KR102351481B1 - Micro platform for observing reaction of microfluids - Google Patents

Abstract

The present invention relates to a microplatform for observing microfluidic reactions, in which a microchannel 16 is formed long, a reaction chamber 15 is provided at a predetermined portion of the microchannel 16, and an injection hole 12 is formed. The stage 10 as a plate-shaped member, the elastic block 21 having a negative pressure channel 211 communicating with the microchannel 16 formed therein, and a roller bar generating negative pressure inside the microchannel 16 ( 22), and a membrane filter 13 installed at the inlet 12 to block the outside of the stage 10 and the inside of the microchannel 16, so that a very small amount of body fluid sample is transferred to the roller bar An object of the present invention is to provide a micro-platform characterized in that the microchannel 16 can pass due to the variability of (22), so that a high-resolution detection reaction test can be performed immediately even with a very small amount of sample.

Description

Micro platform for observing reaction of microfluids

The present invention relates to a platform for observing microfluidic reactions, and in particular, to a micro platform for observing microfluidic reactions configured so that the entire process of enzyme immunoassay (ELISA), which is a complex procedure, can be achieved in a small space with one device. .

A method for measuring antigen or antibody amount using an antigen-antibody reaction using an enzyme as a marker is generally referred to as an enzyme immunoassay. Enbvall, who developed this method, first used an immunosorbent and called it Enzyme-Linked ImmunoSorbent Assay (ELISA). ) as an Enzyme-LinKed ImmunoSpecific Assay (ELISA), the opinion prevails. ELISA is the most used experiment not only in the medical field but also in protein research because of the advantages of being able to analyze many samples at once as well as simple and accurate experimental method.

In the conventional ELISA, a 96-well plate in which 96 sample beakers or tubes are densely inserted vertically and horizontally into a plate with 96 insertion holes is used. do. And, since the amount of sample required should be enough to fill the beaker or tube to a certain level, especially in the case of expensive samples, a significant cost is required to perform the process. Therefore, when instantaneous on-site diagnosis is required, there is a problem that it is difficult to apply due to a problem that takes a long time and a problem of cost.

However, in the case of antibody immune response, it is widely used not only for academic experiments in research institutes but also for urgent prescriptions in a wide range of hospitals, so it needs to be carried out quickly for urgent treatment. However, since the conventional ELISA process takes considerable time, cost, and effort, it cannot meet the needs of on-site medical care in many cases, so that it is not possible to receive treatment through an accurate diagnosis in a timely manner.

As shown in FIG. 3, the reaction process consisting of ELISA includes the process of capturing the antibody, the process of binding the target antigen to the captured antibody, and the antibody-antigen conjugate again while binding the antibody in a sandwich form, and labeling the marker together with the antibody. It consists of a process of binding and a process of finally binding the enzyme substrate. In this case, it is necessary to inject a sample such as an antigen or antibody for each reaction step and wait for a long reaction, and a process of washing and removing unnecessary reactants is also required in the middle of each reaction. Therefore, an enormous amount of time is inevitably required for a single immune reaction, and when a conventional 96-well plate is used, a considerable amount of expensive samples is used so that the reaction can be visually determined, which incurs considerable costs.

On the other hand, in addition to the conventional 96-well plate, in the prior art, as a key tool for a simpler test, that is, a pregnancy diagnosis test through urine, sweat, or instant blood collection, or an instant test such as a blood sugar test, litmus paper made of pulp or similar Paper material is used. At this time, the litmus paper is coated with an antibody substance that changes color by reacting with the body fluid on a certain area. Inspection is performed by visually judging discoloration due to the reaction that occurs while reaching the

However, when capillary action is used as the principle of movement of body fluids in such litmus paper, there is a problem in that the reaction time cannot be accurately controlled.

This is because, in the case of blood, the viscosity varies according to age, so the reaction time by capillary pressure of litmus paper varies according to age. In addition, even in body fluids other than blood, the concentration of protein components in body fluids may vary according to age. There is a risk that the test results may appear significantly different due to differences in

Therefore, antibody immunoreaction analysis can be performed much faster than the 96-well plate used for conventional ELISA, and even a very small amount of sample can be analyzed, so the cost is much lower, and multiple reaction processes require intermediate washing. It is performed automatically, including the inclusion of the test, so the labor of the experimenter can be reduced, and multiple reactions are possible while being compactly made with a minimum size. A technology for a reaction device that can decisively contribute to prevention and prevention of diffusion is required.

In addition, the movement of body fluids, including blood, does not depend on the capillary phenomenon and moves completely by itself, so the phenomenon of blockage due to the protein that accumulates in the movement path can be fundamentally solved, but it can be used immediately, so non-specific reactions are excluded As a result, there is a demand for a technology for inspection equipment capable of deriving significantly more accurate results than inspection equipment using conventional litmus paper.

Patent Registration Publication No. 10-1515020 (Announcement Date: 2015. 04. 24)

Therefore, in the present invention, antibody immune response analysis can be performed much more quickly than the 96-well plate used in the conventional ELISA, and the analysis is possible even with a very small amount of sample, so the cost is much lower, and multiple reaction processes are possible. Since it is performed automatically, including intermediate washing, the labor of the experimenter can be reduced, and multiple reactions are possible while being made compact with a minimum size. , In addition, the movement of body fluids including blood does not depend on capillary action and moves completely by itself, so the phenomenon of blockage due to protein accumulating in the movement path can be fundamentally solved, but it can be used immediately, so that non-specific reactions can occur. It is intended to provide a micro-platform capable of deriving significantly more accurate results than the inspection equipment using conventional litmus paper by being excluded.

The micro platform according to the present invention for achieving this object includes a microchannel 16 through which a liquid can pass, a reaction chamber 15 formed in a chamber shape having a predetermined area in a predetermined portion of the microchannel 16, and a microchannel A stage 10 having a channel 16 and a reaction chamber 15 built therein, an injection hole 12 communicating with the outside is formed at the end of the microchannel 16, and a negative pressure channel communicating with the microchannel 16 Consists of an elastic block 21 having a 211 formed therein, and a roller bar 22 that generates negative pressure in the microchannel 16 by varying while pressing the elastic block 21 from the upper portion of the elastic block 21 . a micropump 20 and a membrane filter 13 installed in the inlet 12 to block the outside of the stage 10 and the inside of the microchannel 16; As a result, a very small amount of a bodily fluid sample can pass through the microchannel 16 due to the change of the roller bar 22 , so that even a very small amount of sample can be tested for high-resolution detection immediately.

Here, the membrane filter 13 is preferably coated with a sample reacting with the body fluid on or inside the membrane filter 13, and the sample body fluid passing through the membrane filter 13 reacts with the sample and then flows into the microchannel 16, The mixing for the reaction of the body fluid and the sample is facilitated by the negative pressure generated by the change of the roller bar 22 .

Also, the elastic block 21 is preferably disposed on the stage 10 so that the negative pressure channel 211 is parallel to the microchannel 16, and the end opposite to the injection hole 12 in the microchannel 16 By forming a connection channel 212 connecting the end of the negative pressure channel 211 and the negative pressure channel 211, the change in the sound pressure inside the negative pressure channel 211 according to the change of the roller bar 22 is transmitted to the microchannel 16 and the microchannel (16) Sound pressure is generated and changed inside.

In addition, the reaction chamber 15 preferably has two channel communication ports 154 communicating with the microchannel 16 on one side and the microchannel 16 on the other side. At the center of the reaction chamber 15, a reaction zone 152 in the form of a two-dimensional figure in which the long side direction is disposed in the longitudinal direction of the microchannel 16 is formed in a constant area, and the channel communication The sphere 154 is connected to the bottom of the reaction zone 152, and each channel communication hole 154 is on one side and the other side of the reaction zone 152 in an imaginary center line mountain passing in the longitudinal direction of the reaction zone 152. each is formed

In this case, a step surface 153 that is preferably higher than the reaction zone 152 and lower than the upper surface of the stage 10 is formed between the reaction zone 152 and the upper surface of the stage 10, so that the reaction zone 152 is formed. Even if the passing sample overflows in the reaction zone 152 , it can be accommodated in the reaction chamber 15 .

At this time, a transparent cap 151 in the form of a transparent circular plate larger than the area of the reaction zone 152 and the stepped surface 153 is preferably installed on the upper portion of the reaction chamber 15, and the transparent cap 151 is a transparent cap. (151) A double-sided tape or an adhesive layer is provided and fixed to the edge portion in contact with the upper surface of the stage 10 from the bottom surface.

Also preferably as a vertical column-shaped member having a constant height, the center of the bottom surface protrudes and is in close contact with the entire reaction zone 152 , the bottom edge is in close contact with the upper surface of the stage 10 as a plane, and the center of the top surface is the reaction zone It is formed with the upper reaction zone 1552 having the same shape as 152 , and the upper edge is formed with an upper stepped surface 1553 having the same shape as the stepped surface 153 of the reaction chamber 15 , and has a bottom surface from an upper surface inside. A reaction chamber floating column 155 is provided in which two microchannels 1551 of a penetrating form are formed and configured.

At this time, the upper portion of the reaction chamber floating column 155 may be preferably watertightly inserted into the beaker 156 constituting a 96-well plate for a conventional immune response test.

And each of the two vertical channels preferably protrudes more than the bottom surface of the reaction chamber floating column 155 so that the end can be inserted into the communication hole formed in the reaction chamber 15 .

The microplatform according to the present invention can perform antibody immune response analysis much faster than the 96-well plate used for conventional ELISA performance, and it is possible to analyze even a very small amount of sample, so the cost is much lower, and multiple The reaction process is automatically performed including intermediate washing, reducing the labor of the experimenter, and since it is made with a minimum size and compact, multiple reactions are possible. Also, the movement of body fluids, including blood, does not depend on capillary action and moves completely by itself, so the blockage of the movement path due to the protein accumulating can be fundamentally solved, but it can be used immediately, so it is non-specific By excluding the negative reaction, there is an effect of being able to derive significantly more accurate results than the inspection equipment in which the conventional litmus paper is used.

1 is a conceptual diagram of each component constituting the antigen-antibody reaction and the reaction process;
2 is a perspective view of a micro platform according to an embodiment of the present invention;
3 is a plan view of FIG. 2;
4a and 4b are side cross-sectional views showing the absorption process of blood at the inlet;
5A and 5B are side cross-sectional views showing the principle of the micropump;
6 is a cross-sectional side view of the reaction chamber;
Figure 7a is a perspective view of the embodiment to which the reaction chamber flotation column is applied in Figure 2;
7b is an exploded side cross-sectional view of the reaction chamber flotation column and reaction chamber;
7c is a partial enlarged view of an exploded side cross-sectional view of the reaction chamber flotation column and the reaction chamber;
8 is a conceptual view to which the reaction chamber flotation column is applied to FIGS. 4A to 5B;
9 is a measurement accuracy graph of a conventional diagnostic kit and a test platform according to the present invention;

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

The micro platform according to the present invention is composed of a plate-shaped stage 10 having a microchannel 16 formed therein, and a membrane filter 13 .

As shown in FIGS. 2 and 3 , the stage 10 is a reaction chamber in which a microchannel 16 through which a liquid can pass is formed long, and a predetermined portion of the microchannel 16 is formed in an expanded form. (15) is provided, is formed at the end of the microchannel 16 is a plate-shaped member in which the injection hole 12 for communicating the outside and the microchannel 16 with the outside is formed.

More specifically, the plate-shaped member constituting the stage 10 may be manufactured in the form of one joint body 19 in which two plates, the lower body 191 and the upper body 192, are joined to each other.

At this time, the microchannel 16 and the reaction chamber 15 may be manufactured by being engraved in the form of a pattern on the bottom surface of the upper body 192 or engraved on the top surface (not shown) of the lower body 191 as shown in FIG. 3A . have. In this case, for bonding between the upper body 192 and the lower body 191, any known bonding method such as adhesive, tape, or thermal fusion may be adopted.

In this way, when the upper body 192 and the lower body 191 are bonded to each other, and the microchannel 16 is formed in either one of the pattern-shaped processing, it does not go through a difficult process of processing in the form of a perforation inside one plate body. Even if it is not, the manufacturing of the stage 10 may be made through a simple process.

Alternatively, the microchannel 16 is disposed between the lower body 191 and the upper body 192, according to the embodiment shown in FIGS. 3B to 5B, to bond the lower body 191 and the upper body 192 to each other. It may be formed in the form of a pattern on the adhesive panel 193, which is a plate-shaped member including an adhesive component. In this case, the microchannel 16 formed in a pattern shape may be engraved in a shape penetrating the adhesive panel 193 . The adhesive panel 193 will be described later in more detail.

The membrane filter 13 is installed in the inlet 12 , and acts to block the outside of the stage 10 and the inside of the microchannel 16 . However, blocking at this time means blocking in a state where the fluid can pass.

The membrane filter 13 may be installed in the form of bonding to the upper surface of the stage 10 with the filter adhesive layer 131 provided on the lower surface of the membrane filter 13 as shown in FIGS. 3B and 4A and 4B . have. Here, the filter adhesive layer 131 may be a conventional adhesive or a double-sided tape.

In particular, the membrane filter 13 is coated with a sample reacting with the body fluid on or inside the membrane filter 13 , and the sample body fluid passing through the membrane filter 13 reacts with the sample and then flows into the microchannel 16 , Mixing for reaction is facilitated by the negative pressure generated in the variable negative pressure channel 211 of the roller bar 22 . That is, the driving force for allowing the sample body fluid to pass through the membrane filter 13 is provided from the micropump 20 .

Alternatively, as shown in FIG. 3b , a pad through which a liquid can pass between the membrane filter 13 and the inlet 12 is a secondary antibody that recognizes and binds the primary antibody bound to the antigen contained in the body fluid S1. A secondary antibody pad 14 coated with (S2) is disposed, the secondary antibody pad 14 has a smaller area than the membrane filter 13, and a membrane filter ( By providing an adhesive layer that adheres 13) to the upper surface of the stage 10, the body fluid (S1) passing through the membrane filter 13 passes through the secondary antibody pad 14, and the body fluid (S1) and the secondary antibody The secondary antibody-antigen complex (S3) formed by the reaction of (S2) may enter the microchannel 16 .

In addition, a micropump 20 may be installed so that the sample can be moved forward or backward through the inside of the microchannel 16 .

The micropump generates negative pressure inside the microchannel 16, so that the bodily fluid S1 passes through the membrane 13 filter and is sucked into the microchannel 16, or the body fluid S1 inside the microchannel 16 move the

The micropump 20 is a microchannel by varying the elastic block 21 having a negative pressure channel 211 communicating with the microchannel 16 formed therein, and pressing the elastic block 21 from the upper portion of the elastic block 21 . (16) It consists of a roller bar 22 that generates a negative pressure inside and is manually driven, or it is composed of a syringe pump (not shown) and can be automatically driven.

When the micropump is implemented manually, the micropump 20 is an elastic block 21 in which a negative pressure channel 211 communicating with the microchannel 16 is formed therein, as shown in FIGS. 5a and 5b; It consists of a roller bar 22 that generates a negative pressure inside the microchannel 16 by being variable while pressing the elastic block 21 from the top of the elastic block 21 .

The elastic block 21 is made of an elastic material, and when the upper portion of the negative pressure channel 211 is pressed, elastic deformation is transmitted to the negative pressure channel 211 so that the pressed portion is blocked. If the elastic block 21 is pressed in place, only the pressed portion is blocked, but if the elastic block 21 is pushed forward or backward while pressing the elastic block 21 with the roller bar 22 as in the present invention, FIGS. 5A and 5B are time sequence As a result, the clogged portion of the negative pressure channel 211 is moved, so that forced airflow movement within the microchannel 16 communicating with the negative pressure channel 211 occurs. That is, since the region where the negative pressure is formed is moved, if there is a liquid marked in black as shown in FIG. 5A inside the microchannel 16, it is moved in the direction shown in FIG. 5B.

And the elastic block 21 is composed of a lower plate 21b and a bumper 21a as shown in FIGS. 2 and 3, so that the negative pressure channel 211 on the bottom surface of the bumper 21a is processed in the form of a pattern and then the bumper ( 21a) and the lower plate 21b may be joined to each other.

The micro-platform according to the present invention communicates with the micro-channel 16 so that the micro-pump 20 in which the negative pressure inducing point is movable, not in a static state, communicates with the micro-channel 16. (16) Since it can be moved inside, the consumption of expensive samples, which is a problem in immune reaction experiments such as ELISA, which previously required a large amount of sample, can be significantly saved, and the reaction experiment can be performed even with a very small amount of body fluid. The time required for the experiment is significantly reduced, so it can be used even in situations where an urgent experiment is required in the field.

In addition, the problem of low accuracy of the test due to the difference in viscosity according to age, constitution, and other conditions, which was a problem in test kits that moved body fluids using a micro-capillary structure made of pulp, such as a pregnancy or diabetes test using a conventional litmus test paper, is fundamentally a problem. has the effect of being resolved.

This is because, as described in the background section above, when the viscosity of the protein contained in the body fluid is different, the protein component accumulates in the capillary while the body fluid moves through the microstructure of the pulp material according to the capillary phenomenon and blocks the capillary. This is because body fluid does not reach the original state until the antibody applied to the point spaced apart by a certain interval to react with the antigen.

In addition, in the test kit using the conventional litmus test paper, the movement speed is fixed at the rate of propagation by the capillary phenomenon because the movement of body fluids such as blood, sweat, or urine depends on the micro-capillary action of the pulp material constituting the litmus test paper. There is a problem in that it is impossible to control the movement speed of the body fluid depending on the situation.

On the other hand, in the microplatform according to the present invention, since the movement of body fluid is driven by the micropump 20 that variably moves the negative pressure generating point, the movement speed of the body fluid can be freely controlled, so that the body fluid can be stopped or The rapid movement can be freely adjusted.

Meanwhile, the elastic block 21 is disposed on the stage 10 as shown in FIGS. 5A and 5B so that the negative pressure channel 211 is parallel to the microchannel 16, and the injection hole in the microchannel 16 is A connection channel 212 connecting the opposite end of the 12 and the end of the negative pressure channel 211 is formed, so that the change in the sound pressure inside the negative pressure channel 211 according to the change of the roller bar 22 is reduced to the microchannel ( 16) to generate a negative pressure inside the microchannel 16.

That is, the sound pressure channel 211 and the microchannel 16 inside the elastic block 21 may be connected in a line in parallel, but they are arranged to be reversed as shown in FIGS. 5A and 5B, as if the negative pressure channel 211 and the microchannel (16) is formed in a bent shape, and by providing a connection channel 212 to connect each other, the volume occupied by the micro platform can be minimized, so that it is extremely easy to carry and install. That is, it has the same effect as being able to carry long-made equipment by folding it in half.

In particular, when more than one stage of immune response test is required, in the prior art, there was no equipment that could be performed immediately on site, so it was a reality that the urgent needs of patients could not be met. The reaction test can be made on the spot and precisely on the fly, and a compact arrangement between the microchannel 16 and the negative pressure channel 211 is made so that the portability of equipment, which is particularly important for this purpose, can be greatly improved.

The reaction chamber 15 may have two embodiments: an embodiment in which the reaction chamber 15 is formed on the upper surface of the upper body 192 , and an embodiment in which the reaction chamber 15 is formed between the upper body 192 and the lower body 191 .

In the embodiment in which the reaction chamber 15 is formed between the upper body 192 and the lower body 191, as shown in FIG. 3A, the microchannel 16 is expanded in some section to an area that allows observation of the reaction. form is formed. At this time, as shown in FIG. 3A, the capture antibody (S4) is pre-coated on the area formed by the reaction chamber, and the body fluid (S1) passes through the secondary antibody pad to form the secondary antibody-antigen complex (S3) in the reaction chamber. Upon reaching (15) and reacting with the capture antibody (S4), a secondary antibody-capture antibody-antigen complex (S5) is formed inside the reaction chamber (15). In this case, the upper body 19 at a position corresponding to the upper portion of the reaction chamber 15 is made transparent so that the reaction process can be observed.

In an embodiment in which the reaction chamber 15 is formed on the upper body 192, as shown in FIG. 3b, the reaction chamber 15 includes a cut-off section BS formed in a predetermined portion of the microchannel 16, and a cut-off section Channel communication hole 154, which is two through-holes formed in the upper body 192 to correspond to the positions of the two opposite ends of the two micro-channels 16 separated by (BS), and two channel communication holes as a transparent plate An adhesive layer 1511 for installing a cap that is disposed to cover the transparent cap 151 and the two channel communication holes 154 at the same time to bond the upper surface of the transparent cap 151 and the upper body 192 to each other. ) is made of

At this time, the space surrounded by the bottom surface of the transparent cap 151 and the top surface of the upper body 192 and the inside of the adhesive layer 1511 for installing the cap is formed as a reaction zone 152 in which a reaction occurs.

At this time, in the embodiment of FIG. 3B as in the embodiment of FIG. 3A , the capture antibody S4 is pre-coated on the area formed by the reaction chamber, so that the same reaction as in the embodiment of FIG. 3A can be performed.

For reference, the symbols shown in Fig. 1 each indicate S1 for a bodily fluid sample containing antigen, S2 for secondary antibody (S2), S3 for secondary antibody-antigen complex (S3), S4 for capture antibody (S4), and S5 for The secondary antibody-capturing antibody-antigen complex (S5) and S6 will be referred to as identification enzymes.

And, although not shown, a washing solution (not shown) may be introduced later through the inlet 12 . At this time, when the washing solution passes through the reaction zone 152, non-specific free antigens (not shown) that are not bound to the immobilized capture antibody (S4) during the passage of the washing solution are removed. After that, when the substrate (S6) is injected through the injection port 12, the substrate (S6) reacts with the secondary antibody-capturing antibody-antigen complex (S5) immobilized in the reaction zone 152 as shown in FIG. 1 . While the secondary antibody-capturing antibody-antigen complex (S5) has a specific color or is displayed with a fluorescent color, the presence and concentration of the antigen can be identified.

Such a complex ELISA reaction test can also be performed with the microplatform according to the present invention, and a simpler immunoassay method than ELISA can be performed immediately and in a short time.

Also, for example, when a microplatform is used as a simple pregnancy test, a pregnancy test based on a conventional litmus test strip reacts with HCG recognized as an antigen and has a colored antibody attached to the target point, but Many errors can occur due to differences in concentration for each constitution, and in particular, even if an abnormality occurs in the pregnancy test, it is difficult to identify with the naked eye.

On the other hand, in the microplatform according to the present invention, the reaction zone 152 can be observed with the naked eye, and the body fluid sample is immediately transferred to the reaction zone 152 without clogging the body fluid sample even if the age or constitution is different, so it depends on the capillary phenomenon of the pulp material. Accuracy and reliability can be significantly improved compared to one transfer method.

In addition, a transparent cap 151 in the form of a transparent circular plate is installed above the reaction chamber 15 for visual observation of the reaction zone 152 .

In this case, the transparent cap 151 may be made of polycarbonate of a transparent material, but is not necessarily limited thereto as long as it is a transparent material. The fixing of the transparent cap 151 is between the edge of the bottom of the reaction cap and the stage 10, that is, at the edge among the points where the transparent cap 151 is installed among the upper surface of the upper body 192, an adhesive layer for cap installation such as double-sided tape ( 1511) is formed. Therefore, even if deterioration occurs, such as a decrease in transparency of the reaction cap, it can be easily replaced with a new one.

On the other hand, the reaction chamber floating column 155 may be installed so that the position of the reaction chamber 15 is raised and the length of the microchannel 16 is further extended to more easily control the movement of the sample body fluid.

The reaction chamber floating column 155 is a member of a vertical column shape having a constant height as shown in FIGS. 7A to 7C , and the bottom is in close contact with the upper surface of the stage 10, that is, the upper body 192, and the center of the upper surface is An upper reaction zone 1552 having the same shape as the reaction zone 152 is formed, and two vertical microchannels 1551 penetrating from the top to the bottom are formed therein. And the installation of the reaction chamber floating column 155 may be made by adhesion by the column adhesive layer 1555 as shown in FIG. 7B .

Here, the two vertical microchannels 1551 are disposed at corresponding positions to be connected one by one to the two channel communication holes 154 formed in the upper body 192, respectively.

A cover 156 is coupled to the upper portion of the reaction chamber flotation column 155 . Referring to FIGS. 7B and 7C , since the reaction is performed under the ceiling of the cover 156 , the upper surface of the cover 156 or the entire cover 156 is made to be transparent.

In particular, the upper portion of the reaction chamber floatation column 155 may be tightly inserted into the beaker constituting a 96-well plate for a conventional immune response test as shown in FIGS. 7A and 7B . That is, the beaker constituting the 96-well plate may be used as the cover 156 .

In this case, before the cover 156 or the 96-well beaker is installed on the upper part of the reaction chamber flotation column 155, the capture antibody (S4) or the secondary antibody (S2) is placed on the ceiling of the cover 156 or the ceiling of the 96-well beaker. It can be applied in advance.

The reaction chamber flotation column 155 is configured in this way to perform three functions.

First, by floating the reaction chamber 15 itself much higher than the upper surface of the stage 10, it makes it easier to observe the reaction process.

Second, the length of the microchannel 16 has the effect of extending as much as twice the height of the reaction chamber floating column 155, so that the control of the sample passing through the microchannel 16 becomes easier. This is because, in particular, in order to pass two or more samples into the microchannel 16 for a sequential reaction, the more the distance between them is secured, the easier it is to control.

Third, in the conventional immune response test using a 96-well plate, the method of pre-immobilizing the aforementioned capture antibody (S4) in the beaker 156 constituting each 96 well may be the same as in the prior art, but in the prior art, each Since the color change was observed while observing the beaker 156 from the side by administering various reactants to the beaker 156 itself, considerable expensive reagents are inevitably consumed. As a result, it is almost impossible to immediately conduct an urgent immune response test in the field with 96 wells.

In the microplatform according to the present invention, each 96-well beaker (S4) to which the capture antibody (S4) is immobilized so that it can be recycled as it is rather than discarding the conventional 96-well plate while overcoming the disadvantages of the 96-well plate used in the conventional ELISA reaction 156) is used as it is, and it is the reaction chamber floating column 155 that can be turned over and combined.

Here, two vertical microchannels 1551 connected from the bottom to the top of the reaction chamber floating column 155 are built-in, one serves as a passage for supplying the sample to the upper reaction zone 1552, and the other serves as a passage for the upper reaction of the sample. It serves as a passage for exhaust from the zone 1552 .

Even when the reaction chamber floating column 155 is installed, the overall procedure of the inspection is almost the same as that in the case where only the reaction chamber 15 is formed. However, as shown in FIG. 8, the distance at which the secondary antibody-antigen complex (S3) reaches the upper reaction zone 1552 and the secondary antibody-capturing antibody-antigen complex (S5) are discharged from the reaction zone 152 Since the distance becomes longer, the moving distance of the roller bar 22 may be slightly longer.

For reference, FIG. 8 shows that body fluid (S1) from the outside passes through the secondary antibody pad 14 and enters the injection port, binds to the secondary antibody (S2) to form a secondary antibody-antigen complex (S3), and then microchannel After proceeding along (16), it climbs to the upper part of the reaction chamber flotation column 155 on the vertical microchannel 1551 through the channel duct 154, and binds to the capture antibody (S4) to thereby bind the secondary antibody-capturing antibody-antigen. The entire process of the ELISA forming the complex (S5) is shown in sequence.

Meanwhile, FIG. 9 is a graph related to the accuracy of measurement results when the microplatform according to the present invention is used and when a test kit in the form of a conventional litmus test strip is used. For reference, the expression 'Proposed LOC (Lab-On-a-Chip)' in FIG. 9 refers to the micro platform according to the present invention, and the expression 'Lateral flow assay' refers to a test kit using a conventional litmus test paper. refers to

In FIG. 9 , a case in which an error due to interference occurs with respect to different concentrations of albumin containing troponin I and beta antigen is exemplified. As shown in FIG. 9 , it can be seen that the error rate due to interference is much higher in the case of the conventional test kit compared to the microplatform according to the present invention.

The present invention described above is not limited by the above-described embodiments and the accompanying drawings, and it is common in the technical field to which the present invention pertains that various substitutions, modifications and changes can be made within the scope without departing from the technical spirit of the present invention. It will be clear to those who have the knowledge of

S1: body fluid S2: secondary antibody
S3: secondary antibody-antigen complex S4: capture antibody
S5: secondary antibody-capturing antibody-antigen complex S6: substrate
10: stage 12: injection hole
13: membrane filter 14: secondary antibody pad
15: reaction chamber 16: microchannel
19: bonding body 20: micropump
21: elastic block 22: roller bar
131 filter adhesive layer 151 transparent cap
152: reaction zone 153: adhesive side
154: channel communication port 155: reaction chamber floating column
156: cover 191: lower body
192: upper body 193: adhesive sheet
211: sound pressure channel 212: connection channel
1511: adhesive layer for cap installation 1551: vertical microchannel
1552: upper reaction zone 1555: column adhesive layer

Claims (8)

A microchannel through which a liquid can pass is formed therein, a reaction chamber in which an antibody is fixed is provided so that an immune reaction occurs in a predetermined portion of the microchannel, and is formed at an end of the microchannel to communicate the microchannel with the outside. A plate-shaped stage with an injection hole formed therein;
a membrane filter installed at the inlet to block the microchannel from the outside;
A secondary antibody pad disposed between the membrane filter and the inlet, allowing a liquid to pass through, and coated with a secondary antibody that binds to an antigen contained in body fluid.
Micro-platform, characterized in that it comprises.
According to claim 1,
A micropump is further provided for generating a negative pressure inside the microchannel, allowing the bodily fluid to pass through the membrane filter and sucking it into the microchannel, or for moving the bodily fluid inside the microchannel,
The micropump is composed of an elastic block having a negative pressure channel communicating with the microchannel formed therein, and a roller bar that generates negative pressure in the microchannel by varying while pressing the elastic block from an upper portion of the elastic block. Micro-platform, characterized in that it is driven or is automatically driven by a syringe pump.
3. The method of claim 2,
The elastic block is disposed on the upper part of the stage so that the sound pressure channel is parallel to the microchannel, and a connecting channel connecting the end of the microchannel and the end of the sound pressure channel is formed, thereby forming the microchannel and the connecting channel and a passage through which the negative pressure channel and the negative pressure channel are connected in parallel are formed in such a way that they are bent around the connection channel.
According to claim 1,
The secondary antibody pad has a smaller area than the membrane filter, and an adhesive layer for bonding the membrane filter to the upper surface of the stage is installed on the edge of the secondary antibody pad, so that the total amount of body fluid passing through the membrane filter is The microplatform, characterized in that passing through the secondary antibody pad, the secondary antibody-antigen complex formed by the reaction of the body fluid with the secondary antibody enters the microchannel.
According to claim 1,
The stage is formed by surface bonding of a lower body and an upper body in the form of a thin plate,
An adhesive panel is inserted between the lower body and the upper body to bond the lower body and the upper body,
The microchannel is formed in the form of a pattern on the adhesive panel,
The injection hole is a through hole formed in the upper body, and is formed in a vertical upper position of one end of the microchannel.
6. The method of claim 5,
a disconnection section formed in a predetermined portion of the microchannel;
A channel communication hole, which is two through-holes formed in the upper body, corresponding to the two end positions of the two microchannels separated by the cut-off section, and a transparent plate, a transparent plate disposed to cover the two channel communication holes at the same time A reaction chamber is provided that includes a cap and an adhesive layer for installing a cap disposed to surround both the channel communication ports and bonding the transparent cap and the upper surface of the upper body,
A space surrounded by the bottom surface of the transparent cap, the top surface of the upper body, and the inside of the adhesive layer for installing the cap is a reaction zone in which a reaction occurs.
6. The method of claim 5,
A disconnection section is formed in a predetermined portion of the microchannel,
Two through-hole channel communication holes are formed in the upper body to correspond to the two end positions of the two microchannels separated by the cut section facing each other,
As a vertical column-shaped member having a constant height, a vertical microchannel is formed in which two through-holes are drilled in parallel in the vertical longitudinal direction of the member at the same interval as the two channel communication holes, and a transparent material cover is formed on the upper part There is provided a reaction chamber flotation column consisting of a combination,
The microplatform, characterized in that the reaction chamber floating column is arranged so that the two channel communication port and the two vertical microchannels are connected.
8. The method of claim 7,
The cover is a cup-shaped member made of a transparent resin material or a microplatform, characterized in that it is a beaker container that is a component of a conventional 96-well plate.

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