WO2014030958A1

WO2014030958A1 - Method for preparing thin-film chip replica and microfluidic chip prepared by using the method

Info

Publication number: WO2014030958A1
Application number: PCT/KR2013/007570
Authority: WO
Inventors: 박상열; 김영호
Original assignee: 한국표준과학연구원
Priority date: 2012-08-24
Filing date: 2013-08-23
Publication date: 2014-02-27
Also published as: KR101410216B1; KR20140026075A

Abstract

The present invention relates to a method for preparing a thin-film chip replica and a method for preparing a microfluidic chip using the thin-film chip replica prepared by the method. More particularly, the present invention relates to a method for preparing a microfluidic chip comprising the processes of: preparing a thin-film chip replica through application, curing reaction, and releasing processes of a polymer solution for thin-film curing having a certain thickness by introducing the polymer solution for curing into an upper part of a micro fluidic chip mold and using a blade having microcrevices; and attaching a plane substrate on an upper part and a bottom part of the thin-film chip replica thus prepared. The micro fluidic chip having the thin-film chip replica according to the present invention can be variously applied to the diagnosis of diseases, food analysis and biochemical fields by analyzing specific amino acids, proteins, and so forth. Also, medical or biological experimental manipulation can be performed within the micro fluidic chip through tissue cell culturing and the cultured tissue cells.

Description

Specification

NAME OF THE INVENTION: A method for manufacturing a thin film chip replica and a microfluidic chip manufactured using the same

Field of technology

[1] The present invention relates to a method for manufacturing a thin film chip replica and a microfluidic chip using the thin film chip replica produced by the method.

Background

[2] Recently, micro-fluidic chips have been actively developed for use in ultra-fast disease diagnosis and high-efficiency bio-analysis. In a typical laboratory environment, a large amount of sample solution and

Lab-on chip (lab-on), which is carried out in the concept that various kinds of experiments performed using a semi-aqueous solution are carried out using chips smaller than a small slide glass size and having various structures of micrometer size inside Active development of a-chip shows the wide range of applications and future possibilities of microfluidic chips. In addition, active research and development in the field of micro-total analytical systems (TAS) using micro-Fulhui Dick chips can be used to quickly and efficiently analyze various types of biochemicals through miniaturized analysis chips.

[3] Microfluidic chips can be used for the analysis of chemical samples or biochemical samples, and can be used to culture tissue cells in microchambers and drug tests using cultured tissue cells. It can also be used for the mixing of two or more solutions, the synthesis by chemical reactions, and the production of functional materials. In particular, the volume of solutions in the microchannels and microchambers, the main constituents of the microfluidic chip, is extremely small, at levels below the nanoliters, and therefore when performing analysis using highly toxic solutions or substances within a single cell. In the case of very small sample volume, as in the analysis, the manipulation and analysis using the micro pulluidic chip is very useful. In addition, the microfluidic chip can be environmentally friendly due to the small amount of sample solution and semi-aqueous solution required and the amount of waste solution discharged.

[4] Silicon microprocessors are generally a method for producing micropuluidic chips.

It is a method of manufacturing by forming a micrometer-sized structure through an etching process on a hard ceramic material substrate such as a wafer or a glass plate and attaching it by covering and covering a flat substrate, or a flexible material.

Using a polymer material such as polydimethylene siloxane (PDMS) and the like, a method of manufacturing by soft-lithography process or the like is used. Regarding a method of manufacturing a thin film chip repullica or a microfluidic chip using the glass plate, the present invention is disclosed in Korean Patent No. 10-0788458 and the like. However, in the case of a micropoid chip manufactured using a silicon wafer or a glass plate, which is a hard material, adhesion between the microstructured plate and the flat plate is not only difficult to occur, but also a slight change in the manufacturing process and the usage is caused. It is easy to leak and has a weak point in the outer layer because of its ceramic material.

[5] In addition, Korean Patent Publication No. 10-2011-0038201 discloses a method for producing a microfluidic body having a micro structure thicker than several millimeters using a weak silicone rubber material such as polydimethylsiloxane (PDMS). have. However, in this case, the deformation of the microstructure and the connection of each component of the micropoid chip by the physical force applied from the outside, the pressure of the internal fluid generated from the internal microstructure, such as the microchannel or the micro-lamber, or the pressure of the gas, etc. Desorption and destruction at the edge are easy to occur.

[6] (prior art literature)

[Patent Document 1] Republic of Korea Patent Registration 10-0788458 (December 24, 2007)

[8] (Patent Document ¾ Republic of Korea Patent Publication 10- ^2011-00 ³⁸ ²⁰¹ ⁽²⁰¹¹ years (M month M-yl)

Detailed description of the invention

Technical task

[9] In order to solve the above problems, the present invention provides a method of manufacturing the thickness of a thin film chip replica, which is a main component of a micropuluidic chip, in a micrometer thin film. solid

Microfluidic chips with fixed, fixed plastic substrates are available. This makes it possible to manufacture micropoluidic chips that are more durable, can be manufactured in various sizes and shapes, and are more resistant to outer layers and physical forces than micropuluidic chips made of either flexible polymer materials or hard ceramic materials. .

Task solution

[10] In order to achieve this purpose, the present invention relates to a method of manufacturing a thin-film chip replica that can be manufactured to a certain thickness and to a microfluidic chip using the same.

[11] One aspect of the present invention,

[12] a) Sample solution inlet pattern _; Banung solution inlet pattern, micro channel pattern,

Applying the curable polymer solution to a microfluidic chip mold including a microchamber pattern and a solution outlet pattern;

B) hardening the curable polymer solution, the microfluidic chip

Releasing replica from the mold;

C) forming holes in the sample inlet, the reaction solution inlet and the solution outlet in the replica;

[15] relates to a method for manufacturing a thin film chip replica comprising the same.

[16] Another aspect of the present invention provides a method for bonding a flat substrate to the replica of step c). It relates to a thin film chip replica manufacturing method further comprising the step.

[17] Another aspect of the present invention relates to a microfluidic chip comprising the thin film chip-leple-lica, which is manufactured by the above method, wherein the microfluidic chip is formed on top of the thin film chip replica. ， The invention relates to a microfluidic chip which is a prefabricated microfluidic chip further comprising a lower part or a fixing part at a top and a bottom part thereof.

[18] Another aspect of the present invention,

[19] an upper fixing member including a fixing groove for connecting the tube and a male injection tube;

A thin film chip replica positioned below the upper fixing part;

[21] The shape of the thin film chip replica is disposed under the thin film chip replica.

A flat substrate to hold;

A lower fixing member disposed below the planar engine and including a screw groove;

[23] a male screw inserted into the male screw injection tube of the upper fixing member and fixed by the screw groove of the lower fixing member;

The invention relates to a microfluidic chip containing [24].

[25] A method of manufacturing a thin film chip replica proposed by the present invention and using the same

The micro pulluid chip will be described in detail.

[26] The first step a) is a sample solution inlet pattern, a semi-aqueous solution inlet pattern ，

By using a curable polymer solution in a microfluidic chip mold including a microchannel pattern, a microchamber pattern and a solution outlet pattern

Proceed with replication (plastic repetition).

[27] The microfluidic chip mold used in the present invention is a microsystem

Devices should be manufactured using materials that can be easily mass-produced using processes such as injection molding, hot embossing, casting, and laser micromachining. Preferably polyamide (PA),

'Polybutylene terephtalate (PBT),

Polycarbonate (PC), polyethylene (PE, PE),

Polymethylmethacrylate (PMMA),

Polyoxymethylene (POM), polypropylene (PP), polyphenylenether (PPE), polystyrene (PS), polysulfone (polysulfone, PSU), liquid crystal polymer (liquid crystal polymer) , LCP),

Polyetheretherketone (PEEK),

Polyetherimide (PEI), polylactide,

Polydimethylsiloxane (PDMS), cycloolefin

Cycloolefm copolymer (COC), polyethylene glycol

It is advisable to use polyethyleneglycol diacrylate.

[28] The microfluidic chip mold used in the present invention is subjected to a micromachining process for performing operations such as sawing, cutting, and milling using micro equipment, or silicon micromachining and lithography using silicon itself. By all means It may be prepared, preferably using an etching process. In the etching process, a mold may be manufactured by depositing a thin film such as oxide or nitride on the silicon, coating a photoresist, and irradiating UV light to obtain a desired pattern.

[29] The curable polymer solution used in the present invention may contain a polymer resin and a curing initiator.

It can be prepared by mixing. It is preferable to use a silicone polymer or an acrylic polymer as the curable polymer solution. The silicone polymer may be formed of a polysiloxane, a polysilane, a polycarbosilane, a polysilazane, or the like to form a main chain, and the side chain may include hydrogen, ethyl, Propyl, butyl, phenyl, and the like, preferably

It is preferable to use polydimethylsiloxane (PDMS). In addition, the acrylic polymer is preferably an acrylate (acrylate) or

It is preferable to use an acrylic polymer having a functional group of methacrylate.

[30] The polymerization initiator may be in any kind within the scope of not impairing the object of the present invention.

It is not limited and it is preferable to use a photocuring initiator or a thermal polymerization initiator.

The photocuring initiator may be added to be used with an acrylic polymer precursor (prepolymer). Preferably

2-hydroxy-2-methylpropiophenone,

2,6-di- (P-azidobenzal) -4-methylcyclonucleanone (2,6-di- (p-azidobenzal) -4-methylcyclohe xanone), and

2,6-di- (p-azidobenzal) -cyclonucanonone (2,6-di- (p-azidobenzal) -4-cyclolie) ianone) bisazide compound, benzophenone,

Methyl benzoylbenzoate, methyl benzoylbenzoate,

4,4-bis (dimethylaminobenzophenone) (4,4'-bis (dimethylaminobenzophenon)),

4,4-dichlorobenzophenon,

4-benzoyl-4'-methyldiphenylketone (4-benzoyl-4'-methyldiphenylk: etone),

Dibenzylketone, 2,2'-diethoxyacetophenone, (2,2'-diethoxy

acetophenone),

1-phenyl-1, ^{^{2-butanedione-2-}} (o- ethoxy carbonyl knife on) oxime ^{(I -phenyl-butandione- 2 - (} o-ethoxycarb onyl) oxime),

1,3-diphenyl-propanedi-2- (o-ethoxycarbonyl) oxime (l, 3-diphenyl-propandione-2- (eth oxycarbonyl) oxime),

1-phenyl-3-ethoxy-propanedione-2- (o-benzyl) oxime (l-phenyl-3-ethoxy-propandione-2- (ob enzyl) oxime), N-phenylglycidyl ),

3-phenyl-5-isoxazolone (3-phenyl-5-isoxazolone),

1-hydroxycyclohexylphenylke (I-hy droxycy clohexy lpheny lketone), 2-methyl- (4- (methylthio) phenyl) -2-morpholino-1-propanone (2-methyl- (4-methykhio) phenyl-2-morpholino-L-propanone),

Naphthalenesulfonylchloride,

Quinolinessulonylchloride,

N-phenylthioacridone,.

4,4'-azobisisobutyronitile,

Diphenyl disulfide (diphenyl sulfide),

Benzthiazole disulfide,

Triphenylphosphine, camphorquinone,

Carbontetrabromide,

Tribromophenylsulfone, benzoylperoxide, dimethylmethylhydrogensiloxane,

Dimethoxy phenyl acetophenone,

Dicumylperoxide and the like can be used.

The thermosetting initiator is preferably an α-aminoalkylphenone compound, an acylphosphine oxide compound, an oxime ester compound, a benzophenone compound having an amino group or a benzoic acid ester compound having an amino group. Specifically

2-methyl- [4- (methylthio) phenyl] -2-morpholinopropane-1-one (2-methyl- [4- (methylthio) phenyl

1- 2-morpholinopropane-l-one),

2-Dimethylamino-2- (4-methylbenzyl) -1- (4-morpholin-4-yl-phenyl) -butan-1-one (2-dimethylamin

0- 2- (4-methylbenzyl) -1-(4-morpholine-4-yl-phenyl) -butane-1 -one),

2,4,6-trimethylbenzoylphenylphosphine oxide,

Bis (2,4,6-trimethylbenzoyl) -phenylphosphineoxide (bis (2,4,6-trimethylbenzoyl) -phen ylphosphineoxide),

Bis (2,6-dimethoxy benzoyl)-(2,4,4-trimethylpentyl) -phosphine oxide (bis (2,6-dimetho xybenzoyl)-(2,4,4-trimethylphentyl) -phosphineoxide),

1-phenyl-1,2-propanedione-2- (ethoxycarbonyl) oxime (I.-phenyl-1,2-propanedione-2- (o -ethoxycarbonyl) oxime), 1,2-octanediol ( 1,2-octanediol),

1- [4- (phenylthio) -2- (o-benzoyloxime)] (1 [XphenyUhkO ^ o-benzoyloxime),

1-phenyl-1,2-butadione-2- (o-methoxycarbonyl) oxime (l-phenyl-l, ² -butadione- ^2- (o-meth oxycarbonyl) oxime),

1,3-diphenylpropanetrione-2- (o-ethoxycarbonyl) oxime (l, 3-diphenylpropanetrion- ^2- (o -ethoxycarbonyl), ethanone, l- (o-acetyloxime) (l- (o-acetyloxime)),

4,4-bis (dimethylamino) benzophenone (4,4-bis (dimethylamino) benzophenone), p-dimethylaminobenzoic acid ethyl,

2-ethylhexyl-p-aminobenzoate or the like may be used. The dye added to the curable polymer solution can be used irrespective of the type within the scope of not impairing the object of the present invention.

The curable polymer solution used in the present invention may be prepared by including 90 to 99.9% by weight of a polysiloxane silicone polymer or an acrylic polymer and 0.1 to 10% by weight of a photocuring initiator.

In the present invention, the plastic replication is not limited to the type within the scope of achieving the object of the present invention, preferably using any one of a casting method, a hot embossing method and an injection molding method. good. More preferably, it is preferable to use a casting method using a blade because the thickness of the thin film chip replica to be produced can be uniformly formed.

[36] The blade used in the casting method should have a certain size at the blade. The shape of the blade including the groove may be formed of a yaw (HI) to uniformly form the thickness of the thin film chip repolya, and the length of the groove may be freely added or reduced depending on the size of the thin film chip replica to be manufactured. It is possible to adjust the thickness of the thin film chip replica manufactured by adjusting the spacing between the blade and the micro pulluidic chip mold in the range of 100 to 3000.

[0037] The casting method preferably applies the curable polymer solution on top of the microfluidic chip mold, and then the micropuluidic chip mold is fixed and the blade moves in the uniaxial direction. After the blade is fixed, the microfluidic chip mold may move in a uniaxial direction to effectively apply the curable polymer solution on the microfluidic chip mold.

Next, in step b), the curable polymer solution is cured.

The cured replica is released from the microfluidic chip mold. remind

The curing reaction is recommended to proceed with the curing reaction by applying heat or ultraviolet rays. In the case of the thermosetting reaction, it is recommended to react for 10 minutes to 70 hours at a temperature of 10 to 200 ^o C. In the case of ultraviolet curing, it is preferable to proceed by irradiating ultraviolet rays having a wavelength in the range of 200 to 400 nm for 5 seconds to 30 minutes. If the temperature is less than 10 ^o C, the curing reaction is delayed, and if it exceeds 200 ° C, pyrolysis reaction by high heat occurs. If less than 10 minutes when the thermal curing reaction, the hardening does not occur sufficiently, if it exceeds 70 hours, cost problems due to long-term heat treatment will occur. When irradiating for less than 5 seconds during the self-curing, the curing does not occur hardly, and when irradiated for 30 minutes or more, photolysis reaction by ultraviolet irradiation occurs. The thin film chip replica of the curing reaction is preferably maintained in a constant thickness so that the manufactured microfluidic chip can perform the experiment and diagnostic function smoothly. Preferably, the thickness deviation of the thin film chip replica may be -5jum to 5 / / m. Here, the thickness deviation means the difference between the highest and lowest points of the cured thin film chip replica. (In thickness deviation It is written in the meaning.Please confirm.)

[39] When the curing reaction is finished, the cured resin is cured from the microfluidic chip mold.

Release the replica. If the thin-film chip replica is made of flexible silicone material, it is advisable to release it with caution since the deformation may occur in the thin-film chip replica.

[0040] The step c) is a sample inlet, a reaction solution inlet and the cured replica

And forming a hole in the solution discharge port.

The semi-aqueous solution inlet and the solution outlet may be engraved, and it is preferable to form an open hole in the upper and lower portions of the thin film chip replica. In this step, a metal or plastic tube having a diameter of 50 to 5000 m is used. The hole can be formed using a knife that is pressed or has a sharp blade.

In the present invention, after the step c) is completed, the thin film chip replica is applied.

Bonding the flat substrate; the flat substrate may be formed of a ceramic or polymer material on the plate, and preferably a flat substrate or acrylic based material, such as glass, quartz, silicon dioxide, etc. It is preferable to use a flat substrate composed of urethane, olefin, and silicon polymer materials.

[42] The thin film chip may be bonded to the planar substrate by using the thin film chip repolya.

A method of bonding by using a unique adhesive property of the replica or by performing mutual plasma and ultraviolet-ozone welding treatment on the mutual contact surface of the thin film chip replica and the planar substrate for 10 seconds to 40 minutes, then performing mutual contact or laminating, It is advisable to bond by either pressure, laser welding or ultrasonic welding.

[43] The thin film chip replica manufactured by the above manufacturing method is placed on the flat substrate.

Microfluidic chips can be used as they are, or they can withstand the external impact or physical forces desired by the present invention, including the upper, lower or upper and lower fixing frames of the thin film chip replicas. ，

[44] an upper fixing frame including a fixing groove and a male screw injection pipe for connecting a tube;

A thin film chip replica having a lower portion of the upper portion fixed thereto;

[46] The shape of the thin film chip replica is placed under the thin film chip replica.

Retaining flat substrates;

A lower fixing member disposed below the flat substrate and including a screw groove;

A male screw injected into the male screw injection tube of the upper fixing member and fixed by the screw groove of the lower fixing frame;

Prefabricated microfluidic chips containing [49] are preferred.

[50] In this case, if one or more of the components of the microfluidic chip including the thin film chip replica are required to be replaced, the assembled micropuluidic chip may be dismantled by manipulating the male screw and replaced with a new component. By assembling, it is possible to reassemble and reuse.

[51] Fixing grooves for connecting feed-injection tubes connected to feed-injection tubes fixed to upper and upper parts, fixing grooves for connecting reaction solution injecting tubes to which the reaction solution injection valves are connected;

The solution discharge tube is connected to the solution discharge tube connection groove, it may be made of a structure having a plurality of male screw injection tube. The upper fixing frame is preferably located on the top of the thin film chip replica bonded to the flat substrate, a transparent material that transmits most of the light in the visible light in the 400 to 800nm range, or the optical signal detection portion is a transparent material, It is preferable that the part of is made of an opaque material, and preferably made of a resin composition composed of transparent, opaque or a combination thereof.

[52] The resin composition is preferably polyamide, polyacryl,

One or two or more of a polymer resin such as polymethyl methacrylate, epoxy, polystyrene, polyoxy methylene, polyethylene, polypropylene, and polycarbonate, or one or more of the polymer resin as a polymer composite material It is preferable to use those prepared by mixing silica-based inorganic particles such as carbon black, silica, and silicon dioxide in two or more.

[53] a fixing groove for connecting the sample injection tube, a fixing groove for connecting a positive injection tube, and

Fixing groove for connecting the solution discharge tube is to function to connect and secure each tube connecting part connected from the top, it is easy to connect with the female thread acid on the inner wall, and the upper fixing is configured to penetrate the upper and lower From each of the rib connection portion disposed in the tester to facilitate the function to easily transfer the desired semi-aqueous solution or solution to the thin film chip replica.

The lower fixing is made of the same or different polymer resin as the upper fixing frame.

It may be prepared, preferably the resin composition, the polymer composite material, or a mixture thereof, or silica-based, such as quartz, glass, silicon dioxide, etc.

It is better to use an inorganic material.

In addition, the lower fixing frame may have a screw groove and a flat substrate seating groove. The screw groove receives the male screw passing through the upper fixing to serve to firmly fix the upper fixing frame and the lower fixing frame, the flat substrate seating groove is formed as an intaglio groove on the upper surface of the lower fixing frame of the flat substrate You can set the location.

The male screw may be made of a material including a metal or a polymer resin, and after the upper fixing is injected into the male screw injection tube,

The lower fixture is injected into and fixed in the screw groove of the prefabricated microfluidic chip so that it can be used for assembly or separation.

Effects of the Invention

A thin film chip replica manufactured by the method for manufacturing a thin film chip replica according to the present invention; The microfluidic chip using this is very simple to manufacture, and it is a term for the thickness-adjustment of the thin film chale plaques to be manufactured: It is also possible to fix the upper, lower or upper part and the lower part of the thin film chip replica to the flat substrate. And by installing a solution injection or discharge tube, the sample or solution can be easily introduced into the microfluidic chip, eliminating the problem of structural deformation in physical forces or impacts.The manufacturing method is simple and does not require expensive equipment. They can be mass-produced at a low cost in a typical workspace, and can be assembled and separated, making it easy to combine and replace components and to manufacture them according to their intended use.

Brief description of the drawings

1 and 2 illustrate a method of manufacturing a thin film chip replelica according to one embodiment of the present invention.

It is shown schematically.

3 shows the assembly form of the components of the micropuluidic chip according to the present invention.

It is a schematic diagram showing.

4 shows a perspective view in which the components of the microfluidic chip according to the present invention are assembled.

5 shows a production and an embodiment of a thin film chip replica manufactured according to the present invention.

6 shows an experiment using a microfluidic chip manufactured according to the present invention.

7 shows a photograph of a microfluidic chip according to the present invention.

FIG. 8 is a photograph showing an embodiment using a micropuluidic chip.

9 is a photograph showing an embodiment of manufacturing a microfluidic chip including an upper and a lower fixing frame according to the present invention.

[66] (symbol description)

[67] 10 microfluidic chip molds

[68] 11 microfluidic chip pattern

[69] 12 Sample solution inlet and pattern

[70] 13 Banung solution inlet pattern

[71] 14 microchannel patterns

[72] 15 Microchamber Patterns

[73] 16 outlet pattern

[74] 20 Y-shaped blade

[75] 21 blade microgrooves

[76] 30 Curable Polymer Solution

[77] 31 curable polymer thin film

[78] 40 thin film chip replicas

[79] 41 microchambers [80] 42 sample inlet

[81] 43 -Bungungyong ^Pakju ᅵ ^Apgu

[82] 44 microchannels

[83] 47 solution outlet

[84] 50 flat substrates

[85] 60 upper fixing frame

[86] 61 fixing grooves for connection of sample injection tubes

[87] 62 Locking grooves for connecting semi-aqueous solution injection tubes

[88] 63 fixing groove for connection of solution discharge tube

[89] 65 male thread injection pipe

[90] 70 Lower fixing

[91] 71 thread groove

[92] 72 flat substrate seating grooves

[93] 81 Sample injection tube connection

[94] 82 Reaction solution injection tube connection

[95] 83 Solution outlet tube connection

[96] 85 Sample Injection Tubes

[97] 86 Reaction Solution Injection Tube

[98] 87 solution discharge tubes

[99] 89 male thread

[100] .100: Assembly Micro Pulley Tooth Chip

Best Mode for Carrying Out the Invention

Hereinafter, the microfluidic chip manufactured using the thin film chip replica manufacturing method and the invention of the present invention will be described in more detail with reference to the accompanying drawings and examples. The descriptions and examples in these drawings are only for explaining the present invention in more detail, and the scope of the present invention is not limited to these descriptions and examples. In addition, the following terms are terms defined in consideration of the functions of the present invention, which may vary according to the intention or the custom of the user or the operator. Therefore, the definitions of these terms should be based on the contents throughout the specification. ᅳ

[102] process for manufacturing thin film etched polypropylene

1 and 2 illustrate a thin film chip replica manufacturing process according to one embodiment of the present invention.

1 (a) shows a sample solution inlet pattern 12, a semi-aqueous solution inlet pattern 13, a micro channel pattern 14, a micro chamber pattern 15, and a solution outlet pattern 16. As shown in FIG. A microfluidic chip mold (10) having a microfluidic chip pattern (11) formed with a rounded relief is shown. When the mold is prepared, it has a blade fine groove (21) as shown in (b) (four) shaped blade (20). Micro Place on top of the fluidic chip mold. At this time, the lower end of the groove of the blade having the fine groove and the upper surface of the micro pulley Dick needle 10 is maintained to have a constant interval.

After the placement on the blade is completed, as shown in (c) of the microfluidic chip mold

The curable polymer solution 30 is injected close to the blade having the fine gap. And as (d), (e) the blade is applied to the top of the microfluidic chip mold as ¾ when moving from one end to the opposite end of the microfluidic chip mold. The thin film thickness of the curable polymer solution applied at this time is determined by the distance between the lower end of the center portion of the blade microgroove and the upper surface of the microfluidic chip mold, and the lower end of the blade microgroove and the micro pullupic chip mold. By adjusting the distance between the upper surfaces, a curable polymer thin film 31 of various thicknesses can be produced.

When the curable polymer thin film is manufactured, the curable polymer as shown in FIG.

The thin film 31 is thermally cured for a predetermined time or irradiated with ultraviolet rays to cause the photocuring reaction. At this time, the cured polymer thin film 31 is transformed into semi-solid and solid forms according to the degree of curing reaction, thereby facilitating the mold release in the next step. FIG. 2G is a view of releasing the thin film chip repulica 40 formed by curing the curable polymer thin film from the micro pulley dick chip mold 10.

(H) is a thin film chip replica completely released from the microfluidic chip mold to obtain a thin film chip replica 40 in which structures such as the microchamber 41 are intaglio.

[107] After the mold is finished, the sample inlet 42 of the thin film chip replica 40, as shown in (i),

A hole is formed in the reaction solution inlet 43 and the solution outlet 47 to finally prepare the thin film chip replica having the microchannel 44 and the microchamber 41.

[108] Prefabricated Micro Polui ¾ ¾ containing Thin Film Saliva Replicarom

3 to 4 are structured on the upper and lower portions of the thin film chip replica of the present invention.

The manufacturing process of the prefabricated micro pulley dick chip with the fixing frame is shown.

3 illustrates each component of the prefabricated microfluidic chip 100 and a state before assembly of the component. The fixing groove for connecting the tube is composed of the fixing groove (61) for connecting the sample injection stream, the fixing groove (62) for connecting the reaction solution injection tube, and the fixing groove (63) for connecting the solution discharge tube. A thin film chip replica 40 disposed on the lower portion of the upper substrate 60 having the upper fixing 65 thereon, a flat substrate 50 disposed on the lower portion of the thin film chip replica, and a lower portion of the flat substrate 50. The lower fixing 70 is disposed in the screw groove 71 and the flat substrate seating groove 72, the male screw 89 is injected into the male screw injection pipe and the lower screw is fixed to the screw groove of the lower screw assembly. [1 1 1] The upper fixing frame is a fixing groove (61) for connecting the sample injection tube, a fixing groove (62) for connecting the semi-aqueous solution injection stream, a fixing groove (63) for connecting the solution discharge tube, and a plurality of male thread injection tubes (65) In addition, the upper fixing (60) is made of a transparent and rigid plastic material that transmits most of the light in the visible light region, so that the inside of the microchannel 44 or the microchamber 41 of the microfluidic chip is made. It can be easily observed and can easily detect fluorescence or color change during sample analysis.

[112] Fixing groove 61 for connecting the sample injection flow tube, for connecting the reaction solution injection tube

The fixing groove 62 and the fixing groove 63 for connecting the solution discharge tube serve to connect and fix the respective tube connecting portions 81, 82, and 83 connected from the upper side, and the female thread is connected at the inner wall. This easy and vertically penetrating structure functions to transfer the solution from the respective tube connection portions 81, 82, and 83 arranged at the top to the bottom.

The lower fixing (70) is a flat solid ceramic or plastic or metal

It is made of a material, and has the screw groove (7.1) having a screw groove 71 on the inner wall so that the male screw (89) is injected and fixed.

The flat substrate seating groove 72 is formed in the form of an intaglio groove on the upper surface of the bottom fixing 70, and positions the flat substrate 50 to be seated at a specific position when the flat substrate 50 is placed. Plays a role.

The male screw 89 is made of metal or plastic material.

After the upper fixing is injected into the male screw injection tube (65) of (60)

The lower fixture is injected into the screw groove 71 of the 70 to be fixed and used to assemble or detach the prefabricated microfluidic chip 100.

4 is a perspective view showing a state in which the components shown in FIG. 3 are assembled. When one or more components of the prefabricated microfluidic chip 100 having the thin film chip replica of the present invention are required to be replaced, the assembled micropuluidic chip is dismantled by manipulating the male screw 89 and a new construction is performed. It can be reassembled and reused by injecting the components and assembling them.

Hereinafter, a thin film chip repolya prepared by the method according to the present invention and the same

A process of manufacturing a microfluidic chip including will be described in detail through an embodiment. However, the following examples are only one example to assist the preferred embodiment of the present invention, the present invention is not limited to the examples.

[118] (Example 1)

[1 19] A microfluidic chip was manufactured in order to confirm the manufacturing process of the thin film chip replica manufactured by the above method and the function of the micro pulluidic chip having the thin film chip replica.

[120] First, microfluidic chip mold is polyethylene glycol

Diacrylate (polyethyleneglycoldiacrylate, average molecular weight 258, Sigma-Aldrich) 96.9 weight% solution, photocuring initiator

2-hydroxy-2-methoxy, naphthyl ⁱ profile non Opaque (2-hydroxy-2-methylpiOpiopheiK ) ne, Sigma-Aldrich , Inc.) 3% by weight, and a red dye (Rhodamin B, Sigma-Aldrich, Inc.) 0.1% by weight Was mixed to prepare a UV curable polymer solution for a microfluidic chip mold.

A small amount of red dye added at this time was used for the purpose of simply coloring to facilitate the observation of the structure of the micro pulley dick chip mold to be produced.

[122] A photomask of plastic film material manufactured by Microtech Co., Ltd. is coated on the glass plate with a thickness of 150 μl of the UV-curable polymer solution for the microfluidic chip mold and has a transparent window in the shape of a microfluidic chip. And irradiate with ultraviolet rays emitted from an ultraviolet lamp having a maximum self-emission wavelength of 360 nm and emitting ultraviolet rays in the range of 200 to 400 n at the top for 10 seconds, to the microfluidic chip-shaped portion of the photomask only. Ultraviolet rays were irradiated to allow the UV-curable polymer solution for microfluidic chip mold disposed below to be selectively cured and solidified. Then, the photomask was removed, and the UV curable polymer solution for the microfluidic chip mold of the portion not irradiated with UV was removed, and finally, a microfluidic chirp mold having an embossed microfluidic chip-like structure was prepared. The photo of the micro pulley dick chip mold thus prepared is shown in Fig. 5A.

A polydimethylsiloxane (PDMS) polymer precursor solution (trade name: Sylgard 184 A, Dow) on top of a micro pulluid chip mold having a height of 150 / mi embossed microfluidic chip-shaped structure prepared as described above. Corning Co., Ltd.) 93% by weight and the curing agent solution (trade name: Sylgard 184 B, Dow Corning Co., Ltd.) 7% by weight of a mixture of thin film to 320 thickness using a blade having a groove depth of 320 / ΛΠ and a length of 6cm Curing reaction was performed for 15 minutes at 85 ° C on a hotplate hotplate. Then, the polydimethylsiloxane thin film chip replica cured as shown in Fig. 5A is lifted from one end of the microfluidic chip mold.

Release was performed. After the release process, the sample inlet, the reaction solution inlet and

The thin film chip replica of the present invention finally prepared by punching a hole in the solution outlet portion is shown in FIG. At this time, the thickness of the polydimethylsiloxane thin film chip replica manufactured was 320 ^, and in particular, the upper thicknesses of the microchannel and microchamber portions were 170 zm. The thin film chip replica prepared as described above was attached to a glass flat substrate using the inherent tacky niol of the polydimethylsiloxane thin film chip replica to prepare a microfluidic chip having a basic form as shown in FIG.

[124] A drop of red ink solution is dropped into the sample inlet and the reaction solution inlet in order to confirm the operation of the micro pulluidic chip having the manufactured thin film chip replica. Dropping was confirmed the moving direction of the ink is shown in FIG.

4 and (b) show that a drop of red and ink solution was dropped one by one at the sample inlet and the reaction liquid inlet. Subsequently, as shown in (c), the capillary phenomenon due to the hydrophilicity of the surface of the flat substrate showed that the red ink was transferred through the microchannel from the sample inlet and the semi-aqueous solution inlet. After 10 seconds, the red ink reached the solution outlet through the microchannel and the microchamber, which is shown through (D) of FIG. As in the above embodiment, it was confirmed that the microfluidic chip manufactured by the method of the present invention had a dividing function.

[126] (Example 2)

In order to verify the function of the prefabricated micro pulluidic chip by the method of the present invention, a micro pulluidic chip was manufactured. First, a method and a process of manufacturing a thin film chip replica in a prefabricated micro pulluidic chip were manufactured in the same manner as in Example 1, and the thin film chip replica was bonded to a flat substrate on the bottom.

In order to install the fixing frame on the upper and lower portions of the thin film chip repolia, milling was performed using an acrylic transparent plastic plate to manufacture upper and lower fixing parts. Thereafter, the upper fixing was injected into the male screw injection tube using the male screw, thereby fixing the upper fixing and the lower fixing to manufacture the assembled micro pulluidic chip. 7 and 9 show that the sample injection tube and the solution discharge tube are connected to the manufactured prefabricated microfluidic chip.

[129] The fabricated prefabricated microfluidic chip used the same red ink as Example 1.

Injecting a red ink through the sample injection tube using the ink was confirmed the direction of movement of the ink is shown in FIG.

[130] The red ink injected through the sample injection tube as shown in (b) of FIG.

It was confirmed that the transfer to the microchannel and the microchamber. This of Figure 8

It is shown in (C) and (D). A process in which red ink is injected and transferred to a microchannel and a microchamber that the prefabricated microfluidic chip having the thin film chip replica of the present invention as in Example 2 has a sufficient function as a general micropuluidic chip. I could confirm through. In particular, as the upper plastic substrate is hard and transparent, the color of the prefabricated microfluidic chip having the thin film chip replica of the present invention can be observed in the process of analyzing a specific analyte as shown in the clear color of the red ink. It was possible to detect by fluorescence emission.

[131] Although the preferred embodiment of the present invention has been described above, the present invention

It is not limited to the above specific embodiment. That is, those skilled in the art to which the present invention pertains can make many changes and modifications to the present invention without departing from the spirit and scope of the appended claims, and all such appropriate changes and modifications Equivalents should be considered to be within the scope of the present invention.

Claims

Claim

[Claim 1] a) a sample solution injection hole patterns, the reaction solution inlet patterns, micro pool Louis dikchip ^"replication step with the curable polymer solution into a mold comprising a micro-channel pattern, micro chamber pattern and a solution outlet pattern;

b) curing the curable polymer solution to release a replica from the micropuluid chip mold;

c) forming holes in the sample inlet, the reaction solution inlet and the solution outlet;

Thin film chip replica manufacturing method comprising a.

[Claim 2] In paragraph 1,

and d) adhering a planar substrate to the replica of step c).

[Claim 3] In paragraph 1,

The method of manufacturing a thin film chip replica, wherein the replication is any one of a casting method, a hot embossing method, and an injection molding method.

[Claim 4] In paragraph 3,

The repetition. The casting method uses a casting method, wherein the casting method adjusts the distance between the blade and the micropuluidic chip mold to 100 to 3000.

[Claim 5] In paragraph 1.

The curable polymer solution is a polysiloxane-based polymer or acrylic polymer comprising 90 to 99.9% by weight and a curing initiator 0.1 to 10% by weight of a thin film chip replica manufacturing method.

[Claim 6] In paragraph 1,

The hardening reaction is heat curing or ultraviolet curing, thin film chip replica manufacturing method.

[Claim 7] Regarding Section I,

The c) step is to form a hole in which the upper and lower portions of the replica open.

[Claim 8] In paragraph 1,

The thickness deviation of the cured replica is -5 to +5 thin film chip replica manufacturing method.

Claim 9 A micropoid chip comprising a thin film chip replica manufactured by the method of any one of paragraphs 1 to 8.

[Claim 10] In the paragraph ⁹ ,

The microfluidic chip is formed on top and bottom of the thin film needle replica Or prefabricated micros further comprising top and bottom fastening

Fluoric-Picked Pyrofluidic Chip.

[Claim 11] In paragraph 10,

An upper fixation including a fixing groove for the tube connection and a male injection tube; A thin film chip replier positioned below the upper fixture;

A planar substrate disposed under the thin film chip replica to maintain a shape of the thin film chip replica;

A lower fixing frame disposed below the flat substrate and including a screw groove; A male screw injected into the male screw injection tube of the upper fixing member and fixed by the screw groove of the lower fixing member;

Micropuluid chip containing.

[Claim 12] In paragraph 11,

The fixing groove for connecting the tube includes a fixing groove for connecting the sample injection tube to which the sample injection tube is connected, a fixing groove for connecting the reaction liquid injection tube to which the reaction solution injection tube is connected, and a solution discharge flow connection connection groove to which the solution discharge tube is connected. 2D chip.

13. The method of claim 12,

The fixing groove for connecting the sample injection tube, the fixing groove for connecting the reaction solution injection tube, and the fixing groove for connecting the solution discharge tube have a female thread on the inner wall, and the fixing grooves have the upper fixing frame perpendicular to the upper surface of the upper fixing frame. Microfluidic chip having a structure that penetrates through.

[Claim 14] The clause of claim 11,

The lower fixing microfluidic chip comprising a screw groove and a flat substrate seating groove, wherein the screw groove is injected by the male screw passing through the male screw injection pipe of the upper fixing frame.

[Claim 15] In paragraph 13,

The flat substrate seating groove is a microfluidic chip which is formed in the upper surface of the bottom fixing grooves of the intaglio to position the flat substrate.

[Claim 16] In paragraph U,

And wherein the male screw comprises a metal or a polymer resin, wherein the lower fixing is injected into the screw groove of the male screw injection tube to fix the lower screw.