannel;
Including,
(d) opening the cap or valve of (i) the cap or valve of the first or second subchannel microchannel and (ii) the cap or valve of the main channel microtubule adjacent to the cap or valve selected in (i) above; And
(e) injecting a removal solution for removing a non-fixed reactant on the surface of the region between the first point and the second point through the subchannel microtube or the main channel microtube opened in (d); Microfluidic analysis chip manufacturing method characterized in that it further comprises. (a) one end is connected to the main channel microtube providing a space for reacting with the reagent while the sample introduced from the formed sample inlet is moved to the other end, one end is connected to the side of the main channel microtube, the other end is A microfluidic analysis chip comprising a plurality of subchannel microtubes connected to an outside of the chip housing, and a chip housing surrounding the main channel microtubes and the plurality of subchannel microtubes.
(I) a first point at which a first subchannel microtube and the main channel microtube are connected among the plurality of subchannel microtubes, and (ii) the plurality of subchannels. A sample is injected through the sample inlet of the microfluidic analysis chip in which a reactant or a hydrogel is fixed on a surface of a region between a second subchannel microtube and a second point to which the main channel microtube is connected. Doing;
(b) (i) opening the cap or valve connected to both ends of the microchannel for the main channel, and (ii) closing the ends connected to the outside of the plurality of subchannel microtubes with a cap or valve; And
(c) injecting a washing solution for washing the sample remaining in the microchannel for the main channel without reacting with the reactant through one of both ends of the main tube for the microchannel; How to use a fluid analysis chip. (a) manufacturing a chip housing in which a microtube for the main channel and a plurality of microchannels for the subchannel are formed;
(b) closing both ends of the main tube microtubules and the ends connected to the outside of the remaining microchannel microtubes except for the first and second subchannel microtubes with a cap or a valve; And
(c) injecting a hydrogel through the first or second subchannel microtubules. The method according to claim 9,
(b-1) surface treatment for fixing the hydrogel on the surface of the microchannel for the main channel; microfluidic analysis chip manufacturing method further comprising. The method according to claim 1,
(d) whether or not the sample or reagent is reached, (ii) the flow rate of the sample or the reagent, based on at least one of the measured impedance, magnetic field and optical value for the target region, (iii) the sample Or (iv) connecting a control unit for determining at least one of the amount of the reagent and (iv) the sample or the type of the reagent to the main channel microtubules. How to make.

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