KR101750041B1 - Microfluidics chip - Google Patents

Abstract

The microfluidics chip has a main plate on which a series of concentration gradient forming channels are formed and two or more reagent supply channels for supplying reagents to the concentration gradient forming channels and is stacked on the main plate, , And the concentration of the reagent supplied through the reagent supply channel to the concentration gradient forming channel can be linearly graded in the concentration gradient forming channel.

Description

[0001] MICROFLUIDICS CHIP [0002]

The present invention relates to a microfluidic chip capable of mixing a reagent and capable of a concentration gradient with respect to a reagent.

Bio chip is a biological microchip capable of analyzing gene expression patterns, gene defects, protein distribution, and reaction patterns by combining biological molecules such as DNA and protein on a small substrate made of glass, silicon or nylon. (Biological Microchip).

The biochip can be divided into a microarray and a microfluidics chip. A microarray arranges thousands or tens of thousands of DNAs or proteins at regular intervals, processes the substance to be analyzed, The microfluidics chip is also referred to as a lab-on-a-chip (LOC), which reacts with various biomolecules or sensors that are concentrated on the chip while flowing a small amount of analyte. The biochip can be analyzed. Recently, microfluidics chips have been expanding the field of separation, synthesis and quantitative analysis of analytes.

Until now, in the case of using a cell culture plate in an experimental procedure, it was common to manually fill each well in a cell culture plate with a micropipette. This method is a simple repetitive operation which is not efficient in terms of time or manpower and is not suitable for preparation of reagents at various concentrations.

In practice, it is necessary to confirm the response to various concentrations of reagents such as toxicity evaluation for drug development, mixing of cell culture medium in the field of medical and biochemical experiments, toxicity test according to mixing ratio of toxic substance, biochemical experiment, Conventional microfluidics chips have limitations in the above experiments.

For reference, Korean Patent Laid-Open Publication No. 2003-0008340 discloses a lab-on-a-chip for analyzing an amine compound. However, even in the above-mentioned patent, the reaction to the analyte can be confirmed only for one drug at a time.

The present invention provides a microfluidic chip capable of mixing various reagents and enabling a concentration gradient with respect to a reagent to confirm various reactions at once.

The present invention is not limited to a linear concentration gradient of a reagent but provides a microfluidics chip capable of three-dimensionally grading a reagent concentration.

According to a preferred embodiment of the present invention for achieving the objects of the present invention, a microfluidic chip capable of mixing a plurality of reagents includes a main plate on which a series of concentration gradient forming channels are formed; And a laminating plate on which two or more reagent supply channels for supplying the reagent to the concentration gradient forming channel are formed and which are stacked on the main plate and to which a concentration gradient forming channel and a reagent supply channel are connected, The concentration of the reagent supplied to the channel can be linearly graded in the concentration gradient forming channel.

A plurality of reagents can be mixed in one concentration gradient forming channel and the concentration gradient for the plurality of reagents is linearly formed along the concentration gradient forming channel so that the concentration of the reactant Can be confirmed.

Meanwhile, in the microfluidic chip according to the present invention, the concentration gradient forming channel and the reagent supplying channel can form a height difference at the intersection point where they are connected to each other, so that the reagent can be formed simply in two dimensions along the concentration gradient forming channel It is possible to more accurately confirm the reaction between the reagent and the reaction object, as compared with the case where it is supplied.

For example, the height difference may cause the reagent to flow in a direction perpendicular to the longitudinal direction of the concentration gradient forming channel at the intersection point, and the reaction object to be disposed at the intersection point by the reagent flowing in three dimensions, The whole can react completely to the reagent.

As an example of forming the height difference between the concentration gradient forming channel and the reagent supply channel, a concentration gradient forming channel may be formed on one surface of the body plate, a reagent supply channel may be formed on one surface of the lamination plate, The plates can be connected to each other to connect the two channels with a height difference.

In addition, the buffer solution can be introduced into the concentration gradient forming channel so that the reagent moves with a direction in the concentration gradient forming channel, and the reagent can move along the moving direction of the buffer solution.

For example, when the buffer solution is injected from one end of the concentration gradient forming channel to the other end, the reagent also moves from one end to the other end of the concentration gradient forming channel along the moving direction of the buffer solution. Alternatively, the buffer solution may be injected at the center of the concentration gradient forming channel, in which case the buffer solution may migrate from the center of the concentration gradient forming channel to either end and the reagent may also flow along the buffer solution at the center of the concentration gradient forming channel Move to both ends. In this case, by injecting the same reagents to the left and right with respect to the center of the concentration gradient forming channel, the reaction of the reaction object to the same reagent can be confirmed again in one concentration gradient forming channel.

In addition, the concentration gradient forming channel may be formed by a combination of at least one of a straight line, a curve, and a zigzag. By forming a concentration gradient forming channel in a curved or zigzag shape, A combination of many reagents and concentration gradients can be realized, which allows the reaction of the reaction object to be confirmed with more reagents.

Further, the adjustment of the concentration gradient of the reagent in the concentration gradient forming channel can be determined by any one of the number and width of the reagent supply channel, the width of the concentration gradient forming channel, and the flow rate of the reagent.

In addition, two or more lamination plates may be laminated on the body plate, so that the response of the reaction object to more reagents can be confirmed. For example, two lamination plates may be stacked on the top and bottom surfaces, respectively, of the body plate, in which case the concentration gradient forming channels are provided in a hole shape through the body plate such that the reagent supply channels are in the Connect them from top to bottom.

In the conventional microfluidics chip, there is a limitation in confirming the response of a reaction object to one reagent, which is a problem in confirming the response to various reagents. In the microfluidic chip of the present invention, however, The reagent supply channel can be connected to confirm the response to a plurality of reagents.

Also, by connecting the concentration gradient forming channel and the reagent supply channel at a height difference, the reagent can be introduced in a direction perpendicular to the longitudinal direction of the concentration gradient forming channel, and the reagent is two-dimensionally The reagent is supplied three-dimensionally, so that the reaction between the reagent and the reaction object can be realized more accurately at the crossing point.

In addition, although the concentration gradient forming channel may be provided as a straight line, it may be provided by a curve or a zigzag, so that a combination of the reagents and a concentration gradient can be realized in the same area of the microfluidics chip, Reaction confirmation is possible.

Further, by laminating a plurality of lamination plates on the body plate on which the concentration gradient forming channels are formed, it is possible to confirm the reaction of the reaction object to more reagents.

1 is a perspective view of a microfluidic chip according to an embodiment of the present invention.
2 is an exploded perspective view of a microfluidic chip.
FIG. 3 is a view for explaining that a concentration gradient forming channel and a reagent supply channel are connected with a height difference, and a reagent reacts to a reaction object three-dimensionally.
4 is a view for explaining another method of injecting the buffer solution.
Figure 5 shows microfluidics chips that provide the concentration gradient forming channels in straight, zigzag, and curved lines.
6 and 7 are views of a microfluidic chip having a plurality of lamination plates.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the present invention is not limited to the embodiments. For reference, the same numbers in this description refer to substantially the same elements and can be described with reference to the contents described in the other drawings under the above-mentioned rules, and the contents which are judged to be obvious to the person skilled in the art or repeated can be omitted.

1 is a perspective view of a microfluidic chip according to an embodiment of the present invention. FIG. 2 is an exploded perspective view of a microfluidic chip, and FIG. 3 is a view illustrating a three-dimensional reaction of a reagent with a concentration gradient forming channel and a reagent supply channel connected to each other with a height difference.

1 to 3, a microfluidic chip 100 according to the present embodiment has a body plate 110 and a lamination plate 120.

The plates may be made of a material such as glass, silicon, nylon or the like, and it is preferable to use a material which is less reactive with acid, base and other biochemical substances. Particularly, polydimethylsiloxane (PDMS) can be used as the material of the body plate and the lamination plate, and polydimethylsiloxane and silicon PMMA (poly methyl methacrylate) and the like are also possible. The material is a polymer material capable of mutual adhesion, and a stable close contact state between the plates can be realized. As will be described later in detail, the plate is fully covered with the channel provided in a groove shape to effectively maintain the closed state of the channel.

The concentration gradient forming channel 112 is formed in a groove shape on the upper surface of the body plate 110. In the present embodiment,

At both ends of the concentration gradient forming channel 112, a buffer space 111 capable of accommodating a reagent or a buffer solution is provided.

Four reagent supply channels 122 for injecting reagents are formed in the lamination plate 120. In the present embodiment, four reagent supply channels 122 are provided, It can be changed according to the kind of the reagent or the concentration gradient. The reagent supply channel 122 is provided in a groove shape on the bottom surface of the lamination plate 120.

A reagent supply hole 121 for injecting a reagent into the reagent supply channel 122 may be formed in the lamination plate 120. A reagent supply hole 121 may be connected to an end of the reagent supply channel 122, A reagent hose 123 for injection can be connected. In addition, a buffer solution supply hole 124 for supplying a buffer solution is formed in the lamination plate 120.

When the two plates are bonded to each other, the concentration gradient forming channel 112 of the body plate 110 and the reagent supply channel 122 of the laminating plate 120 can be connected. The buffer solution supply hole 124 is connected to the buffer space 111 provided on the left side of the body plate 110 and the buffer solution hose 125 for supplying the buffer solution is connected to the buffer solution supply hole 124 . In the present embodiment, a buffer solution supply hole 124 is provided on both sides of the lamination plate 120, and a buffer solution hose 124 can be connected to either side. However, in this embodiment, the buffer solution hose 125 is connected to the buffer solution supply hole 124 on the left side, and the buffer solution moves from left to right along the concentration gradient forming channel 112. Therefore, in the present embodiment, the buffer solution supply hole 124 on the right side can be used as a passage through which the reagent and the buffer solution are discharged. In some cases, the buffer solution supply hole 124 is filled with the buffer solution hose 124 And the direction of movement of the buffer solution can be switched.

The concentration gradient forming channel 112 and the reagent supply channel 122 except the portion where the concentration gradient forming channel 112 and the reagent supply channel 122 intersect with each other are in close contact with each other And can be covered by the plates.

In this embodiment, the buffer solution is injected to control the moving direction and speed of the reagent flowing into the concentration gradient forming channel 112 through the reagent supply channel 122. In some cases, the buffer solution may be omitted You may. Further, the buffer solution may be changed depending on the kind of the reaction object or the reagent, and the buffer solution itself is preferably selected as a liquid substance which does not react with the reaction object or the reagent. In the present embodiment, the buffer solution and the reagent may include all materials capable of flowing. The buffer solution and the reagent may be replaced by a culture medium of the cells, and the reaction target may also be a drug EGF (epithelial growth factor) In addition, it is possible to check the concentration gradient of doxorubicin (doxorubicin), which can test for toxicity.

For reference, the loading of the buffer solution or the reagent into the concentration gradient channel may be performed by adjusting the injection pressure of the buffer solution or the reagent. Alternatively, a separate suction provided at the opposite end of the concentration gradient channel into which the buffer solution is injected and may implement loading of the liquids through suction. However, in such a case, there is a limitation in the loading operation of the buffer solution or the reagent, and it is preferable to use the self-weight of the solutions or the compressors to individually inject the solutions. By the method of injecting the buffer solution or the reagent But is not limited to.

In the microfluidic chip 100 according to the present embodiment, at least one to four reagents may be mixed in one concentration gradient forming channel 112, and a concentration gradient for the plurality of reagents may be mixed with the concentration gradient forming channel 112, The various reactions of the reaction object depending on the kind and concentration of the reagent can be confirmed on the concentration gradient forming channel 112. [

For reference, in this embodiment, the width of the reagent supply channel and the concentration gradient forming channel is set to 100 μm, and the flow rate of the reagent or the buffer solution is adjusted to 0.1 mL / min in order to realize an optimized concentration gradient. However, the present invention does not limit the width or flow rate of the channel to the above numerical values. This is a matter that can be changed depending on the viscosity of the reagent or the buffer solution or the characteristics of the material of the plates, and the simulation of the manufactured microfluidic chip Can be changed appropriately. However, it has been confirmed that the concentration gradient at the crossing point can be well realized as the number of the reagent supply channels is increased in the microfluidics chip manufactured as described above. However, the concentration gradient at the crossing point The microfluidics chip having four reagent supply channels was fabricated as the most ideal structure because the tendency that the difference becomes too small occurs.

Also, the types of reagents provided to the reagent supply channel may be the same or different. For example, when the reagents are provided in the same kind, the reaction of the reaction object can be confirmed for reagents having different concentrations at each crossing point, and analysis of the reagent itself is possible. In addition, when different kinds of reagents are supplied to the reagent supply channel, it is possible to analyze the reaction of the reaction object and the reagent itself to reagents mixed at different ratios. That is, through the microfluidic chip according to the present invention, various reagents can be analyzed and various reactions of the reaction object to the reagents can be confirmed. However, depending on the kind of the reagent and the change of the reaction object, The present invention is not limited thereto.

Meanwhile, in the microfluidic chip 100 according to the present invention, the concentration gradient forming channel 112 and the reagent supply channel 122 may form a height difference at intersections 130 connected to each other, It is possible to more accurately confirm the reaction between the reagent and the reaction object, as compared with the case where the reaction solution is simply supplied two-dimensionally along the concentration gradient forming channel.

Specifically, when the reagent flows into one passage as in the conventional method, the reaction between the reagent and the reaction object may not occur at a portion where the reaction object is not completely immersed in the reagent, May not fully react with the reagent.

However, in the microfluidic chip 100 according to the present invention, the above-mentioned problems can be solved by connecting the reagent supply channel 122 into which the reagent is injected and the concentration gradient forming channel 112 in an overlapping manner.

Specifically, in this embodiment, since the concentration gradient forming channel 112 and the reagent supply channel 122 are provided on separate plates and are joined to connect the channels, the two channels can be naturally arranged up and down . 3, the height difference of the two channels allows the reagent to flow in the direction 134 perpendicular to the longitudinal direction 132 of the concentration gradient forming channel 112 at the intersection point 130, The reaction object 10 disposed at the crossing point 130 by the reagent flowing in the right and left directions, that is, in three dimensions, can fully react with the reagent.

As described above, the buffer solution is for controlling the moving direction of the reagent flowing into the concentration gradient forming channel. In the foregoing embodiment, the buffer solution flows from the left side to the right side in the drawing of FIGS. 1 to 3, The flow from left to right along the buffer solution is described as an example. In the examples described below, the buffer solution moves from the center of the concentration gradient channel to both ends.

Referring to FIG. 4, the body plate 110 and the lamination plate 120 of the microfluidics chip 100 of this embodiment are substantially the same as those of the microplicity chips described above. In this embodiment, And the reference numerals for substantially the same components are the same as those of the previous embodiment.

A buffer solution supply hole 124 for injecting the buffer solution in this embodiment is formed in the lamination plate 120 and the buffer solution supply hole 124 is formed through a concentration gradient Forming channel 112. In some cases, however, the buffer solution supply holes may be adjusted in position so that they are directly connected to the concentration gradient forming channel in the process of bonding the body plate and the lamination plate, which are disposed up and down.

Referring to FIG. 4, the buffer solution is introduced into the center of the concentration gradient forming channel 112, and the buffer solution can move toward both ends of the concentration gradient forming channel 112. The reagent also migrates from the center of the concentration gradient channel 112 along the buffer solution to both ends.

In this case, by injecting the same reagent into the reagent supply channel 122, which is disposed symmetrically with respect to the center of the concentration gradient forming channel 112, the concentration of the reactant to the same reagent in one concentration gradient forming channel 112 You can see the reaction twice. Specifically, in this embodiment, a reagent supply channel 122 capable of supplying three reagents A, B, and C is connected to the right and left sides of the center of the concentration gradient forming channel 112, You can repeatedly check for the reaction.

For reference, the lamination plate 120 may have a discharge hole 127 connected to both ends of the concentration gradient forming channel 112, and the reagent and the buffer solution may be discharged through the discharge hole 127.

FIG. 5 shows a microfluidic chip having density gradient forming channels of various shapes.

5 (a) shows a linear concentration gradient forming channel 112, and four reagent supply channels 122 are connected. 5 (b) shows a zigzag concentration gradient forming channel 112, and eight reagent supply channels 122 are connected. 5 (c) shows a curved density gradient forming channel 112, and ten reagent supply channels 122 are connected.

As can be seen from the figure, more concentration and concentration gradient can be realized by providing concentration gradient channels of curve or zigzag shape rather than a straight line in microfluidics chips having the same area, The reaction of the reaction object can be confirmed.

6 and 7 show a microfluidic chip having a plurality of lamination plates. Referring to FIG. 1, a lamination plate 120 is stacked on a body plate 110 so that reaction of a reaction object against more reagents can be confirmed. Specifically, the concentration gradient forming channel 112 is provided in a hole shape passing through the body plate 110, and the reagent supply channel 122 of the lamination plate 120, which is stacked on the body plate 110, Forming channel 112. In this way,

The concentration gradient forming channel 112 excluding the portion where the concentration gradient forming channel 112 and the reagent supplying channel 122 intersect with each other is formed while the two lamination plates 120 are bonded above and below the body plate 110. [ And the reagent supply channel 122 can be covered by the plates that are in close contact with each other, thereby preventing the reagent or the buffer solution from splashing out of the channel.

In Fig. 7, the moving direction of the reagent is shown from left to right, but this depends on the moving direction of the buffer solution, and the direction can be switched.

Although the present invention has been described with reference to the preferred embodiments thereof, it will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the invention as defined in the following claims. It can be understood that

10: reaction object 100: microfluidic chip
111: buffer space 112: concentration gradient forming channel
120: Lamination plate 121: Reagent supply hole
123: Reagent hose 124: Buffer solution supply hole
125: buffer solution hose 130: intersection point

Claims (10)

In a microfluidic chip capable of mixing a plurality of reagents,
A body plate on which a series of concentration gradient forming channels are formed; And
And a laminating plate on which at least two reagent supply channels for supplying a reagent to the concentration gradient forming channel are formed and the concentration gradient forming channel and the reagent supply channel are connected to each other,
A plurality of lamination plates are stacked on the body plate,
Wherein the concentration gradient forming channel is provided in a hole shape passing through the body plate, the lamination plate is stacked on the upper surface and the lower surface of the body plate, the reagent supply channel is connected above and below the concentration gradient forming channel,
Wherein the concentration of the reagent supplied to the concentration gradient forming channel through the reagent supply channel is linearly graded in the concentration gradient forming channel. The method according to claim 1,
Wherein the concentration gradient forming channel and the reagent supply channel form a height difference at crossing points connected to each other,
And the reagent flows in a direction perpendicular to the longitudinal direction of the concentration gradient forming channel at the intersection point by the height difference. 3. The method of claim 2,
Wherein the concentration gradient forming channel and the reagent supply channel are provided as grooves formed on the body plate and the laminating plate, respectively. The method according to claim 1,
Wherein the buffer solution flows into the concentration gradient forming channel and the moving direction of the reagent flowing into the concentration gradient forming channel follows the moving direction of the buffer solution. 5. The method of claim 4,
Wherein the buffer solution moves from one end to the other end of the concentration gradient forming channel. 5. The method of claim 4,
Wherein the buffer solution moves from the center of the concentration gradient channel to both ends. The method according to claim 1,
Wherein the concentration gradient forming channel is formed of a combination of at least one of a straight line, a curve, and a zigzag. The method according to claim 1,
The adjustment of the concentration gradient of the reagent in the concentration gradient forming channel can be accomplished,
The number and width of the reagent supply channel, the width of the concentration gradient forming channel, and the flow rate of the reagent. ≪ IMAGE > delete delete

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