KR101033206B1 - Microfluidic Channel and Microfluidic Device for Capturing Target, and Microfluidic Analyzing System Therewith - Google Patents

Abstract

The present invention relates to microfluidic channels and microfluidic devices for target capture and microfluidic analysis systems comprising the same. The present invention, the main channel; A target capture part formed in a part of the main channel and provided with a fluid discharge hole; A membrane which opens and closes the fluid discharge hole by being attached to and detached from a lower surface of the target capture unit; And a microfluidic channel for target capture including a drainage channel disposed between the fluid discharge hole and the membrane to discharge the fluid passing through the fluid discharge hole, and a microfluidic device and a microfluid analysis system including the same. do.

Microfluidic Devices, Targets, Sensitives, Capture, Antigens, Antibodies

Description

Microfluidic Channel and Microfluidic Device for Capturing Target, and Microfluidic Analyzing System Therewith}

The present invention relates to a microfluidic channel for capturing a target, a microfluidic device, and a microfluidic analysis system including the same, and can simultaneously capture and analyze various types of targets, thereby dramatically reducing analysis time and accuracy of analysis. And it relates to a microfluidic channel and a microfluidic device and a microfluidic analysis system including the same for the target capture to improve the reliability of the analysis results with excellent reproducibility.

Chemical disease analysis methods are indispensable for the physiological study of the disease, and are particularly essential for identifying the disease and identifying the patient's condition. Among these, the blood test is known as the most basic and important method of the test for determining the disease of the patient. For example, concentrations of various proteins, such as cytokines and complements in plasma, are known to be directly related to disease. In addition, it is possible to distinguish cancer types by detecting biomarkers of specific tumors from the blood, and detect cardiac makers such as C-reactive protein (CRP) and myglobin. By doing so, it is possible to make a fast and stable diagnosis of coronary heart disease. These blood tests are based on measuring the concentration of markers present in biological samples using specific binding reactions between ligand-receptors or antigen-antibodies.

Meanwhile, as microfluidic dynamics have recently been applied to various biological assays, marker detection methods based on microfluidic dynamics, which are more efficient and sensitive than conventional marker detection methods, have been proposed.

Microfluidic dynamics can be used to reduce the amount of expensive samples such as reagents and biochemicals used in the analysis due to the reduction of the system, and can be analyzed at a relatively lower cost than conventional methods due to the ease of automation.

In this paper, a paper published in 2002, "Method for Detection of Particle-Based Markers Using Magnetic Forces (choi et al, Lab Chip, 2002)", captured magnetic particles with a plate-shaped electromagnet formed by a microprocess and produced particles-antigen-antibody- The present invention discloses a technique of measuring the concentration of a marker by reacting with a secondary antibody in order and removing the particles by removing the magnetism of the electromagnet when the measurement is completed. This method is capable of repeated measurements, but the manufacturing process for precisely aligning the positions of the electromagnets and the electrodes, which are considered to affect the performance of the system, is very difficult. In addition, if the flow rate is increased to reduce the measurement time, particles that are not trapped in the electromagnet may fall out of the shear stress of the fluid, and there is a problem that it is difficult to control the number of particles that are trapped.

In addition, in the 2001 paper "Sato et al, Anal. Chem. 2001", the method of capturing particles using the dam structure, the antigen and the antibody combined with the fluorescent substance were captured. Are sequentially injected to have a structure of particle-antigen-antibody-secondary antibody and to determine the concentration of antigen by measuring the intensity of fluorescence generated under a microscope. This method has shortened the incubation time and reduced the total analysis time, and the principle of operation is the same as the general measurement method (antigen-antibody reaction), but since it is difficult to permanently form dams that trap particles, It cannot be captured and the reusability of the device is impossible, making it difficult to repeat the measurement. In addition, since a non-constant number of particles are measured in a stacked form, an error may occur due to a change in fluorescence intensity according to the particle number difference, and the reproducibility of the measurement may be low.

In addition, the article published in 2007, "Method for Detection of Particle-Based Markers Using the Characteristics of Microfluidics (Yang et al, Lab Chip, 2007)," introduced the concept of "particle cross over", enabling continuous measurement and changing antigens. A technique for continuously measuring the concentration of is disclosed. However, since this method measures the intensity of fluorescence while the particles are not captured and flows, sensitivity changes are expected depending on the flow rate and the length of the channel in which the culture is performed, and there is a problem that an error occurs depending on the position of particles in the channel.

As such, there have been many studies on microfluidic detection devices based on microfluidics, but it is possible to inspect specific markers in the blood at high sensitivity and high speed, and to capture particles in a single layer, thereby reducing measurement errors. The developed system has not been developed so far, and continuous research is required.

The present invention for solving the above problems, it is possible to capture and analyze a variety of targets at the same time can significantly reduce the analysis time, excellent accuracy and reproducibility of the analysis can be obtained a highly reliable analysis results It is to provide a microfluidic channel and a microfluidic device for target capture and a microfluidic analysis system including the same.

The present invention to achieve the above object, the main channel; A target capture part formed in a part of the main channel and provided with a fluid discharge hole; A membrane which opens and closes the fluid discharge hole by being attached to and detached from a lower surface of the target capture unit; And a drain channel disposed between the fluid discharge hole and the membrane to discharge the fluid passing through the fluid discharge hole.

Preferably, the fluid discharge hole is such that the size of the upper portion is larger than the size of the lower portion.

Preferably, the membrane is operatively controlled by either pneumatic or piezoelectric elements.

Meanwhile, a target detection unit for detecting a target captured by the target capture unit is provided above the target capture unit. Preferably, the target detection unit is composed of a double cladding optical fiber, the excitation light is transmitted to the core portion of the double cladding optical fiber, and the generated optical signal is transmitted to the first cladding portion surrounding the outer side of the core portion .

On the other hand, according to another embodiment of the present invention, the microfluidic channel device for target capture, the target storage unit for storing a target; A first sensing material storage unit storing a first sensing material primarily coupled to the target; A second sensing material storage unit storing a second sensing material which is secondarily coupled to the target; A flow controller for controlling the flow of the fluid stored in the target storage unit, the first sensing material storage unit, and the second sensing material storage unit; And a microfluidic channel device for target capture comprising a microfluidic channel as described above.

In addition, according to another embodiment of the present invention, the present invention, a microfluidic analysis system for target analysis, the target storage unit for storing a target; A first sensing material storage unit storing a first sensing material primarily coupled to the target; A second sensing material storage unit storing a second sensing material which is secondarily coupled to the target; A flow controller for controlling the flow of the fluid stored in the target storage unit, the first sensing material storage unit, and the second sensing material storage unit; Microfluidic channels as described above; A fluid flow controller for controlling the operation of the flow controller and the membrane; And a target detection signal acquisition unit configured to acquire an optical signal according to the target detection.

The microfluidic device for target capture of the present invention and the microfluidic analysis system including the same can simultaneously capture and analyze various types of targets, thereby significantly reducing the analysis time.

In addition, according to the present invention, it is possible to capture a target to be analyzed as a single layer, there is an advantage of reducing the measurement error due to non-uniform lamination and high accuracy and reproducibility of analysis.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. First of all, in adding reference numerals to the components of each drawing, it should be noted that the same reference numerals are used to refer to the same components even though they are shown in different drawings. In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

In addition, the following will describe a preferred embodiment of the present invention, but the technical idea of the present invention is not limited thereto and may be variously modified and modified by those skilled in the art.

Unless stated otherwise in the present specification, "target" refers to an object to be captured, detected, or analyzed, and specific examples thereof include antigens, DNA, proteins, and the like.

In addition, in the present specification, a "sensing substance" is a substance that binds to or reacts with a target, and specific examples thereof include an antibody and the like.

1 is a perspective view of a microfluidic device for target capture according to a preferred embodiment of the present invention, and FIG. 2 is a schematic diagram of a microfluidic analysis system including a microfluidic device for target capture according to a preferred embodiment of the present invention. .

The microfluidic device 10 for capturing a target according to an exemplary embodiment of the present invention includes a base 12, a plurality of fluid reservoirs installed in the base 12, and a plurality of flows for controlling the flow of the fluid. Controller, and microfluidic channel 40 for target capture.

The fluid reservoir includes a target reservoir 14 for storing a fluid including a target material, such as blood, a buffer reservoir 16 for storing a buffer solution, and a primary sensing material for binding or reacting with a target. First sensing material storage unit (18, 20), and the second sensing material storage unit for storing the secondary sensing material that can be secondarily coupled to the combined body formed by binding or reacting the primary sensing material ( 22).

In FIG. 1 or FIG. 2, two first sensing material storage units 18 and 20 are used. However, only one or three or more first sensing material storage units 18 and 20 may be used. As shown in FIG. 1, when two primary sensing materials, for example, primary sensing material A and primary sensing material B, are used, two targets can be analyzed in one analysis process. In other words, if the target storage contains antigen A and antigen B, the primary sensor A is antibody A which binds to antigen A, and the secondary sensor B is antibody B which binds to antigen B, Antigen A and antigen B can be detected.

The secondary sensing material is for detecting a target to which the primary sensing material is bound, and the secondary sensing material recombines to the target to which the primary sensing material is bound. When the target is to be detected by the intensity of fluorescence, the secondary sensing material may be a combination of fluorescent particles.

Each fluid reservoir 14, 16, 18, 20, 22 is connected to the main channel 24, and the end of the main channel is connected to the outlet 38.

The base 12 includes a plurality of flow controllers 26, 28, 30, 32 and microfluidic channel 40 to control the flow of fluid stored in the fluid reservoirs 14, 16, 18, 20, 22. A first actuator 34 and a second actuator 36 are provided for operation. The flow controller may include a first flow sensor 26 for controlling the flow of the target storage 14, a first flow sensor A 28 for controlling the flow of the first sensing material storage A 18, and a first flow controller 26 for controlling the flow of the first storage material A 18. The first sensing material flow controller B 30 for controlling the flow of the sensing material storage B 20, the second sensing material flow controller 32, etc. for controlling the flow of the second sensing material storage 22. It may include.

The flow controllers 26, 28, 30, 32, the first actuator 34 and the second actuator 36 may be operated in pneumatic manner and expanded to close the channel through which the fluid flows. Alternatively, the flow controllers 26, 28, 30, 32, the first actuator 34 and the second actuator 36 may be composed of piezoelectric elements.

2, the microfluidic analysis system according to the present invention includes a microfluidic analysis device 10, a flow controller 26, 28, 30, 32, and a first actuator 34 of the microfluidic analysis device 10. ) And a target detection signal acquisition unit 60 for acquiring a signal from the fluid flow controller 50 controlling the operation of the second actuator 36 and the target detection unit 42 performing target detection in the microfluidic channel 40. And an analysis device 70 for analyzing the detected signal.

When the flow controllers 26, 28, 30, 32 and the first actuator 34 and the second actuator 36 are pneumatically controlled, the fluid flow controller 50 may be a device that generates pneumatic pressure. When the target detector 42 generates the laser light and acquires the optical signal of the fluorescent particles excited by the laser light, the target detection signal acquisition unit 60 detects the optical signal that acquires the laser signal and the excited optical signal. It can be configured as a device.

Next, the microfluidic channel 40 for target capture according to a preferred embodiment of the present invention will be described.

3 is an exploded perspective view of a microfluidic channel for target capture according to a preferred embodiment of the present invention.

The microfluidic channel 30 includes a target capture part 100 provided with a plurality of fluid discharge holes 106 with the main channel 24 extended. In FIG. 3, the target capture unit 100 is provided with two of the first target capture unit 100a and the second target capture unit 100b, but in the practice of the present invention, the target capture unit 100 is captured. Of course, one or three or more may be provided depending on the type of target to be performed. On the other hand, the fluid discharge hole 106 is the place where the primary sensing material is seated, preferably the upper diameter of the fluid discharge hole 106 is larger than the diameter of the primary sensing material, so that the diameter decreases toward the lower diameter Good to do. In this way, the primary sensing material is not discharged to the lower portion of the fluid discharge hole 106 in a state where it is seated in the fluid discharge hole 106.

The membrane 102 is provided under the target capture unit 100. The membrane 102 is provided in a thin film form to open and close the lower portion of the target capture unit 100. To this end, a membrane operating part 104 for opening and closing the membrane 102 is provided under the membrane 102. Membrane actuator 104 may be comprised of a first membrane actuator 104a and a second membrane actuator 104b, the first membrane actuator 104a and the second membrane actuator 104b of FIG. It is operated by the first actuator 34 and the second actuator 36 in. As described above, the operation of the membrane actuator 104 may be controlled by pneumatic or piezoelectric elements.

Drainage channels 103a and 103b are formed between the lower portion of the target capture portion 100 and the membrane 102, and merge into the main channel 24 of the drainage channels 103a and 103b. That is, the fluid passing through the fluid discharge hole 106 of the target capture unit 100 is delivered to the main channel 24 through the drain channel formed on the membrane 102 and then discharged to the discharge port 38.

Meanwhile, the target capture unit 100, the membrane 102, and the membrane actuator 104 may be configured by a micro-electromechanical systems (MEMS) method. Referring to FIG. 3, the microfluidic channel 30 according to the present invention includes a first panel 110 having a target detector 42 attached thereto and a transparent panel, and a second panel forming a main channel 24. 122, a third panel 114 forming the membrane 102, and a fourth panel 116 constituting the membrane actuator 104.

In the above description regarding the control of the discharge of the fluid through the fluid discharge hole 106 formed in the target capture unit 100 has proposed a configuration employing the membrane 102, in addition to the membrane 102 in the practice of the present invention It is also possible to employ other configurations. For example, a fluid flow controller may be provided at a fluid discharge passage formed at a lower portion of the fluid discharge hole 106 so as to control fluid flow to control opening and closing of the fluid discharge passage. It is also possible to include a valve for manually or electronically controlling the opening and closing of the fluid discharge passage formed under the fluid discharge hole 106. This is because a feature of the present invention is to capture the target by controlling the fluid supplied through the main channel 24 to be discharged through the fluid discharge hole 106, thereby controlling the discharge of fluid through the fluid discharge hole 106 This is because various configurations can be used in place of the membrane 102. In the present invention, the fluid flow controller or the valve for controlling the discharge of the fluid through the fluid discharge hole 106 may be understood as a fluid discharge controller.

4 is a perspective view illustrating a configuration of a target detection unit in a microfluidic channel for target capture according to a preferred embodiment of the present invention.

The target detection unit 42 receives excitation light from the target detection signal acquisition unit 60 and irradiates the target to which the fluorescent material is attached, and emits light generated by excitation of the fluorescent material of the target. Transfer to the target detection signal acquisition unit 60. The target detection unit 42 may be provided with a double cladding optical fiber, as shown in FIG. 4, wherein the double cladding optical fiber has a core part 120 and a first cladding part formed while enclosing the core part 120 in a longitudinal direction. And a second cladding part 124 formed while enclosing the first cladding part 122 in the longitudinal direction. Unlike the ordinary optical fiber, the double cladding optical fiber can transmit and receive excitation light and emission light to one strand of optical fiber, and can detect fluorescence without using a fluorescence microscope. The structure of the fluid analysis system can be miniaturized and simplified. That is, the excitation light is transmitted through the core unit 120, and the light emitted from the excited fluorescent particles is transferred to the first cladding unit 122, thereby enabling the transmission of the excitation light and the light emission through one optical fiber.

In the double cladding optical fiber, the other end is preferably formed in the shape of the convex lens 126 or combined with the convex lens. Since the excitation light is collected at one point and reaches the secondary sensing material, the secondary sensing material is more effectively excited. Can be.

Next, the operation of the microfluidic channel for target capture according to a preferred embodiment of the present invention will be described.

5 to 10 are diagrams illustrating a process of capturing and detecting a target by using a microfluidic channel for target capture according to a preferred embodiment of the present invention.

Referring to FIG. 5, the first membrane 102a under the first target capture portion 100a is not in close contact with the second membrane 102b under the second target capture portion 100b. By 36, the state is in close contact with the lower portion of the second target capture portion 100b. In this state, the fluid including the primary sensing material A 110a stored in the first sensing material storage unit A 18 flows to the main channel 24. The fluid supplied to the main channel 24 passes through the fluid discharge hole 106 of the first target capture portion 100a and then drains to the first drain channel 103a. Accordingly, the first sensing material 110a is seated on the fluid discharge hole 106 of the first target capture portion 100a.

Next, referring to FIG. 6, the primary sensing material stored in the first sensing material storage B 20 after the pressure applied to the second membrane 102b is removed to return the second membrane 102b to its original position. Fluid including B (110b) flows to the main channel (24). Then, the primary sensing material B 110b is seated on the upper portion of the fluid discharge hole of the second target capture part 100b and the remaining fluid is drained to the second drain channel 103b.

Then, the buffer solution stored in the buffer storage unit 16 is transferred to the main channel 24 to wash the primary sensing substances not captured in the target capture unit 100.

Next, referring to FIG. 7, the targets 112a and 112b stored in the target storage unit 14 are flowed to the main channel 24 and a predetermined time is required for reaction or coupling with the primary sensing materials 110a and 110b. Incubate. The first target 112a is coupled to the primary sensing material A 110a and the second target 112b is coupled to the primary sensing material B 110b. Then, the buffer solution stored in the buffer storage unit 16 is again flowed to the main channel 24 to wash the unbound targets 112a and 112b.

Next, referring to FIG. 8, the fluid including the secondary sensing materials 112a and 112b is transferred from the secondary sensing material storage unit 22 to the main channel 24 and incubated for a predetermined time. The secondary sensing materials 114a and 114b are again coupled to the combination of the primary sensing materials 110a and 110b and the targets 112a and 112b. The buffer solution stored in the buffer storage unit 16 is flowed to the main channel 24 to wash the unbound secondary sensing materials 114a and 114b.

Next, referring to FIG. 9, the target detection unit 42 may detect the target by irradiating the excitation light to excite the secondary sensing material and obtaining the excited optical signal.

When the target detection is completed, as shown in FIG. 10, the first membrane 102a and the second membrane 102b are controlled to be in close contact with the lower portion of the first target capture portion 100a and the second target capture portion 100b and then buffered. The solution is flowed into the main channel 24 to remove particles from the target capture portion 100.

This process can be performed repeatedly, and it is also possible to continuously detect a plurality of targets by changing the type of target and the type of sensing material. Meanwhile, in the above description, the target capture unit 100 is continuously arranged along the main channel 24, but the main channel 24 is branched into a plurality of branches, and each branch has a target capture unit ( It is also possible to arrange one by one and control the flow of fluid to each branch to detect the target.

The present invention has been described above with reference to preferred embodiments. Those skilled in the art will understand that the present invention can be embodied in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown not in the above description but in the claims, and all differences within the scope should be construed as being included in the present invention.

1 is a perspective view of a microfluidic device for target capture according to a preferred embodiment of the present invention;

 2 is a block diagram of a microfluidic analysis system including a microfluidic device for target capture according to a preferred embodiment of the present invention;

3 is an exploded perspective view of a microfluidic channel for target capture according to a preferred embodiment of the present invention;

4 is a perspective view illustrating a configuration of a target detection unit in a microfluidic channel for target capture according to a preferred embodiment of the present invention;

5 to 10 are diagrams illustrating a process of capturing and detecting a target by using a microfluidic channel for target capture according to a preferred embodiment of the present invention.

<Description of the symbols for the main parts of the drawings>

10: microfluidic device 12: base

14: target storage unit 16: buffer solution storage unit

18, 20: first sensing material storage unit 22: second sensing material storage unit

24: main channel 26: target flow controller

28: first sensing material flow controller A 30: first sensing material flow controller B

32: second sensing material flow controller 34: first actuator

36: second actuator 38: outlet

40: microfluidic channel 42: target detector

50: fluid flow control unit 60: target detection signal acquisition unit

70: analysis apparatus 100, 100a, 100b: target capture unit

102, 102a, 102b: membrane zones 103a, 103b: drainage channels

104, 104a, 104b: membrane operating portion 106: fluid discharge hole

Claims (11)

Main channel; A target capture part formed in a part of the main channel and provided with a fluid discharge hole; A membrane which opens and closes the fluid discharge hole by being attached to and detached from a lower surface of the target capture unit; And A drain channel provided between the fluid discharge hole and the membrane to discharge the fluid passing through the fluid discharge hole Microfluidic channel for target capture comprising a. The method of claim 1, The fluid discharge hole is a microfluidic channel for target capture, characterized in that the size of the upper portion is larger than the size of the lower portion. The method of claim 1, And the membrane is operatively controlled by either pneumatic or piezoelectric elements. The method of claim 1, And a target detector configured to detect a target captured by the target capture unit at an upper portion of the target capture unit. The method of claim 4, wherein The target detection unit is composed of a double cladding optical fiber, the excitation light is transmitted to the core portion of the double cladding optical fiber, and the generated optical signal is transmitted to the first cladding portion surrounding the outside of the core portion for the target capture Microfluidic channels. The method of claim 5, A microfluidic channel for capturing a target, characterized in that a convex lens is molded or coupled to the end of the target detection unit. In a microfluidic channel device for target capture, A target storage unit for storing a target; A first sensing material storage unit storing a first sensing material primarily coupled to the target; A second sensing material storage unit storing a second sensing material which is secondarily coupled to the target; A flow controller for controlling the flow of the fluid stored in the target storage unit, the first sensing material storage unit, and the second sensing material storage unit; And Microfluidic channel according to any one of claims 1 to 6 Microfluidic channel device for target capture comprising a. In a microfluidic analysis system for target analysis, A target storage unit for storing a target; A first sensing material storage unit storing a first sensing material primarily coupled to the target; A second sensing material storage unit storing a second sensing material which is secondarily coupled to the target; A flow controller for controlling the flow of the fluid stored in the target storage unit, the first sensing material storage unit, and the second sensing material storage unit; Microfluidic channel according to any one of claims 1 to 6; A fluid flow controller for controlling the operation of the flow controller and the membrane; And A target detection signal acquisition unit for acquiring an optical signal according to the target detection Microfluidic analysis system for target analysis comprising a. The method of claim 8, Microfluid analysis system for target analysis, characterized in that further comprises an analysis device for analyzing the signal obtained by the target detection signal acquisition unit. Main channel; A target capture part formed in a part of the main channel and provided with a fluid discharge hole; And Including a fluid discharge control unit for controlling the discharge of the fluid through the fluid discharge hole of the target capture portion, And a target is captured in the fluid discharge hole when the fluid is controlled to be discharged through the fluid discharge hole. 11. The method of claim 10, The fluid discharge hole is a microfluidic channel for target capture, characterized in that the size of the upper portion is larger than the size of the lower portion.

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