KR102201719B1 - Lab-on-a-chip System using Microfluid Control based on the Finger Actuation and Uses Thereof - Google Patents

Abstract

The present invention relates to a lab-on-a-chip system using finger-driven microfluidic control and its use, and in more detail, microfluids can be easily controlled through the operation of pneumatic valves and pneumatic chambers using pressure generated by finger driving. Regarding the lab-on-a-chip system, the lab-on-a-chip system according to the present invention can simply drive the lab-on-a-chip system by moving microfluids with pressure according to finger driving without being affected by the operating characteristics of the user's fingers. In addition, since the pressure chamber and the microfluidic moving part are separated from each other, the microfluidic channel can be integrated regardless of the position of the pressure chamber, which is useful for multi-fluid analysis.

Description

Lab-on-a-chip System using Microfluid Control based on the Finger Actuation and Uses Thereof}

The present invention relates to a lab-on-a-chip system using microfluidic control and a use thereof, and more particularly, to a lab-on-a-chip system that can easily control a microfluid through the operation of a pneumatic valve and an operation chamber using external pressure. .

The lab-on-a-chip system using microfluidic control technology implements the functions of bulky and expensive equipment mainly used in laboratories on a chip the size of a finger, so that various pre-processing and analysis required in the laboratory can be efficiently performed within a short time. (Curtis et al., Lab Chip. Vol. 7, pp. 41-57, 2007). However, in order to drive a general lab-on-a-chip system, the method of use is complex and requires an expensive fluid driven pump. For this reason, a lab-on-a-chip system having numerous advantages has not been widely used.

In order to solve the problems of the lab-on-a-chip system, a lab-on-a-chip system is implemented by using a capillary phenomenon of paper fibers and microstructures, or by driving a microfluid using a gas removal process of a porous material, PDMS (polydimethylsiloxane). I did. However, there is a limitation in that the analysis time using the lab-on-a-chip system is lengthened due to the slow driving speed of the microfluid. In addition, the analysis using large-sized cells is limited due to the paper fibers and microstructure.

On the other hand, various lab-on-a-chip systems such as glucose test, blood type test, antibiotic resistance test, microdrop generation, etc. have been reported through microfluidic drive using finger operation, which is one of the simplest actions that humans can take (US 2016- 0109467, Xu K et al., Lab Chip., Vol. 15, pp. 867-76, 2015, Glynn MT et al., Lab Chip. Vol. 14, pp. 2844-51, 2014, KR10-2013-0058654A , US2016-0144362A).

In the above report, the microfluidic is temporarily driven by simply applying pressure in the microfluidic channel with a finger operation, and the continuous microfluidic operation is also possible using pneumatic valves. Although various lab-on-a-chip systems were implemented by simply driving the microfluid in this way, there is a disadvantage that the characteristics of the microfluid are affected by the various finger operation characteristics of users because it is difficult to precisely control through finger operation.

That is, there is a limitation in providing elaborate and standardized guidelines in consideration of various finger operation characteristics of users. In addition, since the pressure chamber, which applies pressure using finger operation, is directly connected to the microfluidic channel, the position of the pressure chamber must be considered when designing a lab-on-a-chip system through integration of microfluidic channels. Therefore, there is a limitation in that the integration of microfluidic channels is limited. Additionally, due to the fluid flowing into the pressure chamber, it is possible to influence the operation of the fingers.

Accordingly, the present inventors have made diligent efforts to develop a lab-on-a-chip system that can move microfluids at a uniform pressure without being affected by finger operation characteristics. As a result, the moving part and the pneumatic channel through which the fluid moves are separated into a polymer thin film. In this case, not only can the microfluid be moved without being affected by the finger operation characteristics, but the microfluidic channel is not directly connected to the pressure chamber, so the microfluidic channel does not flow back into the pressure chamber, so that the microfluidic channel can be integrated in various forms. It was confirmed that it was possible, and the present invention was completed.

The information described in the background section is only for improving an understanding of the background of the present invention, and thus does not include information forming the prior art known to those of ordinary skill in the art to which the present invention belongs. May not.

An object of the present invention is to provide a wrap-on-a-chip system using a finger-driven microfluidic control that is not affected by the user's finger operation characteristics.

Another object of the present invention is to provide a method for controlling microfluids using the system.

Another object of the present invention is to provide a lab-on-a-chip system for analyzing multiple fluids including the system as one fluid drive unit.

In order to achieve the above object, the present invention is a moving unit 1200 to move the fluid; A pressure chamber 1100 driven by external pressure; A pneumatic channel 1000 through which compressed air moves; An injection unit 100 for injecting a fluid; A discharge unit 200 for discharging the fluid; And a pneumatic discharge unit 300, wherein the moving unit and the pneumatic channel are separated by a polymer thin film 900, and a lab-on-a-chip using microfluidic control is provided.

The present invention also includes the steps of: (a) applying an external pressure to the pressure chamber of the lab-on-a-chip; (b) blocking the pneumatic discharge unit while external pressure is applied; (c) removing the external pressure applied in step (a); (d) injecting the sample into the injection unit; And (e) applying and removing external pressure to the pressure chamber; and a method for controlling microfluids of a lab-on-a-chip.

The present invention also provides a lab-on-a-chip using microfluidic control for multi-fluid analysis, including the lab-on-a-chip as a fluid driving unit for driving one type of fluid, and a plurality of the fluid driving units are connected.

The lab-on-a-chip system according to the present invention can not only drive the lab-on-a-chip system simply by moving microfluids under the pressure of the finger drive, but also the pressure chamber and microfluidic system without being affected by the operating characteristics of the user's fingers. Since the fluid moving parts are separated from each other, microfluidic channels can be integrated regardless of the position of the pressure chamber, which is useful for multi-fluid analysis.

1 is a schematic diagram showing the structure of a lab-on-a-chip system according to an embodiment of the present invention.
2 shows a component diagram of a lab-on-a-chip system according to an embodiment of the present invention.
3 is a schematic diagram that simulates fluid movement according to finger driving of the lab-on-a-chip system according to an embodiment of the present invention, and the left panel shows the state of the lab-on-a-chip and fluid when pressure is applied to the pressure chamber by pressing the finger. It shows the movement, and the right panel shows the state of the wrap-on-a-chip and the movement of the fluid when the pressure in the pressure chamber is reduced by removing the finger.
4 is a flowchart of a microfluidic control method using finger driving in a lab-on-a-chip system according to an embodiment of the present invention.
5 is an evaluation of the driving characteristics of the lab-on-a-chip system according to an embodiment of the present invention, (A) is a graph measuring the amount of movement of the microfluid according to the pressed depth of the pressure chamber, (B) is the pressure When applied 4 times, it is a graph measuring the movement amount of the microfluid according to the time interval in which the pressure is applied, and (C) is a graph measuring the movement amount of the microfluid according to the four finger movements of several people.
6 is for the result of adjusting the amount of fluid in the lab-on-a-chip system according to an embodiment of the present invention, (A) is the amount of microfluid that has moved according to the diameter of the operating chamber and the pressure recovery applied to the pressure chamber. Is the measured graph, and (B) is a graph comparing the calculated volume of the operating chamber and the actually measured amount of fluid.
7 is a diagram showing a lab-on-a-chip system for sample mixing prepared as an example of application of a lab-on-a-chip system according to an embodiment of the present invention, and (A) is a schematic diagram of a lab-on-a-chip system for sample mixing (B) is the result of measuring the ratio of the mixed solution according to the diameter ratio of the operating chamber, and (C) is a graph comparing the predicted value and the actual measured value.
FIG. 8 shows a lab-on-a-chip system for detecting E. coli produced as an example of use of the lab-on-a-chip system according to an embodiment of the present invention. (A) is a substrate metabolized by an enzyme inside E. This change describes the process, (B) is a schematic diagram depicting a lab-on-a-chip system for detecting E. coli using four fluid drive units and two pressure chambers manufactured in an embodiment of the present invention, (C) Is the result of observing the color change of the reaction substrate by E. coli concentration in the reaction chamber, and (D) is a graph showing this.
9 is a lab-on-a-chip system for a blood cross-conformity test manufactured as an example of application of the lab-on-a-chip system according to an embodiment of the present invention, (A) using four fluid drive units and two pressure chambers. , A schematic diagram of a system that can separate the blood of a donor and a donor into plasma and blood cells, react in a reaction chamber, and observe the result, (B) is a lab-on-a-chip system that was actually produced, and (C) is It shows the part diagram of the part where the membrane separating blood cells and plasma is installed after injecting the blood of the recipient and the recipient.
FIG. 10 is a comparison between the results of testing the blood type combination of an actual patient using the system and the results using the conventional method, and it can be seen that the blood coagulation reactions are the same.
FIG. 11 is a lab-on-a-chip system for extracting nucleic acids produced as an application example of the lab-on-a-chip system according to an embodiment of the present invention, with 4 fluid driving units, 3 pressure chambers, and a reaction part containing glass beads It shows a lab-on-a-chip system for extracting nucleic acids containing.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by an expert skilled in the art to which the present invention belongs. In general, the nomenclature used in this specification is well known and commonly used in the art.

Terms such as 1st, 2nd, A, B, etc. may be used to describe various components, but the components are not limited by the above terms, only for the purpose of distinguishing one component from other components. Is only used. For example, without departing from the scope of the rights of the technology described below, a first component may be referred to as a second component, and similarly, a second component may be referred to as a first component. The term and/or includes a combination of a plurality of related listed items or any of a plurality of related listed items.

In terms of the terms used in the present specification, expressions in the singular should be understood as including plural expressions unless clearly interpreted differently in context, and terms such as "includes" are specified features, numbers, steps, actions, and components. It is to be understood that the presence or addition of one or more other features or numbers, step-acting components, parts or combinations thereof is not meant to imply the presence of, parts, or combinations thereof.

Prior to the detailed description of the drawings, it is intended to clarify that the division of the constituent parts in the present specification is merely divided by the main function that each constituent part is responsible for. That is, two or more constituent parts to be described below may be combined into one constituent part, or one constituent part may be divided into two or more according to more subdivided functions. In addition, each of the constituent units to be described below may additionally perform some or all of the functions of other constituent units in addition to its own main function, and some of the main functions of each constituent unit are different. It goes without saying that it may be performed exclusively by.

In addition, in performing the method or operation method, each of the processes constituting the method may occur differently from the specified order unless a specific order is clearly stated in the context. That is, each process may occur in the same order as the specified order, may be performed substantially simultaneously, or may be performed in the reverse order.

In the present invention, the efficiency of the lab-on-a-chip system was not affected by the finger driving characteristics and the pressure chamber and the microfluidic moving part were not directly connected.

In the present invention, the microfluidic moving unit; Operating chamber; Regulating valve; And when the polymer thin film is separated between the first substrate including the pressure chamber and the second substrate including the pneumatic channel, not only are not affected by the finger driving characteristics, but also the microfluidic moving part and the compressed air moving It was confirmed that the channels are not directly connected so that the microfluidic moving part can be freely integrated.

That is, in an embodiment of the present invention, an injection unit for injecting a fluid; First and second moving parts through which the fluid moves; First and second pneumatic valves for controlling the movement of fluid; An operation chamber for moving the fluid; A pressure chamber driven by a finger; Pneumatic outlet; And a first substrate including a discharge unit for discharging the fluid; A second substrate including a pneumatic channel through which compressed air moves; And in a lab-on-a-chip system including a PDMS thin film inserted between the first and second substrates, when the pneumatic channel is connected only to the first valve and the operation chamber, the movement of the microfluid is controlled without being affected by the finger driving characteristics. It was confirmed that it can be done (Fig. 1).

Accordingly, the present invention, in one aspect, the moving unit 1200 to move the fluid; A pressure chamber 1100 driven by external pressure; A pneumatic channel 1000 through which compressed air moves; An injection unit 100 for injecting a fluid; A discharge unit 200 for discharging the fluid; And a pneumatic discharge unit 300, wherein the moving unit and the pneumatic channel are separated by a polymer thin film 900. It relates to a lab-on-a-chip using microfluidic control.

In the present invention, the external pressure may be used without limitation as long as it is a method capable of applying pressure to the pressure chamber, but may be preferably a pressure pressed with a finger, but is not limited thereto.

In the present invention, the moving part first and second moving parts, first and second pneumatic valves for controlling the movement of the fluid; And an operation chamber for moving the fluid. It may be characterized in that it includes a.

In the present invention, the pressure chamber can be operated as long as it is present in a position where external pressure can be applied regardless of the position of the moving part or the pneumatic channel, and preferably it is present on the upper substrate of the lab-on-a-chip system. It can be characterized.

Hereinafter, the lab-on-a-chip system according to the present invention will be described in detail with reference to the drawings.

1 is a schematic diagram showing the most basic structure of the lab-on-a-chip system devised in the present invention, and shows a system capable of controlling the movement of microfluids by a single pressure chamber.

2 is a schematic diagram of the most basic structure of the lab-on-a-chip system devised in the present invention, and detailed descriptions of each component are as follows. In addition, as disclosed in FIG. 2, the lab-on-a-chip of the present invention is provided on/off the moving part and the pneumatic channel as long as the moving part moving the microfluid and the pneumatic channel moving the compressed air are separated by a polymer thin film. It is obvious to a person skilled in the art that the positions below can be interchanged.

The injection unit 100 is a site in which a sample is injected, and any sample surface that can be used for an analysis/separation system using microfluids can be used, and the injection unit may be characterized in that it further includes a sample separation membrane. However, it is not limited thereto. For example, when the lab-on-a-chip system of the present invention is used in a blood cross-conformance test, plasma and blood cells of a donor and a recipient must be separated. In this case, the injection unit 100 may further include a blood separation membrane to separate blood into plasma and blood cells before moving to the microfluidic moving unit.

The discharge unit 200 is used for discharging a sample that has been analyzed/separated, and in the simplest structure, it is used as a simple discharge unit, but in a multi-fluid analysis, the discharge unit may be used as an injection unit having a different structure.

The pneumatic discharge unit 300 discharges the air remaining in the pneumatic channel 1000 when the pressure chamber 1100 is pressed with a finger. When the lab-on-a-chip system of the present invention is first operated, the air is discharged. Then, it may be characterized in that it is closed during continuous operation to prevent additional air inflow.

The first pneumatic valve 400 and the first moving unit 500 control the step of moving to the operation chamber 600 after the microfluid is injected through the injection unit, and the first pneumatic valve and the operation chamber 600 may be directly controlled by the pressure change of the pneumatic channel 1000, it may be characterized in that it is directly adjusted by finger drive.

In the present invention, the first moving part may be characterized in that it includes a sample pretreatment and analysis function.

The second pneumatic valve 700 and the second moving part 800 control the movement of the microfluid passing through the operation chamber to the discharge part, and the second pneumatic valve is not directly connected to the pneumatic channel, It may be characterized in that the opening and closing is controlled by the polymer thin film 900 according to the pressure change of the microfluidic moving part by the operation of the first empty valve and the operation chamber.

In the present invention, the second moving part may be characterized in that it includes a sample pretreatment and analysis function.

In the present invention, the substrate is characterized in that the substrate is selected from the group consisting of glass, polyethylene terephthalate (PET), polymethyl methacrylate (PMMA), polycarbonate (PC), and polyethylene sulfone (PES). I can.

In the present invention, the polymer may be used as long as it is a polymer applied to a microfluidic device, but is preferably selected from the group consisting of rubber, linear low-density polyethylene (LLD-PE), and fluorodimethylsiloxane (PDMS). You can do it.

3 is a view for explaining the operation mechanism of the lab-on-a-chip system devised in the present invention, and the left panel shows a change that occurs when the pressure chamber 1100 is pressed with a finger. When pressure is applied by pressing the pressure chamber with a finger, the pressure is transmitted to the first pneumatic valve 400 and the operating chamber 600 through the pneumatic channel, thereby blocking the movement of the microfluid in the first moving part. 2 The pneumatic valve 700 is in an open state and discharges the microfluid in the operating chamber and the second moving part 800.

The right panel shows the changes that occur when the finger is swung. When the finger is released, the increased pressure in the pneumatic channel decreases, so that the first pneumatic channel and the operating chamber are restored to their original state, so that the microfluid corresponding to the volume of the operating chamber can move from the injection portion to the operating chamber through the first moving portion. . By repeating this, the movement of microfluids is controlled.

4 is a step-by-step view of the microfluidic control method.

First, in the initial state of the device according to the present invention, as shown in (1), since no pneumatic pressure is applied to the polymer thin film, the horizontal state is maintained. However, in order to operate the device, the polymer thin film must remain deformed toward the pneumatic channel due to the reduced pressure of the pneumatic channel as shown in (2). (2) To maintain the same state as (2), press the pressure chamber with your finger to drain the air from the pneumatic channel, and then close the pneumatic outlet with an acrylic block. After that, the sample is loaded and the finger movement of the pressure chamber is repeated to control the movement of the microfluid.

That is, step (2) is the basic state in which the device according to the present invention is operated, and from this time on, the first and second pneumatic valves, the operation chamber, and the first and second moving parts are connected to the pressure chamber and the pneumatic channel by the method of FIG. It is controlled by.

The lab-on-a-chip system according to the present invention is not affected by the depth, number of times, and finger driving characteristics at which the pressure chamber is pressed, and not only exhibits the same effect (Fig. 5), but also controls the amount of microfluids moving in the system. It can be easily adjusted by changing the diameter (Fig. 6).

In the present invention, it is characterized in that the first pneumatic valve 400 and the operating chamber 600 are connected to the pressure chamber 1100 by the pneumatic channel 1000 with the polymer thin film 900 interposed therebetween. I can.

In the present invention, the pressure of the pneumatic channel 1000 may be regulated by deformation of the pressure chamber 1100 due to external pressure.

In the present invention, the first pneumatic valve may be opened and closed by a polymer thin film 900 operated by the pneumatic channel 1000 connected to the pressure chamber 1100.

In the present invention, the operation chamber 600 may be compressed and depressurized by the polymer thin film 900 operated according to the pressure of the pneumatic channel 1000 connected to the pressure chamber 1100. .

In the present invention, the operation chamber 600 may be compressed and depressurized by a volume corresponding to the height of the first and second moving parts 500 and 800 and the pneumatic channel 1000.

In the present invention, the heights of the first and second moving parts and the pneumatic channel may be used without limitation as long as they can be engraved on the substrate, but may be preferably 0.001 to 1 mm.

In the present invention, the second pneumatic valve 700 is controlled according to the compression and decompression of the operation chamber 600 and the opening and closing of the first pneumatic valve 400, characterized in that the opening and closing according to the pressure of the operation chamber You can do it.

In another aspect of the present invention, (a) applying an external pressure to the pressure chamber of the lab-on-a-chip; (b) blocking the pneumatic discharge unit while external pressure is applied; (c) removing the external pressure applied in step (a); (d) injecting the sample into the injection unit; And (e) applying and removing external pressure to the pressure chamber; and a method for controlling microfluids of a lab-on-a-chip.

In the present invention, step (e) may be characterized in that it is repeated until the fluid is discharged to the discharge unit.

On the other hand, in the present invention, it was attempted to confirm whether the lab-on-a-chip system for multi-fluid analysis can be constructed by using the lab-on-a-chip system as one fluid driving unit.

That is, in another embodiment of the present invention, the lab-on-a-chip system is used as one fluid driving unit, and includes a first fluid driving unit for injecting a diluent and a second fluid driving unit for injecting a sample. The effect was confirmed by developing a microfluidic mixing device in which the system is controlled by one pressure chamber while being bundled together (FIG. 7).

In another embodiment of the present invention, a lab-on-a-chip system capable of detecting Escherichia coli using four fluid driving units independently controlled by two pressure chambers was developed (FIG. 8).

In another embodiment of the present invention, a lab-on-a-chip system for a blood cross-conformity test was developed using four fluid drive units controlled by one pressure chamber (FIG. 9).

In another embodiment of the present invention, a lab-on-a-chip system capable of extracting nucleic acids using four fluid drive units controlled by three pressure chambers was developed (FIG. 10).

Accordingly, in another aspect, the present invention includes the lab-on-a-chip as a fluid driving unit for driving one type of fluid, and a lab-on-a-chip using microfluidic control for multi-fluid analysis in which a plurality of fluid driving units are connected. It's about the chip.

In the present invention, the plurality of fluid drive units may be connected to have one discharge unit.

In the present invention, the plurality of fluid drive units may be connected to one pressure chamber.

In the present invention, the plurality of fluid driving units may have a plurality of pressure chambers.

In the present invention, the discharge unit connected to the plurality of fluid driving units may be connected to the injection unit of the fluid driving unit operated by another pressure chamber.

In the present invention, the fluid driving unit may be characterized in that it has a plurality of discharge units.

In the present invention, the plurality of fluid drive units are connected to a plurality of pneumatic valves operated by a plurality of pressure chambers to the second moving part, so that the fluid is moved to a specific discharge part through the adjustment of the pressure chamber. It can be characterized.

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

Example 1. Lab-on-a-chip system basic device performance check

1-1. Lab-on-a-chip system basic equipment manufacturing

Two layers of poly(dimethylsiloxane) having a thickness of about 3 mm and a PDMS film having a thickness of about 25 μm were inserted between them to fabricate a basic device for a lab-on-a-chip system. The mold of the PDMS layer was manufactured by photolithography, and SU-8 2005 (Microchem Corp., MA, USA) was coated on a silicon wafer with a thickness of 5 μm to prevent the structure of the pneumatic valve from sticking to the PDMS film. Then, SU-8 2100 was coated to obtain a thickness of 95 μm, and a PDMS precursor and a curing agent were mixed in a ratio of 10:1, and then injected into the SU-8 mold. After heating at 150° C. for 10 minutes, the PDMS layer was obtained by separating from the SU-8 mold. The PDMS film was obtained by spin-coating a mixture of a PDMS precursor and a curing agent in a ratio of 10:1 on a silicon wafer (1500 rpm for 60 seconds) and heating at 150° C. for 10 minutes.

The PDMS layer and the film were treated with oxygen plasma for 1 minute, and then incubated at 65° C. for 10 minutes to adhere to each other. First, a microfluidic moving part; Operating chamber; Pneumatic valve; And the PDMS layer containing the pressure chamber and the PDMS film on the silicon wafer were bonded, and then the assembly was removed from the silicon wafer, and this was combined with the PDMS layer containing the pneumatic channel.

1-2. Lab-on-a-chip system basic unit performance check

In order to confirm that the performance of the device manufactured in Example 1-1 is not affected by the characteristics of finger operation, as a result of measuring the volume of the fluid driven according to the depth at which the pressure chamber is pressed, as disclosed in FIG. 5A In the process of removing air from the pneumatic channel for the operation of the device, it was confirmed that a certain amount of fluid was driven by simply pressing deeper than the depth of the pressed pressure chamber.

In addition, it was confirmed that a certain amount of fluid was driven regardless of the time interval for pressing the pressure chamber (Fig. 5B), and when several people used the device, it was confirmed how it affected the performance of the device. It was confirmed that there was no significant difference in the amount of (Fig. 5C).

Since the amount of fluid that can be moved through the device corresponds to the volume of the operating chamber, it is expected that the amount of fluid that can be moved through the device can be adjusted according to the diameter of the operating chamber or the number of times the pressure chamber is pressed. As shown in graph 6A, it was confirmed that the amount of fluid moved varies depending on the diameter of the operating chamber and the number of times the pressure chamber is pressed, and the calculated volume of the operating chamber and the experimentally obtained value (drive It can be seen that the amount of fluid produced/device operation) is similar.

Example 2. Multi-fluid analysis device performance verification using lab-on-a-chip system

2-1. Sample mixing device fabrication and performance check

In the case of the device manufactured by the method of Example 1, by connecting a plurality of operating chambers and pneumatic valves for fluid drive to one pressure chamber, it is possible to simultaneously drive various samples through one pressure chamber and mix them at a desired ratio. have.

As a result of mixing the sample with the manufactured device, it was confirmed that the sample was diluted according to the diameter ratio of the operating chamber as disclosed in FIG. 7B. It was also confirmed that the concentration of the actual diluted sample almost coincides with the dilution concentration predicted through the diameter ratio of the operating chamber (FIG. 7C).

2-2. E. coli detection device fabrication and performance check

When E. Coli O157:H7, one of the representative food poisoning bacteria, is decomposed, an enzyme called β-galactosidase is released. When this enzyme reacts with an enzyme called CPRG, CPRG, which was originally yellow, turns red by the enzyme. It is converted into prominent CPR, and E. Coli O157:H7 can be detected (Fig. 8A).

In order to implement this by using the method of controlling microfluids by driving a finger according to the present invention, the apparatus disclosed in FIG. 8B was designed and manufactured by the method of Example 1.

First, by using a pressure chamber 1 E. Coli O157: and incubated for 10 minutes after the semi-mix H7 and Rai system buffer E. Coli O157: H7 by the decomposition, and then, by using CPRG a pressure chamber 2 and E. Coli O157: As a result of observing the color of the reaction chamber after mixing and reacting the H7-decomposed solution, it was confirmed that the color changed according to the concentration of E. coli (FIGS. 8C, D).

2-3. Blood cross-conformity test device fabrication and performance check

The cross-blood compatibility test is a test that finally confirms the side effects of hemolytic transfusion that may occur between the blood of the donor and the blood of the donor.It is confirmed by reacting the blood cells of the donor with the blood cells of the donor and the major test that checks by reacting the blood cells of the donor with the blood plasma of the donor. It consists of a minor test.

Existing blood cross-conformity tests require a complex process that requires skilled experts and expensive equipment such as centrifuges, so in the case of emergency patients who require blood transfusions as quickly as possible, cross-conformity tests are performed through conventional methods. The disadvantage is that it is difficult to perform. In addition, even when blood transfusion is required in a harsh environment, such as in developing countries, it is difficult to perform a blood cross-conformity test through the existing method.

As disclosed in Fig. 9A, a device capable of performing the experiment was designed by the method of the present invention. By incorporating an asymmetric membrane for plasma separation, the plasma of the donor and the recipient can be separated naturally. Subsequently, each separated plasma was allowed to cross-react with each blood using a pressure chamber.

9B is a photograph of the actually completed device. For the blood cross-conformity test, the blood of the donor and the donor must be separated into plasma and blood cells. To this end, a commercially available asymmetric membrane for plasma separation is partially compressed and applied to the device using a double-sided tape as shown in Fig. 9C. Internalized.

For the four possible possibilities of cross-conformity test by using the blood type combination of actual patients, the cross-conformity test (Off-chip) using the conventional method and the cross-conformity test (On-Chip) using the chip proposed in the present invention As a result of comparing the results, as shown in FIG. 10, when compared with the conventional method, a centrifuge was not used, no pipetting was required, and a small amount of blood (50 μL) was used to operate the finger within 10 minutes. You can quickly check the results of the cross-conformance test.

2-4. Production of nucleic acid extraction device

In order to perform nucleic acid extraction through a complicated process only by driving a finger, an apparatus as shown in Fig. 11A was designed. The nucleic acid extraction device consists of a total of three pressure chambers.

When the pressure chamber 1 is pressed, the pathogen is decomposed, the extracted internal nucleic acid is adsorbed on the surface of the glass bead in the glass bead chamber, and when the pressure chamber 2 is pressed, the washing buffer is adsorbed on the glass bead surface. It was designed to wash off impurities except for and press the pressure chamber 3 to elute the extracted nucleic acids to the desired discharge part.

As described above, specific parts of the present invention have been described in detail, and it will be apparent to those of ordinary skill in the art that these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereby. will be. Accordingly, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.

100: injection part
200: discharge unit
300: pneumatic outlet
400: first pneumatic valve
500: first moving part
600: operating chamber
700: second pneumatic valve
800: second moving part
900: polymer thin film
1000: pneumatic channel
1100: pressure chamber
1200: moving part

Claims (19)

A moving part 1200 through which the fluid moves; A pressure chamber 1100 driven by external pressure; A pneumatic channel 1000 through which compressed air moves; An injection unit 100 for injecting a fluid; A discharge unit 200 for discharging the fluid; And a pneumatic discharge unit 300;
The moving part and the pneumatic channel are separated by a polymer thin film 900,
The moving part includes first and second moving parts, and first and second pneumatic valves for controlling the movement of fluid; And an operating chamber for moving the fluid;
The first pneumatic valve and the operating chamber are connected to the pressure chamber by the pneumatic channel with the polymer thin film interposed therebetween,
The first pneumatic valve is opened and closed by a polymer thin film operated by a pneumatic channel connected to the pressure chamber,
The operating chamber is compressed and depressurized by a polymer thin film operated by a pneumatic channel connected to the pressure chamber,
The second pneumatic valve is a lab-on-a-chip using microfluidic control, characterized in that the second pneumatic valve is opened and closed according to the pressure of the moving part, which is adjusted according to compression and decompression of the operation chamber and opening and closing of the first pneumatic valve.
delete delete The lab-on-a-chip using microfluidic control according to claim 1, wherein the pressure of the pneumatic channel is controlled by deformation of the pressure chamber due to external pressure.
The lab-on-a-chip using microfluidic control according to claim 1, wherein the first moving part and the second moving part include sample pretreatment and analysis functions.
delete delete The lab-on-a-chip according to claim 1, wherein the operation chamber is compressed and depressurized by a volume corresponding to the height of the first and second moving parts and the pneumatic channel.
delete The lab-on-a-chip according to claim 1, wherein the polymer thin film is selected from the group consisting of rubber, linear low-density polyethylene (LLD-PE), and flodimethylsiloxane (PDMS).
The lab-on-a-chip according to claim 1, wherein the injection unit further includes a sample separation membrane.
(a) applying external pressure to the pressure chamber of the lab-on-a-chip according to any one of claims 1, 4, 5, 8, 10 and 11;
(b) blocking the pneumatic discharge unit while external pressure is applied;
(c) removing the external pressure applied in step (a);
(d) injecting the sample into the injection unit;
(e) applying and removing external pressure to the pressure chamber;
Lab-on-a-chip microfluidic control method comprising a.
The method of claim 12, wherein step (e) is repeated until a specific amount of fluid is discharged to the discharge unit.
Including the lab-on-a-chip according to any one of claims 1, 4, 5, 8, 10, and 11 as a fluid driving unit for driving one type of fluid, and a plurality of fluid driving units connected to each other for multi-fluid analysis Lab-on-a-chip using fluid control.
The microfluidic control for multi-fluid analysis according to claim 14, wherein the plurality of fluid driving units are connected to have one discharge unit and are connected to an injection unit of a fluid driving unit operated by another pressure chamber. Lab on a chip using.
15. The lab-on-a-chip according to claim 14, wherein the plurality of fluid driving units are connected to one pressure chamber.
The lab-on-a-chip according to claim 14, wherein the plurality of fluid drive units have a plurality of pressure chambers.
[15] The lab-on-a-chip according to claim 14, wherein the fluid driving unit has a plurality of discharge units.
The method of claim 14, wherein the plurality of fluid drive units are connected to a plurality of pneumatic valves operated by a plurality of pressure chambers to the second moving unit to move the fluid to a specific discharge unit through control of the pressure chamber. Lab-on-a-chip using microfluidic control for multi-fluid analysis, characterized in that there is.

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