KR102481011B1 - Lab-on-a-chip for field analysis using whole blood - Google Patents

Abstract

Disclosed is a lab-on-a-chip for in situ analysis which allows on-site diagnosis of a disorder or a disease or disease by separating plasma contained in whole blood and then contacting the plasma with a sample. To this end, the present invention comprises: an upper substrate including a filter unit which separates plasma from whole blood supplied to a blood inflow port, and a canal unit provided so that a plasma passage, through which the plasma separated through the filter unit moves, is exposed on a lower surface; a plasma inflow plate which is in close contact with a lower surface of the upper substrate to cover the plasma passage and has a plurality of inflow grooves formed on a contact surface of the plasma passage to allow the plasma flowing along the plasma passage to flow in; a lower substrate which is in close contact with a lower surface of the plasma inflow plate and has an open upper surface to form a vacuum room with an air intake space; and a blood inflow port sealing sheet which is installed in a passage communicating with the vacuum room and seals the vacuum room in a vacuum state so that suction force is generated in a direction of the vacuum room when the blood inflow port sealing sheet is detached. According to the present invention, it is possible to separate only plasma from whole blood using air pressure generated by a self-prepared vacuum room and produce a disposable biochip capable of detecting a specific disease by reacting separated plasma with a target sample.

Description

Lab-on-a-chip for field analysis using whole blood {LAB-ON-A-CHIP FOR FIELD ANALYSIS USING WHOLE BLOOD}

The present invention relates to a lab-on-a-chip for on-site analysis that can independently analyze plasma separated from whole blood as a sample without the aid of an external device, and more particularly, to separate plasma contained in whole blood and then contact the plasma with the sample. It relates to a lab-on-a-chip for on-site analysis that can diagnose diseases or diseases even in the field.

Lab-on-a-chip implements various laboratory procedures, such as sample separation, purification, mixing, labeling, analysis, and washing, on a small-sized chip. means to do In the design of this lab-on-a-chip, related technologies such as microfluidics and micro-LHS are mainly used.

In addition, in manufacturing a chip structure implementing microfluidic mechanics and a microfluidic manipulation system, a chip in which fine channels are formed inside the chip using semiconductor circuit design technology is on the market. For example, a microarray chip attaches a probe that reacts with a substance to be detected on a substrate at regular intervals, for example, a protein or DNA, and detects a substance that binds to the disease or disease. used for diagnosis of disease. A chip on which proteins are arranged is a microarray protein chip, and a chip on which DNA is arranged is a micro DNA chip.

Since analyte substances present in blood, such as proteins or antibodies, exist in very small amounts, attempts to increase detection efficiency by increasing the degree of integration of probes arrayed on a substrate have been continuously made. . As a result, a nanoarray chip with increased integration to the nano level has recently been reported.

In general, analysis of fluid samples is mainly used in the field of chemistry and biotechnology, as well as analysis of blood and body fluids collected from patients in clinical practice and diagnosis of diseases through this analysis. In this way, detecting and analyzing a small amount of an analyte contained in a fluid sample such as blood, body fluid, or urine is an antigen or It includes analyzing the reaction with proteins such as antibodies, DNA/RNA, receptors, or other substances through detection signals using optical means such as fluorescent materials or electrical means.

In the analysis of a fluid sample, analyzing a very small amount of a sample is very important for a biochip including a lab-on-a-chip. In general, a lab-on-a-chip for analyzing a fluid sample includes a sample injection unit for providing a fluid sample to the chip, a filtering unit using a paper filter, etc. to remove noise substances other than analyte substances in the fluid, and A reaction in which a substance binds to a detection signal generating substance, such as a fluorescent material, and a reaction between an analyte bound to the detection signal substance and a substance immobilized on a substrate that specifically binds to the analyte. and a detection unit that analyzes a detection signal of an analyte such as a protein present in blood, urine, or body fluid using a chip.

On the other hand, whole blood contains plasma (46-63%), red blood cells (diameter: 7-8 μm, thickness: 2 μm, 50% of blood), white blood cells (diameter: 10-20 μm) and platelets (diameter: 2 μm). It is composed of hemocytes (37-54%) containing 1-3 μm). In blood, in addition to plasma and blood cell components, proteins, antigens or antibodies, ligands, receptors, etc. exist as various types of analyte substances that can be used for diagnosis of diseases or diseases.

Accordingly, various methods and devices for detecting a disease or an analyte related to a disease from whole blood have been developed.

Recently, research has been conducted to quickly obtain various information from terminal blood collected clinically by using lab-on-a-chip for blood analysis, and as a result, rapid-chip or rapid-kit has been developed. It became.

However, the blood analysis (rapid) chip according to the prior art essentially includes a filtering step. Such filtering, in particular, filtering using a paper filter causes a dead volume of the fluid sample due to absorption of the fluid sample by the filter. As a result, a minimum sample injection amount (100 μl or more in the case of blood) is required to detect and analyze the analyte present in the sample, which makes it impossible to detect and analyze the analyte present in a very small amount of fluid sample. . Accordingly, research on microfluidics to replace paper filters continues, but until now, the dead volume of fluid samples has not been significantly reduced due to limitations in separation efficiency.

Republic of Korea Patent Registration No. 10-2114446 (2020.05.21 announcement) Republic of Korea Patent Registration No. 10-0912530 (2009.08.18 announcement) Republic of Korea Patent No. 10-2244375 (2021.04.27 announcement) Republic of Korea Patent Registration No. 10-0843339 (Announced on July 3, 2008) Republic of Korea Patent Registration No. 10-0860075 (Announced on September 24, 2008)

Therefore, an object of the present invention is for on-site analysis that can separate plasma from whole blood on its own without the help of an external device, and can naturally react the plasma separated from whole blood with a sample to secure the basis for diagnosing a target disease or disease. It is providing lab-on-a-chip.

In order to achieve the above object of the present invention, in one embodiment of the present invention, a filter unit for separating plasma from whole blood supplied to the blood inlet port, and a plasma passage through which the separated plasma moves through the filter unit are exposed on the lower surface. An upper substrate including a canal portion provided therein, and a plasma inlet plate adhered to the lower surface of the upper substrate to cover the plasma passage and having a plurality of inlet grooves formed on the contact surface of the plasma passage so that plasma flowing along the plasma passage is introduced; , a lower substrate in close contact with the lower surface of the plasma inlet plate and having an open upper surface to form a vacuum room having an air suction space, and a vacuum installed in a passage communicating with the vacuum room so that a suction force is generated in the direction of the vacuum room during detachment. Provided is a lab-on-a-chip for on-site analysis including a blood inlet sealing sheet for sealing a vacuum room in the present state.

According to the present invention, only plasma can be separated from whole blood using air pressure generated by a self-prepared vacuum room, and a disposable biochip that detects a specific disease can be manufactured by reacting the separated plasma with a target sample.

In addition, since the present invention can be manufactured through an injection process, the price is low and mass production is possible, and laboratory tests for diseases can be performed in a small clinic, field or home environment.

In addition, the present invention moves whole blood from which plasma is separated into the main passage, moves the plasma separated from the whole blood to the plasma canal adjacent to the main passage, and through a plurality of plasma inlet channels formed between the main passage and the plasma canal. Plasma can be moved from whole blood moving along the path of the main passage to the plasma canal, so that plasma can be separated from whole blood without obstruction due to accumulation of blood cells in a specific section.

Moreover, since plasma separated from whole blood contains a minimum amount of red blood cells, the present invention can prevent problems such as slowing down the reaction due to hemoglobin, and can use fewer expensive reagents.

1 is a perspective view illustrating a lab-on-a-chip for on-site analysis according to the present invention.
2 is an exploded perspective view illustrating a lab-on-a-chip for on-site analysis according to the present invention.
3 is a plan view showing an upper substrate according to the present invention.
4 and 5 are partially enlarged plan views for explaining the plasma inflow channel provided between the blood canal and the plasma canal in the upper substrate according to the present invention.
6 is a partially enlarged plan view for explaining the inlet groove of the plasma inlet plate in contact with the plasma passage of the present invention.
7 is a cross-sectional view showing a lab-on-a-chip for in situ analysis according to the present invention.

Hereinafter, a lab-on-a-chip (hereinafter, abbreviated as 'lab-on-a-chip for in situ analysis') using whole blood according to preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

1 is a perspective view illustrating a lab-on-a-chip for in situ analysis according to an embodiment of the present invention, and FIG. 2 is an exploded perspective view illustrating a lab-on-a-chip for in situ analysis according to an embodiment of the present invention.

1 and 2, the lab-on-a-chip for on-site analysis according to the present invention includes a filter unit 110 that separates plasma from whole blood supplied to the blood inlet 113 and a plasma passage 122 that moves the plasma. An upper substrate 100 including a canal portion 120 having a canal portion 120 provided on the lower surface of the upper substrate 100 and a plurality of blood plasma flowing in the horizontal direction along the plasma passage 122 are partially introduced downward. Plasma inlet plate 200 with two inlet grooves 210 and a lower substrate provided with a vacuum room 310 provided on the lower surface of the plasma inlet plate 200 to suck in air from the outside when the sealed state is released 300, and a blood inlet sealing sheet 400 installed in a passage communicating with the vacuum room 310 to seal the vacuum room 310 in a vacuum state. At this time, the target reagent may be provided in the inlet groove 210 or injected into the blood inlet 113 together with whole blood.

On the other hand, the upper substrate 100 shown in FIG. 2 has a composition in which the lower surface is exposed, unlike the plasma inlet plate 200 and the lower substrate 300, in which the upper surface is exposed, so that the characteristics of the lower surface can be clearly revealed.

Hereinafter, each component will be described in more detail with reference to the drawings.

3 is a plan view showing an upper substrate according to the present invention.

Referring to FIGS. 1 to 3 , the lab-on-a-chip for in situ analysis according to the present invention includes an upper substrate 100 .

The upper substrate 100 separates plasma included in the whole blood from the whole blood while moving the whole blood supplied from the outside, and a filter unit 110 for separating plasma from the whole blood supplied to the blood inlet 113, and the filter The canal part 120 is included so that the plasma passage 122 through which the plasma separated through the part 110 moves is exposed on the lower surface.

The filter unit 110 includes a first body 111 having a blood canal 112 through which blood can flow, a blood inlet 113 for supplying blood introduced from the outside to the blood canal 112, The plasma canal 114 and 115 formed in the first body 111 along the blood canal 112, and the blood canal 112 and the plasma canal 114 and 115 in the first body 111 so that blood cells do not pass but blood plasma passes It includes a plurality of plasma inlet channels 116 installed between them. If necessary, the filter unit 110 according to the present invention may further include a plasma receiving port communicating with the rear end of the plasma canal 114 or 115 and accommodating plasma passing through the plasma canal 114 or 115 .

The first body 111 constituting the filter unit 110 according to the present invention provides a space in which the blood canal 112, the plasma canal 114 and 115, the plasma inflow channel 116, and the plasma receptor are formed. In the first body 111, spaces other than the blood canal 112, the plasma canal 114 and 115, the plasma inflow channel 116, and the plasma receptor may be formed as a closed space.

The blood canal 112 may be formed in a zigzag structure in the shape of 'D' or 'S' so as to have the longest length on the first body 111.

As shown in FIG. 3, the blood canal 112 may be composed of a front end to which the plasma inflow channel 116 is not connected and a rear end to which the plasma inflow channel 116 is connected, and suction power is provided from the plasma inflow channel 116. It may be formed to have a long length so that blood does not pass through the front end before becoming.

Specifically, the front end of the blood canal 112 includes 100 to 140 lines horizontally along the longitudinal direction, and the rear end of the blood canal 112 includes 100 to 140 lines horizontally along the longitudinal direction. .

As a specific aspect, the blood canal 112 according to the present invention may be formed by being dug in a hemispherical shape at the lower end of the first body 111 to be exposed to the outside of the first body 111. It may be formed in a tubular shape inside the first body 111 so as not to be exposed to the outside. At this time, the blood canal 112 is isolated from the outside by the plasma inlet plate 200 even though it is formed by being dug into a hemispherical shape at the lower end of the first body 111, so it communicates with the blood inlet 113 from the blood inlet 113. The introduced whole blood can be moved along the longitudinal direction.

Specifically, the first body 111 may have a width of 20 to 40 mm, a thickness of 1,000 μm to 1,400 μm, and a length of 30 to 80 mm.

In addition, the blood canal 112 may have a width of 30 to 60 μm and a depth of 10 to 20 μm so that blood can flow smoothly. This is because the size of red blood cells is generally 7 to 8 μm. Here, the depth means the length of the first body 111 in the thickness direction.

Meanwhile, the first body 111 may be formed of any one of plastic polymer, silicon, glass, or rubber, and is preferably formed of a cheap plastic polymer so that it can be used in a disposable biochip.

The plastic polymer includes cycloolefin copolymer, poly-dimethyl siloxane, polymethylmethacrylate, polycarbonate, polyamide, polyethylene , polypropylene, polyphenylene ether, polystyrene, polyoxymethylene, polyetheretherketone, polytetrafluoroethylene, polyvinylchloride ), polyvinylidene fluoride, polybutyleneterephthalate, fluorinated ethylenepropylene, and perfluoralkoxyalkane. Any one selected from the group consisting of may be used.

The blood inlet 113 constituting the filter unit 110 according to the present invention is provided at the front end of the blood canal 112 and communicates with the blood canal 112 . The blood inlet 113 provides a function of receiving whole blood introduced from the outside and then supplying it to the blood canal 112, and may have an inner diameter in the range of 250 μm to 3.000 μm.

The blood inlet 113 may be formed in the form of a recessed groove in the first body 111 or may be formed in the form of a through hole in the first body 111 . For example, when the blood canal 112 is formed inside the first body 111, the blood inlet 113 is formed in the form of a recessed groove so that the blood canal 112 is exposed to the outside of the first body 111. When formed, the blood inlet 113 is formed in the form of a through hole.

As a specific aspect, the blood inlet 113 according to the present invention may be formed to have a structure in which the cross-sectional area is gradually reduced as it progresses from the top to the bottom. This is to reduce the difficulty of supplying blood and to prevent the blood supplied from the outside from being separated from the blood inlet 113 .

The plasma canal 114 and 115 constituting the filter unit 110 according to the present invention are disposed along the blood canal 112 to provide a movement path for the plasma separated from the whole blood, and the first one is spaced apart from the blood canal 112. It is formed on the body 111.

The plasma canal 114 and 115 may be formed at the lower end of the first body 111 to be exposed to the outside of the first body 111 along the blood canal 112, or may not be exposed to the outside of the first body 111. It may be formed inside the first body 111 so that it does not.

Specifically, the plasma canal 114 and 115 may have a width of 20 to 40 μm and a depth of 10 to 20 μm for smooth movement of blood plasma and convenience of manufacturing.

In a specific embodiment, the plasma canal 114 and 115 according to the present invention include the first plasma canal 114 provided on one side of the blood canal 112 and the blood canal 112 as the center. It may be composed of a second plasma canal 115 provided on the other side of the blood canal 112 opposite to one side. This is to provide the plasma inflow channel 116 communicating with the plasma canal 114 and 115 on both left and right sides of the blood canal 112.

For example, as shown in FIG. 3, the first plasma canal 114 may be formed in a zigzag structure in the shape of 'D' or 'S', and the second plasma canal 115 is shown in FIG. As described above, it may be formed in a zigzag structure in the form of 'L' or 'S'.

The first plasma canal 114 includes 100 to 140 lines horizontally along the longitudinal direction, and the second plasma canal 115 includes 100 to 140 lines horizontally along the longitudinal direction. At this time, it is preferable that the first plasma canal 114 and the second plasma canal 115 are provided in the same number in the first body 111 .

4 and 5 are partially enlarged plan views for explaining the plasma inflow channel provided between the blood canal and the plasma canal in the upper substrate according to the present invention.

4 and 5, the plasma inlet channel 116 constituting the filter unit 110 according to the present invention is the blood canal 112 and the plasma canal 114 and 115 along the longitudinal direction of the blood canal 112. Plasma separated from the whole blood transported along the blood canal 112 is introduced into the plasma canal 114 and 115 by installing a plurality of them in between.

The plasma inlet channel 116 is formed to have a cross-sectional area through which blood cells included in the whole blood, particularly red blood cells, cannot pass through, thereby allowing plasma included in the whole blood to pass through and limiting the movement of the blood cells included in the whole blood.

The plasma inflow channel 116 may be formed at the lower end of the first body 111 so as to be exposed to the outside of the first body 111, or the first body 111 not exposed to the outside of the first body 111. ) may be formed inside.

Specifically, the plasma inflow channel 116 may have a width of 4 to 6 μm and a depth of 2 to 3 μm, preferably 2.5 μm so as to be manufactured by an injection process. This is because the size of red blood cells is generally 7 to 8 μm and the minimum thickness exceeds 2.5 μm.

The plasma inflow channel 116 may be formed in a polygonal column shape such as a cylindrical shape, a triangular columnar shape, a square columnar shape, or a pentagonal columnar shape, but is not limited thereto.

The plasma inflow channel 116 may be provided at right angles between the blood canal 112 and the plasma canal 114 and 115 to maximize the number of installations for convenience of manufacturing.

Since the plasma inlet channel 116 is continuously installed along the longitudinal direction of the blood canal 112, which is a single blood transfer path, the plasma inlet channel 116 installed at the front end of the blood canal 112 is closed by blood cells. However, since other plasma inflow channels 116 installed along the transfer direction of the blood exist, the plasma contained in the blood can be transferred to the plasma channels 114 and 115.

1,500 to 3,000 plasma inlet channels 116 may be provided on both sides of any one line among a plurality of horizontally arranged lines constituting the blood canal 112 along the longitudinal direction of the first body 111. there is.

The plasma receptor constituting the filter unit 110 according to the present invention communicates with the rear end of the plasma canal 114 and 115 and is a space in which plasma passing through the plasma canal 114 and 115 is accommodated, and biological information about a biological sample is stored as a disposable biochip. Store the plasma used for detection.

The plasma receptor may be formed at the lower end of the first body 111 so as to be exposed to the outside of the first body 111, or inside the first body 111 so as not to be exposed to the outside of the first body 111. may be formed.

The filter unit 110 according to the present invention may further include a blood receiving port (not shown).

The blood receiving port communicates with the rear end of the blood canal 112 and is a space in which blood passing through the blood canal 112 is accommodated, and plasma is separated and blood containing blood cells is stored.

The blood container may be formed at the lower end of the first body 111 so as to be exposed to the outside of the first body 111, or inside the first body 111 so as not to be exposed to the outside of the first body 111. may be formed.

Meanwhile, the canal part 120 constituting the upper substrate 100 according to the present invention communicates with the plasma canal 114 and 115 of the filter part 110 or the plasma receiving port, and the plasma separated through the filter part 110 The moving plasma passage 122 is exposed on the lower surface of the second body 121 so as to come into contact with the plasma inlet plate 200 .

More specifically, the canal portion 120 includes a second body 121 extending from the first body 111 and a blood plasma passage 122 provided in the second body 121.

Since the second body 121 is integrally formed with the first body 111, it is preferable to have the same width and thickness as the first body 111. Specifically, the second body 121 may have a width of 20 to 40 mm, a thickness of 500 μm to 1,500 μm, and a length of 30 to 80 mm.

The blood plasma passage 122 may be formed in a zigzag structure in the shape of 'D' or 'S' so as to have a maximum length on the second body 121 . The blood plasma passage 122 may have a width of 150 to 200 μm and a depth of 20 to 30 μm so that blood plasma can move smoothly.

If necessary, the canal part 120 may be provided with a residual plasma inlet 124. The residual plasma inlet 124 is provided at the rear of the plasma passage 122 so as to communicate with the end of the plasma passage 122, and cannot move to the inlet groove 210 of the plasma inlet plate 200 and the plasma passage 122 ) provides a passage through which residual plasma transferred to the end of the plasma inlet plate 200 is discharged. Specifically, the residual plasma inlet 124 may be formed in the shape of a recessed groove at the rear end of the second body 121 to be exposed on the upper surface of the plasma inlet plate 200 .

Also, the upper substrate 100 may be provided with a plurality of upper assembly holes 130 through which fastening means 600 such as bolts are inserted so that they can be combined with the plasma inlet plate 200 without using an adhesive. Specifically, the upper assembly hole 130 may be formed along the edges of the first body 111 and the second body 121 as shown in FIGS. 1 and 2 .

In addition, the upper substrate 100 may be provided with one or more upper fixing holes 140 through which fastening means 600 such as bolts are interpolated so that a porous polymer mass described later can be fixed to the upper substrate 100 . Specifically, the upper fixing hole 140 may be provided around the plasma passage 122 as shown in FIGS. 1 and 2 .

In addition, the upper substrate 100 shortens the movement path of the air stored in the vacuum room 310 to form the vacuum room 310 in a vacuum state so that the air in the vacuum room 310 can be easily discharged to the outside. A first intake hole 150 may be provided. The first intake hole 150 is provided on the upper substrate 100 to pass through the upper substrate 100 in a vertical direction.

6 is a partially enlarged plan view illustrating the inlet groove of the plasma inlet plate in contact with the plasma passage of the present invention, and FIG. 7 is a cross-sectional view showing a lab-on-a-chip for on-site analysis according to the present invention.

Referring to FIGS. 1 and 2 , the lab-on-a-chip for in situ analysis according to the present invention includes a plasma inlet plate 200.

The plasma inlet plate 200 is closely adhered to the lower surface of the upper substrate 100 to cover the plasma passage 122, and blood plasma flowing along the plasma passage 122 of the canal part 120 as shown in FIGS. 6 and 7 A plurality of inlet grooves 210 are formed on the contact surface of the plasma passage 122 to be sucked downward through the plasma passage 122 by this vacuum or atmospheric pressure difference.

The plasma inlet plate 200 adheres to the lower surface of the upper substrate 100 to seal the blood canal 112, the plasma canal 114 and 115, the plasma inlet channel 116, the plasma receptor, and the plasma passage 122 from the outside. and is formed to have an area capable of covering the lower portion of the upper substrate 100.

Specifically, the plasma inlet plate 200 is formed to have the same width and the same length as the upper substrate 100, and may have a thickness in the range of 2,000 to 3,000 μm.

The plasma inlet plate 200 may be attached to the upper substrate 100 using any one of a bolt assembly method, a hot bonding method, an epoxy bonding method, a chemical bonding method, an ultrasonic bonding method, and a plasma bonding method.

Also, as shown in FIG. 6 , the plurality of inlet grooves 210 may be configured to form two or more lines, preferably two to five lines, on a surface in contact with the blood plasma passage 122 . In other words, the plasma passage 122 may be formed to be in contact with one line of inlet grooves 210 along the length direction, but contact with two or more rows of inlet grooves 210 to lower the manufacturing cost by lowering the difficulty of processing. It is desirable that it is formed so as to be.

As one embodiment, the inlet groove 210 according to the present invention is two or three inlet grooves 210 to form two or three lines on the contact surface with the plasma passage 122 as shown on the left side of FIG. constitutes the inlet groove cluster, and the inlet groove cluster and the inlet groove cluster are formed to be spaced apart at predetermined intervals.

As another embodiment, in the inlet groove 210 according to the present invention, one line is formed at regular intervals as shown on the right side of FIG.

In addition, the inflow groove 210 may be formed to have an inner diameter gradually reduced as it progresses from the upper surface of the plasma inlet plate 200 to the lower surface. This is to improve the convenience of processing compared to a structure having the same inner diameter.

Specifically, the inlet groove 210 may have an inner diameter of 45 μm to 55 μm and a depth of 55 μm to 65 μm.

The plasma inlet plate 200 may be made of at least one of polydimethylsiloxan (PDMS), polyurethane, polystyrene, and polyorganosiloxane providing gas permeability. In other words, when the blood inlet sealing sheet 400 is detached from the upper substrate 100, the plasma inlet plate 200 in contact with the upper substrate 100 provides a gradual suction force to the upper substrate 100 in the entire section, The plasma moving along the passage 122 is sucked into the plurality of inlet grooves 210 .

In this way, when the plasma inlet plate 200 is made of a material providing gas permeability, the blood inlet 113 and the first intake hole 150 are formed through the blood inlet sealing sheet 400 and the intake hole sealing sheet 500. Even if it is sealed, a problem in which external air is sucked through the side surface of the plasma inlet plate 200 may occur. Accordingly, in the wrap-on-a-chip according to the present invention, a sealing film (not shown) may be attached to the side of the plasma inlet plate 200 to block air flow so that air does not pass through. At this time, a polymer film, double-sided tape, etc. may be used as the sealing film.

If necessary, in the wrap-on-a-chip according to the present invention, a sealing film may be provided to cover not only the side surface of the plasma inlet plate 200 but also the side surfaces of the upper substrate 100 and the lower substrate 200.

In addition, the plasma inlet plate 200 may be provided with a residual plasma discharge hole 220 facing the residual plasma inlet 124 and communicating with the vacuum room 310 . The residual plasma discharge hole 220 provides a function of moving the residual plasma discharged from the residual plasma inlet 124 to the vacuum room 310 .

In addition, the plasma inlet plate 200 is provided with a plurality of intermediate assembly holes 230 for inserting a fastening means 600 such as a bolt so that it can be combined with the upper substrate 100 and the lower substrate 300 without using an adhesive. It can be. Specifically, the middle assembly hole 230 is formed so that its upper surface faces the upper assembly hole 130 and its lower surface faces the side plate 330 of the lower substrate 300 .

In addition, the plasma inlet plate 200 may be provided with one or more intermediate fixing holes 240 for inserting a fastening means 600 such as a bolt so as to fix the porous polymer mass to the plasma inlet plate 200. Specifically, the middle fixing hole 240 may be provided around the inlet groove 210 as shown in FIGS. 1 and 2 .

In addition, the plasma inlet plate 200 shortens the moving path of the air stored in the vacuum room 310 to form the vacuum room 310 in a vacuum state, so that the air in the vacuum room 310 can be easily discharged to the outside. A second intake hole 250 may be provided. The second intake hole 250 is provided on the plasma inlet plate 200 so as to penetrate the plasma inlet plate 200 in the vertical direction, and is formed to face the first intake hole 150 described above, and the vacuum room 310 ) is formed to be in communication with.

As such, when the first intake hole 150 and the second intake hole 250 are not provided, in order to manufacture the lab-on-a-chip for on-site analysis of the present invention, a suction pump is connected to the blood inlet 113 and then the blood canal 112, the plasma inlet channel 116, the plasma canal 114, 115, the plasma passage 122, and the plasma inlet plate 200, because the air stored in the vacuum room 310 must be discharged through a complicated and long passage. It takes a relatively long time to convert the room 310 to a vacuum state, which may cause a decrease in productivity.

Even if the lab-on-a-chip for on-site analysis of the present invention is equipped with a residual plasma inlet 124 and a residual plasma outlet hole 220, the blood canal 112, plasma inlet channel 116, plasma canal 114,115, and plasma passage 122, the residual plasma inlet 124, and the residual plasma discharge hole 220, the air stored in the vacuum room 310 must be discharged, so it takes a long time to convert the vacuum room 310 to a vacuum state. .

On the other hand, when the first intake hole 150 and the second intake hole 250 are provided, the vacuum room 310 and the outside are connected through the first intake hole 150 and the second intake hole 250, In order to manufacture the lab-on-a-chip for on-site analysis of the present invention, after connecting a suction pump to the first intake hole 150, air stored in the vacuum room 310 is supplied through the first intake hole 150 and the second intake hole 250. ) It is possible to convert the vacuum room 310 to be discharged through a vacuum state within a few seconds. Accordingly, in the present invention, the case where the first intake hole 150 and the second intake hole 250 are provided is several times more than the case where the first intake hole 150 and the second intake hole 250 are not provided. efficiency can be provided.

Referring to FIGS. 1 and 2 , the lab-on-a-chip for in situ analysis according to the present invention includes a lower substrate 300 .

The lower substrate 300 is in close contact with the lower surface of the plasma inlet plate 200, and the upper surface is opened to form a vacuum room 310 having an air intake space.

The lower substrate 300 may be composed of a bottom plate 320 and a side plate 330 formed in a vertical direction along an edge of the bottom plate 320 to form a vacuum room 310 . This vacuum room 310 maintains a vacuum state and generates a suction force in the direction of the vacuum room 310 when communicating with the atmosphere, and sequentially the residual plasma discharge hole 220, the residual plasma inlet 124, and the plasma passage 122 ), the plasma canal 114 and 115, the plasma inflow channel 116, the blood canal 112, and the blood inlet 113.

The bottom plate 320 is formed to have the same width and length as the plasma inlet plate 200, and may have a thickness ranging from 0.5 mm to 3.0 mm.

The base plate 320 may be formed of inexpensive plastic polymer, thermally conductive plastic, or metal such as aluminum having a thermal conductivity of 15 to 400 W/mK so as to be used in a disposable biochip.

Here, the thermally conductive plastic refers to a plastic having excellent thermal conductivity, and has excellent thermal conductivity by increasing the heat transfer rate by 5 to 100 times compared to general plastics. In addition, thermally conductive plastics are properties of plastics that enable very precise injection molding, have excellent corrosion resistance and sufficient mechanical strength, and at the same time have excellent thermal conductivity. It is a material with improved properties. Specifically, the thermal conductivity of the thermally conductive plastic is about 1 to 20W/mK, and has a very high thermal conductivity compared to the thermal conductivity of 0.25W/mK of general plastics. As such, thermally conductive plastics have very high thermal conductivity compared to general plastics, but rather low thermal conductivity compared to metal materials.

For example, the thermally conductive plastic may consist of a mixture of 30% to 70% graphite by weight and 30% to 70% plastic by weight.

The side plate 330 is formed in the vertical direction at the front end, rear end, left end, and right end of the bottom plate 320, respectively, and has a height in the range of 1,500 μm to 2,500 μm, and a thickness in the range of 500 μm to 5,000 μm. can be formed as

Such a side plate 330 may be formed of inexpensive plastic polymer, thermally conductive plastic, or metal such as aluminum having a thermal conductivity of 15 to 400 W/mK so as to be used in a disposable biochip.

The plastic polymers include cycloolefin copolymer, polydimethylsiloxane, polymethylmethacrylate, polycarbonate, polyamide, polyethylene, polypropylene, polyphenylene ether, polystyrene, polyoxymethylene, polyetheretherketone, polytetra Any one selected from the group consisting of propylene, polyvinyl chloride, polyvinylidene fluoride, polybutylene terephthalate, fluorinated ethylene propylene, and perfluoroalkoxyalkane may be used.

In other words, in the lower substrate 300, both the bottom plate 320 and the side plate 330 may be made of metal having a thermal conductivity of 15 to 400 W/mK, and both the bottom plate 320 and the side plate 330 may be made of plastic polymer. However, the base plate 320 may be formed of a metal having a thermal conductivity of 15 to 400 W/mK, and the side plate 330 may be formed of a plastic polymer or a thermally conductive plastic.

Processing of the above-described upper substrate 100, plasma inlet plate 200, and lower substrate 300 may be performed in various ways depending on the material, for example, hot embossing, injection molding, or the like. ), NC (Numerical Control) machining, laser ablation, electric discharge, casting, stereolithography, rapid prototyping, and photolithography. It can be formed using one method. In particular, when forming a plastic polymer for application to an inexpensive disposable biochip, it is preferable to use a hot embossing or injection molding method.

The lower substrate 300 may be provided with a plurality of lower assembly holes 340 in the side plate 330 or a flange protruding along the edge of the side plate 330 . The plurality of lower assembly holes 340 are spaces where fastening means 600 such as bolts are assembled so that the lower substrate 300 can be coupled to the upper substrate 100 and the plasma inlet plate 200 without using an adhesive. provides At this time, the plurality of lower assembly holes 340 are formed to face the middle assembly grooves 220 of the plasma inlet plate 200.

Referring to FIGS. 1 and 2 , the lab-on-a-chip for on-site analysis according to the present invention may further include an intake hole sealing sheet 500 .

The intake hole sealing sheet 500 seals the first intake hole 150 of the upper substrate 100 so that the vacuum room 310 maintains a vacuum state, and as shown in FIG. 1, the first intake hole 150 is It is attached to the upper surface of the formed upper substrate 100 .

The intake hole sealing sheet 500 converts the vacuum room 310 into a vacuum state through the first intake hole 150 and the second intake hole 250, and then the first intake hole 150 and the second intake hole (250) provides a function to block from the outside so that it is not used.

As a specific aspect, the intake hole sealing sheet 500 according to the present invention may be formed of a polymer film or the like through which air does not pass.

Here, an adhesive or double-sided tape is exemplified as a sealing means capable of attaching the intake hole sealing sheet 500 to the upper substrate 100, but other sealing means capable of permanently sealing the first intake hole 150. may be replaced with

Referring to FIG. 1 , the lab-on-a-chip for in situ analysis according to the present invention may further include a porous polymer mass 700 .

The porous polymer mass 700 is filled in the vacuum room 310 of the lower substrate 300 to provide gas permeation characteristics. According to these characteristics, when the blood inlet sealing sheet 400 is removed from the lab-on-a-chip for on-site analysis by the user and the vacuum state of the vacuum room 310 is converted to the atmospheric pressure state, the vacuum room 310 in the vacuum state is at atmospheric pressure. By delaying the transition to the state too quickly, it is possible to secure time to smoothly perform diagnostic activities using lab-on-a-chip.

The porous polymer mass 700 is a vacuum room so that the lower substrate 300 can continue to function as a suction pump until the process of separating plasma from whole blood and the process of moving plasma to the inlet groove 210 are completed. Adjust the suction power of (310).

The porous polymer mass 700 may be composed of at least one of polydimethylsiloxane (PDMS), polyurethane, polystyrene, and polyorganosiloxane providing gas permeability.

The porous polymer mass 700 may be provided with one or more lower fixing holes 710 for inserting fastening means 600 such as bolts assembled through the middle fixing holes 240 of the plasma inlet plate 200. . At this time, the lower fixing hole 710 may be formed to face the middle fixing hole 240 so that the fastening means 600 can be smoothly assembled.

Referring to FIGS. 1 and 2 , the lab-on-a-chip for on-site analysis according to the present invention includes a sealing sheet 400 for a blood inlet.

The blood inlet sealing sheet 400 is installed in a passage communicating with the vacuum room 310, and seals the vacuum room 310 in a vacuum state so that a suction force is generated in the direction of the vacuum room 310 when detached by the user. .

More specifically, when the blood inlet sealing sheet 400 is attached to the upper surface of the upper substrate 100 to seal the blood inlet 113 of the upper substrate 100, the blood inlet sealing sheet 400 is the wrap-on-a-word of the present invention. All internal spaces of the chip are sealed.

When the blood inlet sealing sheet 400 is detached by the user, the movement of the whole blood supplied to the blood inlet 113 is promoted by the suction force generated from the lower substrate 300 .

As a specific aspect, the blood inlet sealing sheet 400 according to the present invention may be formed of a polymer film that does not allow air to pass through, and the blood inlet 113 is sealed through an adhesive or double-sided tape, as shown in FIG. It is attached to the upper surface of the upper substrate 100 on which the inlet 113 is formed.

Here, an adhesive or double-sided tate is exemplified as a sealing means capable of attaching the blood inlet sealing sheet 400 to the upper substrate 100, but after sealing the blood inlet 113, the blood inlet sealing sheet ( 400) may be replaced with other sealing means capable of detaching from the upper substrate 100.

Meanwhile, a plurality of plasma inlet channels 116 are formed along the longitudinal direction of the blood canal 112, and when the blood inlet sealing sheet 400 is removed, the plasma inlet channel 116 is connected to the rear end of the blood canal 112. The suction force transmitted to the blood canal 112 through ) provides stronger strength than the suction force transmitted to the blood canal 112 through the plasma inlet channel 116 connected to the front end of the blood canal 112.

As such, the suction force transmitted to the blood canal 112 moves the whole blood accommodated in the blood inlet 113 to the rear end of the blood canal 112, and the suction force transmitted to the plasma inlet channel 116 moves the blood canal 112 Plasma is separated from whole blood flowing along and moved to the plasma canal 114 and 115, and the suction power transmitted to the plasma canal 114 and 115 moves the plasma to the rear end of the plasma canal 114 and 115.

In other words, the lab-on-a-chip for on-site analysis according to the present invention can separate plasma from whole blood and react the plasma with a target reagent without using a device such as a suction pump.

Although the above has been described with reference to preferred embodiments of the present invention, those skilled in the art can variously modify and change the present invention without departing from the spirit and scope of the present invention described in the claims below. You will understand that you can.

100: upper substrate 110: filter unit
111: first body 112: blood canal
113: blood inlet 114: first plasma canal
115: second plasma canal 116: plasma inflow channel
120: canal part 121: second body
122: plasma passage 124: residual plasma inlet hole
130: upper assembly hole 140: upper fixing hole
150: first intake hole 200: plasma inlet plate
210: inflow groove 220: residual plasma discharge hole
230: Intermediate assembly hole 240: Intermediate fixing hole
250: second intake hole 300: lower substrate
310: vacuum room 320: bottom plate
330: side plate 400: blood inlet sealing sheet
500: intake hole sealing sheet 600: fastening means
700: porous polymer mass 710: lower fixing hole

Claims (12)

an upper substrate including a filter unit for separating plasma from whole blood supplied to the blood inlet port, and a canal unit provided to expose a plasma passage through which the plasma separated through the filter unit moves on the lower surface;
a plasma inlet plate adhered to the lower surface of the upper substrate to cover the plasma passage and having a plurality of inlet grooves formed on a contact surface of the plasma passage so that plasma flowing along the plasma passage is introduced;
a lower substrate adhered to the lower surface of the plasma inlet plate and having an open upper surface to form a vacuum room having an air intake space;
a blood inlet sealing sheet installed in a passage communicating with the vacuum room to seal the vacuum room in a vacuum state so that a suction force is generated in the direction of the vacuum room when detached; and
A lab-on-a-chip for on-site analysis comprising a porous polymer mass filled in a vacuum room to provide gas permeability so as to delay the process of converting the vacuum room to atmospheric pressure. The method of claim 1, wherein the filter unit
A first body in which a blood canal through which blood can flow is formed;
a blood inlet provided at the front end of the blood canal to supply blood introduced from the outside to the blood canal;
A plasma canal disposed along the blood canal and formed in the body to be spaced apart from the blood canal; and
A plurality of cells are installed between the blood canal and the plasma canal along the longitudinal direction of the blood canal, and the plasma separated from the blood transported along the blood canal flows into the plasma canal, and the cross-sectional area through which the blood cells contained in the blood cannot pass A lab-on-a-chip for in situ analysis, characterized in that it comprises a plasma inflow channel formed to have. The method of claim 2, wherein the plasma canal
Lab for in situ analysis, characterized in that composed of a first plasma canal provided on one side of the blood canal centered on the blood canal, and a second plasma canal provided on the other side of the blood canal opposite to the one side centered on the blood canal on a chip. The method of claim 3, wherein the blood canal and plasma canal
A lab-on-a-chip for on-site analysis, characterized in that it is formed in a zigzag structure in the shape of 'L' or 'S'. The method of claim 1, wherein the plasma inlet plate
A lab-on-a-chip for on-site analysis, characterized in that a plurality of inlet grooves are formed in two or more lines on the surface in contact with the blood plasma passage. delete According to claim 1,
The lower substrate is a lab-on-a-chip for in situ analysis, characterized in that composed of a base plate formed of thermally conductive plastic or metal having a thermal conductivity of 15 to 400 W/mK, and a side plate formed in a vertical direction along an edge of the base plate. According to claim 2,
The lab-on-a-chip for in situ analysis, characterized in that the blood canal has a width of 30 to 60 μm and a depth of 10 to 20 μm, and the plasma canal has a width of 20 to 40 μm and a depth of 10 to 20 μm. According to claim 8,
The plasma inflow channel has a width of 4 to 6 μm and a depth of 2 to 3 μm. According to claim 1,
The upper substrate is provided with first intake holes penetrating in the vertical direction so that the air in the vacuum room can be easily discharged to the outside by shortening the moving path of the air stored in the vacuum room to form a vacuum state, and the plasma inlet plate Is formed to face the first intake hole and is provided with a second intake hole that penetrates in the vertical direction and communicates with the vacuum room,
The lab-on-a-chip for on-site analysis further comprises an intake hole sealing sheet sealing the first intake hole of the upper substrate so that the vacuum room maintains a vacuum state. According to claim 1,
The canal part is provided with a residual plasma inlet communicating with the end of the plasma passage so that residual plasma transported to the end of the plasma passage without moving to the inlet groove of the plasma inlet plate is discharged to the plasma inlet plate,
The lab-on-a-chip for on-site analysis, characterized in that, the plasma inlet plate is provided with a residual plasma discharge hole that faces the residual plasma inlet and communicates with the vacuum room to transfer the residual plasma discharged from the residual plasma inlet to the vacuum room. The method of claim 1, wherein the plasma inlet plate
A lab-on-a-chip for on-site analysis, characterized in that a target reagent is provided in the inlet groove.

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