KR101716302B1 - Manufacturing method of biochemical reactors - Google Patents

Manufacturing method of biochemical reactors Download PDF

Info

Publication number
KR101716302B1
KR101716302B1 KR1020150147028A KR20150147028A KR101716302B1 KR 101716302 B1 KR101716302 B1 KR 101716302B1 KR 1020150147028 A KR1020150147028 A KR 1020150147028A KR 20150147028 A KR20150147028 A KR 20150147028A KR 101716302 B1 KR101716302 B1 KR 101716302B1
Authority
KR
South Korea
Prior art keywords
mold
flow path
photosensitive material
nano
microchannel
Prior art date
Application number
KR1020150147028A
Other languages
Korean (ko)
Inventor
김태성
김민석
Original Assignee
울산과학기술원
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 울산과학기술원 filed Critical 울산과학기술원
Priority to KR1020150147028A priority Critical patent/KR101716302B1/en
Priority to PCT/KR2016/005639 priority patent/WO2017069364A1/en
Application granted granted Critical
Publication of KR101716302B1 publication Critical patent/KR101716302B1/en

Links

Images

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03F—PHOTOMECHANICAL PRODUCTION OF TEXTURED OR PATTERNED SURFACES, e.g. FOR PRINTING, FOR PROCESSING OF SEMICONDUCTOR DEVICES; MATERIALS THEREFOR; ORIGINALS THEREFOR; APPARATUS SPECIALLY ADAPTED THEREFOR
    • G03F7/00—Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/12—Specific details about manufacturing devices

Abstract

The present invention relates to a biochemical reactor and a method of manufacturing the same, comprising: a substrate; And a first flow path formed on the substrate and capable of supplying and flowing a fluid including microorganisms, wherein the first flow path is communicated with the first flow path, and the movement of the microorganisms among the fluid flowing through the first flow path A first microchannel loading unit having at least one first integrated space formed to guide the microchannels to the microchannels and integrating the microchips, and a first induction flow channel connecting the first flowpath and the first integrated space; And a second flow path which is separated from the microchannel loading part and through which a fluid containing a bio material for gene expression of the microorganisms can be supplied and flows, A second accumulation space through which the biomaterial can be applied and a second accumulation space through which the second flow path and the second accumulation space are connected, The micro / nano fluidic channel block includes a second microchannel loading part having a channel formed therein and a nano channel part connecting the first accumulation space and the second accumulation space to supply the biomaterial to the microorganisms. .

Description

[0002] Manufacturing methods of biochemical reactors [0003]

The present invention relates to a biochemical reactor and a method of manufacturing the same. More particularly, the present invention relates to a biochemical reactor and a biochemical reactor which are manufactured using micro- and nano-sized microchannels formed through a photolithography process, Reactor and a method for producing the same.

Generally, microfluidic devices have been used for the analysis of microorganisms, in which the diffusion is controlled by a nanopore membrane or a hydrogel, but the convection is prevented. For example, a nanopore membrane made of polyethylene or polycarbonate was sandwiched between microchannels made of polydimethylsiloxane (PDMS) and used as a diffusion layer to minimize convective migration of microorganisms.

The microfluidic devices having such a nanopore membrane can prevent unnecessary convective flow while allowing necessary diffusion. However, there is a problem that it is difficult to observe the microfluidic device due to the fluid leakage or the opacity of the nanoporous membrane. In addition, when long-term experiments are performed, there is a problem that the nanopore film is deformed or decomposed to degrade the accuracy of the experiment.

In recent years, in addition to the conventional photolithography, soft lithography, electron beam lithography (e-beam), and the like have been developed in addition to the development of micro- and nanofluidics lithography, nanoimprint lithography, and the like are being developed.

Korean Patent No. 10-1206619

The present invention aims to provide a biochemical reactor which is fabricated by using micro- and nano-sized microchannels formed through a photolithography process and can test gene expression of various kinds of microorganisms and a method for manufacturing the same.

The present invention provides a semiconductor device comprising: a substrate; And a first flow path formed on the substrate and capable of supplying and flowing a fluid including microorganisms, wherein the first flow path is communicated with the first flow path, and the movement of the microorganisms among the fluid flowing through the first flow path A first microchannel loading unit having at least one first integrated space formed to guide the microchannels to the microchannels and integrating the microchips, and a first induction flow channel connecting the first flowpath and the first integrated space; And a second flow path which is separated from the microchannel loading part and through which a fluid containing a bio material for gene expression of the microorganisms can be supplied and flows, A second accumulation space through which the biomaterial can be applied and a second accumulation space through which the second flow path and the second accumulation space are connected, The micro / nano fluidic channel block includes a second microchannel loading part having a channel formed therein and a nano channel part connecting the first accumulation space and the second accumulation space to supply the biomaterial to the microorganisms. Wherein the first microchannel loading portion, the second microchannel loading portion, and the nanochannel portion side of the microchannel / nanofluid channel block are disposed toward the substrate, and the microchannel / A block is adhered to a biochemical reactor.

According to another aspect of the present invention, there is provided a microfluidic system including a first flow path through which a fluid containing microorganisms flows and flows, a first integrated space in which the microbes are integrated, A first protruding structure having a shape corresponding to a first microchannel loading part formed with a first induction channel connecting the flow path and the first integrated space is formed and spaced apart from the first protruding structure, A second flow path through which the fluid is supplied and flows, a second accumulation space communicating with the second flow path to accumulate the bio material, and a second induction flow path connecting the second flow path and the second accumulation space, A second structure having a shape corresponding to the microchannel loading part is formed and a third protruding structure having a shape corresponding to the nano channel part connecting the first integrated space and the second integrated space is formed Step of making a control block for producing die with; The first protruding structure, the second protruding structure, and the third protruding structure are cured by supplying resin to the block making mold so that the first protruding structure, the second protruding structure and the third protruding structure are submerged, and then separated from the block manufacturing mold to form the micro / step; Preparing a substrate; And a surface of the micro / nano fluidic channel block facing the substrate with the first microchannel loading part, the second microchannel loading part, and the nano channel part facing the substrate, And adhering and fixing the biochemical reactor.

The biochemical reactor and the manufacturing method thereof according to the present invention have the following effects.

First, by connecting the first microchannel loading unit and the second microchannel loading unit to the nanochannel unit, diffusion due to the concentration difference occurs between the first microchannel loading unit and the second microchannel loading unit, Thereby allowing more accurate analysis of gene expression of microorganisms.

Secondly, since the diffusion rate from the second microchannel loading unit to the first microchannel loading unit can be adjusted linearly by controlling the number of formed nanochannel units, a more purified analysis becomes possible.

Thirdly, molds for making molds and molds for building blocks manufactured during the process of manufacturing biochemical reactors can be semi-permanently reused, which is advantageous in that a large number of biochemical reactors can be manufactured.

Fourthly, there is no deformation of initial designed dimension during manufacture of molds and blocks for manufacturing molds, so that a more accurate biochemical reactor can be manufactured.

1 and 2 show a structure of a biochemical reactor according to an embodiment of the present invention.
3 to 6 show a process of manufacturing the biochemical reactor according to FIG.
FIG. 7 is a photograph of the depths of the first micro-grooves, the first protruding structures and the nano-channels in the process of manufacturing the biochemical reactor according to FIGS.
FIG. 8 is a graph illustrating the depth and width of the first micro-grooves according to the number of the first micro-grooves formed in the mold for manufacturing a mold manufactured according to FIG.
FIG. 9 is a graph illustrating the depth and width of the first micro-grooves according to the length of the first micro-grooves in the mold for manufacturing a mold according to FIG.
FIG. 10 is a photograph and a graph of an experiment on genetic expression of microorganisms according to the difference in the number of nano-channels formed using the biochemical reactor according to FIG.
FIG. 11 is a photograph and a graph of an experiment on the genetic expression of microorganisms according to the length of the nanochannel using the biochemical reactor according to FIG.
FIG. 12 and FIG. 13 are photographs and graphs showing the genetic expression of microorganisms according to different embodiments according to the difference in the number of nano channel sections formed using the biochemical reactor according to FIG.
FIG. 14 and FIG. 15 are photographs and graphs illustrating the genetic expression of microorganisms according to the difference in the number of nano channel sections formed using the biochemical reactor according to another embodiment of the present invention.

1 to 15 show a biochemical reactor according to the present invention and a method for producing the same.

First, referring to FIGS. 1 and 2, a biochemical reactor according to an embodiment of the present invention includes a substrate (not shown), a micro / nano fluid (not shown) provided on the substrate And a channel block 100. The substrate (not shown) is bonded to the micro / nano fluid channel block 100 by bonding, for example, to a glass substrate. In this embodiment, a glass substrate is used, but the present invention is not limited thereto, and various substrates can be used.

The micro / nanofluid channel block 100 includes a first microchannel loading unit 110, a second microchannel loading unit 130, and a nanochannel unit 150. The first microchannel loading unit 110 includes a first flow path 111, a first guide path 115, and a first accumulation space 113. The first accumulation space 113 is connected to the first flow path 111 and the first accumulation space 113 is connected to the first flow path 111, 111) of the fluid flowing through the microbes (1). In the present embodiment, the first accumulation space 113 is formed in a semicircular shape, but it is not limited thereto and may be formed in various shapes.

The first induction passage 115 connects the first passage 111 and the first accumulation space 113 such that the first passage 111 communicates with the first accumulation space 113, The microbes 1 may be moved to the first accumulation space 113 in the fluid flowing through the first flow path 111.

In more detail, the first induction passage 115 includes a first induction passage 115a and a first auxiliary passage 115b. The first inflow passage 115a connects the first flow passage 111 and the first accumulation space 113 and the microorganisms 1 flowing through the first flow passage 111 are connected to the first accumulation space 113. [ (113).

The first auxiliary flow paths 115b face each other with respect to the first inflow path 115a on the first inflow path 115a and are spaced apart from each other along the longitudinal direction of the first inflow path 115a Are formed. Each of the first auxiliary flow paths 115b is inclined toward the first accumulation space 113 to store the microorganisms 1 separated from the first accumulation space 113,

The first induction passage 115 including the first induction passage 115a and the first auxiliary passage 115b is formed in a ratchet shape and the first accumulation space 113 can be directed only toward the first accumulation space (113).

When a plurality of the first accumulation spaces 113 are formed, at least one first accumulation space 113 is formed. The plurality of first accumulation spaces 113 are spaced apart from each other along the flow direction of the fluid flowing through the first flow path 111, . In the present embodiment, as shown in FIG. 1, three first integrated spaces 113 are formed as an example.

The second microchannel loading part 130 includes a second flow path 131, a second guide path 135, and a second accumulation space 133. The second flow path 131 is formed so that a fluid containing the bio material 3 for gene expression of the microorganisms 1 can be supplied and flowed, (3) of the fluid flowing along the second flow path (131) is connected to the first flow path (131). In the present embodiment, the second accumulation space 133 is formed in a semicircular shape like the first accumulation space 113, but may be formed in various shapes.

In the above description, the fluid containing the biomaterial is supplied to the second microchannel loading unit 130. However, the present invention is not limited thereto, and a fluid containing a microorganism capable of generating a biomaterial may be supplied. This will be described later in more detail.

The second induction passage 135 connects the second passage 131 and the second accumulation space 133 such that the second passage 131 and the second accumulation space 133 communicate with each other, (3) so that the bio-material (3) can be applied to the second accumulation space (133) in the fluid flowing through the second flow path (131).

The second induction passage 135 connects the second passage 131 and the second accumulation space 133 so that the second passage 131 and the second accumulation space 133 are connected to each other, Like the first induction passage 115, includes a second induction passage 135a and a second auxiliary passage 135b. The second induction passage 135 is connected to the second accumulation space 133 in such a manner that the biomaterial 3 can be transferred to the second accumulation space 133 from among the fluid flowing through the second flow path 131, .

The structure of the second induction passage 135 is the same as that of the first induction passage 115 as shown in FIG. The second inlet passage 135a connects the second flow passage 131 and the second accumulation space 133 and the bio-materials 3 of the fluid flowing through the second flow passage 131 are connected to the second flow- 2 accumulation space 133, as shown in FIG.

The second auxiliary flow paths 135b face each other with respect to the second inflow path 135a on the second inflow path 135a and are spaced apart from each other along the longitudinal direction of the second inflow path 135a Are formed. Each of the second auxiliary channels 135b is inclined toward the second accumulation space 133 to store the bio materials 3 separated from the second accumulation space 133. [

The second induction passage 135 including the second inflow channel 135a and the second bypass channel 135b is formed in a ratchet shape and the bio material 3 is formed by the ratchet shape, May be transferred to the second accumulation space 133 and applied to the second accumulation space 133.

At least one second accumulation space 133 is formed in the same manner as the first accumulation space 113. When a plurality of the second accumulation spaces 133 are formed, And are arranged in parallel to each other along the flow direction of the fluid. The positions of the second accumulation space 133 are symmetrical with the positions of the first accumulation space 113 and are formed in the same number as the first accumulation space 113.

The nano channel part 150 connects the first accumulation space 113 and the second accumulation space 133 such that the first accumulation space 113 and the second accumulation space 133 communicate with each other. The bio-materials 3 accumulated in the second accumulation space 133 by the nano-channel unit 150 are supplied to the microorganisms 1 accumulated in the first accumulation space 113. That is, the bio materials 3 are transferred to the first accumulation space 113 along the nano channel part 150 and supplied to the microorganisms 1.

The depth of the nano channel part 150 is set within a range of 1 nm to 1000 nm, the width is set within a range of 1 to 10 mu m, and the length is set within a range of 1 to 1000 mu m. The nano channel part 150 according to this embodiment is formed to have a depth of 330 nm, a width of 3.4 m, and a length of 200 m, for example. The formation of the nano channel part 150 may be performed by controlling the amount of the convective current between the first accumulation space 113 and the second accumulation space 133 so that sufficient diffusion can be achieved .

When a plurality of the first integrated space 113 and the second integrated space 133 are provided as described above, the nano channel unit 150 may be formed in various embodiments. First, when a plurality of the first integrated space 113 and the second integrated space 133 are provided, the lengths of the nano channel units 150 are the same, but each of the first integrated space 113 and the second integrated space 133 The number of the second integrated spaces 133 may be different from each other.

More specifically, in the present embodiment, as described above, the three first integrated spaces 113 and the three second integrated spaces 133 are provided, and one of the first integrated spaces 113 and One nano channel part 150 is formed between the second integrated spaces 133 which are symmetric. On the other hand, three nano channel parts 150 are formed between the first integrated space 113 and the second integrated space 133 which are symmetrical with each other. On the other hand, five nano channel parts 150 are formed between the second integrated space 133 which is symmetrical with another one of the first integrated spaces 113.

On the other hand, when a plurality of the first accumulation space 113 and the second accumulation space 133 are provided, the nano- The number of channel units 150 is the same, but the lengths thereof may be different.

More specifically, when three of the first integrated spaces 133 and three of the second integrated spaces 133 are provided, the first integrated space 113 and the second integrated space 133, which are illustratively symmetric with respect to the first integrated space 113, The length of the nano channel part 150 connecting the second integrated space 133 is 100 m and the length of the nano channel part 150 connecting the second integrated space 133, which is symmetric with the other first integrated space 113, The length of the channel section 150 is 200 mu m and the length of the nano channel section 150 connecting the second accumulation space 133 symmetrical to the other one of the first accumulation spaces 113 is 400 mu m to be.

As described above, the nano channel part 150 may be formed in various embodiments, and the formation of the nano channel part 150 may be variously modified according to the experiment of gene expression of the microorganism.

3 to 5 show a method of manufacturing the micro / nano fluidic channel block 100 as described above. In order to manufacture the micro / nano fluid channel block 100, a block mold 40 for manufacturing the micro / nano fluid channel block 100 is first fabricated. The block making mold 40 includes a first protruding structure 41 corresponding to the first microchannel loading unit 110, a second protruding structure 43 corresponding to the second microchannel loading unit 130, And a third protruding structure 45 corresponding to the nano channel part 150 is formed. This will be described later in more detail.

When the mold 40 for forming a block is manufactured, the micro / nano fluidic channel block 100 is formed by applying resin to the block making mold 40 and curing the mold 40 and then separating the block making mold 40 from the block making mold 40. The surface of the micro / nano fluidic channel block 100 formed with the first microchannel loading unit 110, the second microchannel loading unit 130, and the nanochannel unit 150, To adhere the micro / nano fluidic channel block 100 to the substrate.

The block molding die 40 must be manufactured before the micro / nano fluid channel block 100 is manufactured and the mold 30 for manufacturing the block molding die 40 is manufactured. Should be done first. That is, the mold for manufacturing a mold 30 is firstly manufactured first, and then the mold 40 for molding is manufactured using the mold 30 for manufacturing a mold. Then, the micro / nano fluid channel block 100 is fabricated using the block making mold 40.

A first micro hole 31 corresponding to the first projecting structure 41 and a second micro hole 33 corresponding to the second projecting structure 43 are formed on the upper surface of the die 30 for mold- 3 protruding structures 45 are formed in the first micro-grooves 35. As shown in FIG. The method of manufacturing the mold 30 for molding the mold will be described later in more detail.

When the mold 30 for manufacturing a mold is manufactured, a resin is filled in the first fine holes 31, the second fine holes 33, and the first fine grooves 35 of the mold 30, And then separated from the mold 30 for forming a mold to form the mold 40 for making a block.

A method of manufacturing the mold 30 for molding the above-described mold will be described in more detail with reference to FIG. In order to manufacture the mold 30, the photosensitive material is coated on the substrate 10 by a spin process and then baked in an oven to cure the photosensitive material to form a photosensitive material layer 30a. The photosensitive material is exemplarily SU-8 polymer. And a first photomask 20 applied with the first pattern 21 corresponding to the first microhole 31 and the second microhole 33 formed on the photosensitive material layer 30a , The photosensitive material layer 30a is primarily exposed by irradiating the light energy. Although the first pattern 21 is simply represented in the drawing, the first pattern 21 may be formed by the first microchannel loading unit 110, the second microchannel loading unit 13, And are formed in the same shape.

The SU-8 polymer applied to the photosensitive material layer 30a is a negative photosensitive material. The portion covered with the first pattern 21 is not cured, and the portion irradiated with the light energy is cured. The light energy for exposing the photosensitive material layer 30a is exemplified by ultraviolet (UV) light. At this time, the light energy can use the entire ultraviolet (UV) wavelength band, or a combination of the set long wavelength and the set short wavelength.

On the other hand, a cross linking gradient is formed along the depth direction of the portion exposed by the light energy. The SU-8 polymer forming the photosensitive material layer 30a includes a monomer unit. When the light energy is irradiated, the monomer is concentrated in the direction in which the light energy is irradiated. Therefore, in the region where the monomer is concentrated, the space between the monomers is reduced to have an elastic property, and in a region where the monomer is not concentrated, a space is increased between the monomers and the polymer has viscoelastic properties. Thus, a cross linking gradient is formed along the depth direction in the photosensitive material layer 30a.

When the photosensitive material layer 30a that has been primarily exposed is immersed in a developing solution and developed, the portion where the light energy is not irradiated by the first pattern 21 is removed by the developing solution, and the first fine holes 31, And the second fine holes 33 are formed. A second photomask 20a is provided on the photosensitive material layer 30a on which the first microhole 31 and the second microhole 33 are formed and the second exposure is performed by irradiating the light energy. At this time, a second pattern 41 corresponding to the setting area including the first fine groove 35 is formed in the second photomask 20a. Accordingly, the photosensitive material layer 30a is cured by irradiating the light energy only to a portion except the region corresponding to the second pattern 41. [

When the photosensitive material layer 30a having been secondarily exposed is immersed in a developing solution and developed, the first fine holes 31 or the second fine holes 31 are formed in the portions to which the light energy is not irradiated by the second pattern 41 The cracks are generated from any one of the fine holes 31 and 33 and the crack 35a proceeds to the fine one of the first fine hole 31 and the second fine hole 33, 1 fine grooves 35 formed in the surface of the substrate.

Since the crack 35a generated by the secondary exposure of the photosensitive material layer 30a proceeds only in a region corresponding to the second pattern 41, the crack 35a is prevented from proceeding in an undesired direction can do. Also, even if a new crack 35a is generated in the crack 35a, it is possible to prevent the newly generated crack 35a from proceeding too much.

As shown in FIG. 4, any one of the first micro holes 31 or the second micro holes 33, in which the crack 35a is generated, A notch 33a extending along the height direction of the photosensitive material layer 30a in the direction toward the other one of the fine holes is formed. The notch 33a is formed in a triangular or quadrangular shape in cross section, and is formed in a triangular shape as shown in FIG. 4 in the present embodiment.

On the other hand, the crack 35a is formed by an angle formed by one surface 33a-1 of the notch 33a and the other surface 33a-2 of the notch 33a (hereinafter referred to as an angle? Of the notch) The width-to-height contrast, and the primary exposure time of the photosensitive material layer 30a. The method of manufacturing the mold for manufacturing a mold as described above may be manufactured by applying a method of manufacturing a microchannel according to Japanese Patent Application No. 10-1390700 filed by the applicant of the present invention.

In the present embodiment, the photosensitive material layer 30a is exposed twice and developed twice to produce the mold. On the other hand, according to the method of the above-mentioned Japanese Patent No. 10-1390700, It is possible to manufacture the mold for manufacturing the mold. In addition, the above-mentioned Japanese Patent No. 10-1390700 discloses a technique for controlling the formation of the crack by using the angle of the notch, the contrast of the width and height of the notch, and the primary exposure time of the photosensitive material layer 30a The formation of the first fine grooves 35 can be controlled by applying this feature.

As shown in FIG. 5, the resin is supplied to the die 30 for manufacturing a mold manufactured by the above-described method, and then the resin is cured. Then, the mold 30 is separated from the mold 30 for manufacturing a block, You can. The resin supplied to the mold 30 for forming a mold is exemplarily polyurethane (PUA).

When the mold 30 is supplied with the PUA, the PET film 50 is covered on the PUA. The PUA supplied to the mold 30 for molding the mold is filled in the first microhole 31, the second microhole 33 and the first microhole 35, A first protruding structure 41 corresponding to the first fine hole 31, the second fine hole 33 and the first fine groove 35, the second protruding structure 43, The protruding structure 45 is formed.

6, the polymethylsiloxane (PDMA) is supplied to the mold 40 for block manufacture and cured. When the mold 40 is separated from the mold 40, the micro / The manufacture of the block 100 is completed.

The mold 30 and the block making mold 40 manufactured by the above-described method can be reused many times once they are fabricated, so that a large number of micro / nano fluidic channel blocks 100 can be produced for a short time , Thereby enabling the production of a large amount of biochemical reactors.

Particularly, the depth of the nano channel part 150 formed in the micro / nano fluid channel block 100 manufactured according to the method described above corresponds to the nano channel part 150 as shown in FIG. 7 The micro / nano fluidic channels 45 and the first micro / nano fluid channels 46 are not significantly different in size as compared with the first micro / trenches 33 corresponding to the third protruding structures 45 and the third protruding structures 45, It can be confirmed that the manufacturing method of the block 100 has high accuracy.

Referring to FIG. 7, (a) shows the depth of the first fine groove 35 formed in the mold 30 for molding the die, and the depth of the first fine groove 35 is 340 nm. (b) shows the height of the third projecting structure 45 formed on the block making mold 40 manufactured by the mold 30, and the height of the third projecting structure 45 is 335 nm . (c) is a measurement of the depth of the nano channel part 150 of the micro / nano fluidic channel block 100 manufactured by the block making mold. The depth of the nano channel part 150 is 332 nm. Thus, it can be seen that the depth of the nano channel part 150 is reduced by about 2% as compared with the depth of the first fine groove 35, which is within an error range. In this way, it can be confirmed that the shape is highly accurate even in a large number of manufacturing processes.

As described above, when the first accumulation space 113 and the second accumulation space 133 are provided in plurality, the nano-channel unit 150 may include the first accumulation space 113 and the second accumulation space 133, The number of the nano channel parts 150 may be different even if the lengths of the nano channel parts 150 connecting the second integrated spaces 133 are the same. Even when a plurality of cracks 35a are simultaneously formed in the process of forming the cracks 35a, the depth and width of the cracks 35a are substantially the same.

8 shows the results of measuring the depth and width of the cracks according to the number of cracks connecting the first fine holes and the second fine holes. As shown in the graph of FIG. 8 (b) (A3), the depth of the cracks was almost the same, and the width of the cracks was also within an error range (A1). In the case where one crack was formed (A2) .

When a plurality of the first accumulation space 113 and the second accumulation space 133 are provided, the distance between the first accumulation space 113 and the second accumulation space 133 The distance between the first integrated space 113 and the second integrated space 133 may be different from each other. For this purpose, the lengths of the cracks are differently formed in a process of forming cracks for forming the nano channel part 150.

9 is a graph showing the results of measuring the depth and width of the cracks when the lengths of the cracks connecting the first fine holes and the second fine holes are different from each other. 9 (a), the length of the crack gradually becomes longer from left to right (L1 <L2 <L3). Thus, when the lengths of the cracks are all different, the depth and width of the cracks It can be seen that the depth and the width are formed almost similar regardless of the length of the crack.

According to the measurement result, the diffusion of the bio-material by the nano channel unit 150 can be controlled by using the number of the nano channel unit 150 and the length of the nano channel unit 150 I can do it.

FIGS. 10 and 11 are graphs showing the results of a biochemical reactor produced by the above- FIG. 10 is a graph illustrating the diffusion rate according to the number of the nano channel units 150, and FIG. 11 is a graph illustrating the diffusion rate according to the length of the nano channel unit 150. FIG. will be.

For the experiment, 100 μM of FITC (fluorescein isothiocyanate) was added to PBS (Phosphate buffer saline) solution, and the resultant was supplied to the second microchannel loading unit 130, Only the pure PBS solution was supplied to the portion 110. In the PBS solution supplied to the second microchannel 130, the FITC is introduced into the second accumulation space 133 and accumulated in the second accumulation space 133.

The first microchannel loading unit 110 and the second microchannel loading unit 130 are diffused due to the difference in concentration due to the FITC. That is, diffusion of the FITC included in the PBS solution supplied to the second microchannel loading unit 130 to the first microchannel loading unit 110 through the nanochannel unit 150 occurs.

10 (b) is a graph showing the fluorescence intensities of the first microchannel loading unit 110 measured in the first accumulation space 113 as the FITC moves to the first microchannel loading unit 110 As a result, it can be seen that the brightness of the AA 'portion of the first accumulation space 113 becomes brighter with time. Particularly, it can be confirmed that the fluorescence intensity of 40 minutes after the start of the experiment sharply increases and that the fluorescence intensity does not increase from 40 minutes after the experiment begins. After 40 minutes, the first microchannel loading unit 110 And the concentration difference of the second microchannel loading part 130 decrease remarkably after 40 minutes. In addition, it can be seen that the brightness of the first accumulation space 113, A3 in which five of the three first accumulation spaces 113 are formed is the brightest. That is, by controlling the number of the nano channel units 150, the diffusion rate can be controlled. In particular, the number of the nano channel units 150 can be adjusted linearly.

11A and 11B illustrate an experiment to determine the difference in diffusion speed according to the length of the nano channel part 150. Referring to FIG. 11A, the first integrated space 113 And the nano channel part 150 connecting the second accumulation space 133 may be formed in a longer length than the nano channel part 150 from the left to the right.

11 (b) is a graph showing the diffusion rate measured in the biochemical reactor as shown in FIG. 11 (a). As shown in FIG. 11 (b) It can be seen that the shorter the length, the more the diffusion becomes. However, when the number of the nano channel parts 150 is the same and the length of the nano channel part 150 is different, the degree of diffusion of the nano channel part 150 into each of the first accumulation spaces 113 It is effective to control the gene expression rate of the microorganism using the above-described biochemical reactor by using the number of the nanostructured channels 150.

12 to 15 are diagrams illustrating a case where the lengths of the nano channel parts 150 connecting the first integrated space 113 and the second integrated space 133 are the same, (1) using the above-mentioned biochemical reactors which are different from each other.

12 shows that receiver cells (RC) are integrated in the first integrated space 113 of the first microchannel loading unit 110 and the receiver cells RC are integrated in the second integrated channel 113 of the second microchannel loading unit 110, And acyl homoserine lactone (hereinafter referred to as AHL) is applied to the reaction chamber 133.

The first accumulation space 113 and the second accumulation space 133 are formed such that the AHL of the second accumulation space 133 is separated from the first accumulation space 133 through the nano- Space 113 and supplied to the recipient cell 1. The recipient cell 1 receives the AHL (3) and produces a green fluorescent protein (GFP). The gene expression rate of the recipient cell (1) can be confirmed by the green fluorescent protein produced by the recipient cell (1).

Referring to FIG. 12, when the photograph of the first accumulation spaces 113 taken in time is taken, it can be seen that as the time increases, the gene expression of the recipient cell RC increases and the green fluorescent protein increases. In particular, it can be seen that the more the number of the nano channel part 150 is formed, the more the gene expression of the recipient cell RC is performed and the green fluorescence protein is also generated more.

FIG. 13 is a graph showing the experimental results of FIG. 12, showing that the production of the fluorescent green protein is increased due to gene expression of the microorganisms 1 accumulated in the first integrated space 113 with time It can be confirmed that the fluorescent green protein is produced more as the number of the nano channel part 150 is increased.

14, the recipient cells 1 are integrated in the first integrated space 113 of the first microchannel loading unit 110 and the recipient cells 1 are integrated in the second microchannel loading unit 130. In addition, In the second accumulation space 133, the transmitter cells 5 generating the AHL are integrated. The AHL generated in the transmitter cell 5 is supplied to the receiver cells 1 integrated in the first accumulation space 113 through the nanochannel unit 150. 14, the recipient cells 1 receive the AHL generated in the sender cell 5 and generate the green fluorescent protein.

As shown in FIG. 15, which graphically shows the photographic image of FIG. 14 and the experimental result of FIG. 14, as the number of formed nanotubes 150 is larger, the recipient cell 1 has more green fluorescence It can be confirmed that the protein is produced. 14, the supply of the biomaterial for the genetic expression of the microorganism 1 integrated in the first accumulation space 113 can be performed by genetic induction between the microorganisms . 14 does not directly supply the biomolecule to the recipient cell 1 but receives the biomaterial through the sender cell 3 so that the recipient cell 1 and the sender cell 3 are not biomaterial- It can be confirmed that the biomaterial is supplied by genetic induction between the cells. Therefore, in the biochemical reactor, the second microchannel loading unit 130 may be supplied with a fluid containing a microorganism that can be genetically induced, rather than the biomaterial.

As described above, the biochemical reactor according to the present embodiment is an optimal device for testing the genetic expression of microorganisms through various experiments, and it is possible to control the gene expression rate of the microorganisms by controlling the number of the formation of the nano channel part 150 And thus it is possible to perform more accurate analysis.

While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. Accordingly, the true scope of the present invention should be determined by the technical idea of the appended claims.

10: substrate 30a: photosensitive material layer
20: first photomask 20a: second photomask
30: mold for making mold 40: mold for making block
100: micro / nanofluid channel block
110: first microchannel loading part 111: first microchannel loading part
113: first accumulation space 115: first induction flow path
130: second microchannel loading part 131: second microchannel loading part
133: second accumulation space 135: second induction flow path

Claims (18)

delete delete delete delete delete delete delete A first flow path through which a fluid containing microorganisms is supplied and flows, a first integrated space in which the microorganisms are integrated by being communicated with the first flow path, and a second integrated path through which the first flow path and the first integrated space are connected A second flow path in which a first protruding structure having a shape corresponding to a first microchannel loading part formed with an induction flow path is formed and spaced apart from the first protrusion structure and a fluid containing a bio material is supplied and flows, A second protrusion having a shape corresponding to a second microchannel loading section in which a second microchannel loading section in which a second microchannel loading section in which the second microchannel loading section and the second microchannel are connected is formed, And a third protruding structure having a shape corresponding to the nano channel part connecting the first integrated space and the second integrated space is formed, Step;
Forming a micro / nanofluid channel block by supplying resin to the block making mold so that the first protruding structure, the second protruding structure, and the third protruding structure are submerged and curing, and then separating the block from the block making mold; ;
Preparing a substrate; And
In the micro / nano fluidic channel block, the first microchannel loading unit, the second microchannel loading unit, and the surface having the nano channel part are disposed facing the substrate to adhere the micro / nano fluid channel block and the substrate And fixing,
Before the step of producing the block making mold,
A first fine hole having a shape corresponding to the first projecting structure on the upper surface, a second fine hole having a shape corresponding to the second projecting structure, and a first fine groove having a shape corresponding to the third projecting structure are formed A step of fabricating a mold for manufacturing a mold,
The step of fabricating the mold for manufacturing a mold,
Applying and curing a photosensitive material on a substrate to form a photosensitive material layer;
A first photomask having a first pattern on the photosensitive material layer and having a first pattern corresponding to the first microhole and the second microhole formed on the photosensitive material layer;
Forming the first microhole and the second microhole by removing a region corresponding to the first pattern while developing the photosensitive material layer that has been primarily exposed;
And a second photomask having a second pattern corresponding to a setting region including the first fine groove on the photosensitive material layer on which the first fine holes and the second fine holes are formed, A second exposure step; And
The method comprising the steps of: developing a secondarily exposed photosensitive material layer to generate cracks in one of the first fine holes or the second fine holes while developing the other fine holes of the first fine holes or the second fine holes And advancing the crack toward the hole to form the first fine groove. delete The method of claim 8,
The step of fabricating the block-
The resin is supplied to be filled in the first fine hole, the second fine hole, and the first fine groove on the mold for manufacturing a mold, and then the resin is cured. Then, the resin is molded and separated from the mold for manufacturing a mold, Lt; / RTI &gt; delete The method of claim 8,
Wherein a notch is formed in any one of the first microhole and the second microhole so that the crack can be generated. The method of claim 12,
Wherein the number of cracks is controlled by adjusting an angle of the notch formed between the one surface of the notch and the other surface of the notch. The method of claim 12,
The notch has a triangular shape having a width and a height,
Wherein the number of occurrences of the cracks is adjusted by using the phase contrast between the width and the height. The method of claim 8,
In the primary exposure step,
Wherein a region of the photosensitive material layer not corresponding to the first pattern is changed in elastic property to a viscoelastic property along a depth direction. The method of claim 8,
Wherein the photosensitive material is a negative photosensitive material which is cured by light, and comprises a SU-8 polymer. The method of claim 10,
Wherein the resin supplied to the mold for molding in the step of manufacturing the mold for block making comprises polyurethane. The method of claim 8,
Wherein the resin supplied to the mold for block making comprises polydimethylsiloxane (PDMS).
KR1020150147028A 2015-10-22 2015-10-22 Manufacturing method of biochemical reactors KR101716302B1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
KR1020150147028A KR101716302B1 (en) 2015-10-22 2015-10-22 Manufacturing method of biochemical reactors
PCT/KR2016/005639 WO2017069364A1 (en) 2015-10-22 2016-05-27 Biochemical reactor and manufacturing method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1020150147028A KR101716302B1 (en) 2015-10-22 2015-10-22 Manufacturing method of biochemical reactors

Publications (1)

Publication Number Publication Date
KR101716302B1 true KR101716302B1 (en) 2017-03-27

Family

ID=58496832

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1020150147028A KR101716302B1 (en) 2015-10-22 2015-10-22 Manufacturing method of biochemical reactors

Country Status (2)

Country Link
KR (1) KR101716302B1 (en)
WO (1) WO2017069364A1 (en)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20200090506A (en) * 2019-01-21 2020-07-29 울산과학기술원 Self-powered diffusiophoresis apparatus and method of performing self-powered diffusiophoresis using the same
KR20200090507A (en) * 2019-01-21 2020-07-29 울산과학기술원 Method of performing self-powered diffusiophoresis using the same
KR102218278B1 (en) * 2019-11-08 2021-02-19 울산과학기술원 Apparatus for controlling the transport of materials in nanochannels by controlling humidity
KR20210048276A (en) * 2019-10-23 2021-05-03 울산과학기술원 Apparatus for separating nanoparticles and methods for separating nanoparticles using the apparatus
WO2021101135A1 (en) * 2019-11-19 2021-05-27 울산과학기술원 Micro-object extraction method using diffusiophoresis, and micro-object identification method using same
KR20230043460A (en) * 2021-09-24 2023-03-31 울산과학기술원 Microfluidic film and method for fabricating the microfluidic film
KR20230043462A (en) * 2021-09-24 2023-03-31 울산과학기술원 Microfluidic module and method for fabricating the microfluidic module

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101206619B1 (en) 2010-04-06 2012-11-29 국립대학법인 울산과학기술대학교 산학협력단 Microfluidic screening device
KR101390700B1 (en) * 2013-07-15 2014-04-30 국립대학법인 울산과학기술대학교 산학협력단 Method for making fine channel

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101125060B1 (en) * 2009-07-22 2012-03-21 한국과학기술원 Microfluidic device of capturing particles and method of capturing particles using it
KR101140486B1 (en) * 2010-03-30 2012-04-30 국립대학법인 울산과학기술대학교 산학협력단 Microfluidic concentrator
KR101330473B1 (en) * 2011-07-25 2013-11-15 국립대학법인 울산과학기술대학교 산학협력단 Microfluidic concentrator for communication assays of microbes

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101206619B1 (en) 2010-04-06 2012-11-29 국립대학법인 울산과학기술대학교 산학협력단 Microfluidic screening device
KR101390700B1 (en) * 2013-07-15 2014-04-30 국립대학법인 울산과학기술대학교 산학협력단 Method for making fine channel

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Microfabricated ratchet structure integrated concentrator arrays for synthetic bacterial cell-to-cell communication assays(Lab chip, 2012)* *
단순공정으로 제작된 마이크로나노 하이브리드 채널의 불균형 동전기성을 이용한 미세혼합기 연구(대한기계학회논문집, 2011)* *

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20200090506A (en) * 2019-01-21 2020-07-29 울산과학기술원 Self-powered diffusiophoresis apparatus and method of performing self-powered diffusiophoresis using the same
KR20200090507A (en) * 2019-01-21 2020-07-29 울산과학기술원 Method of performing self-powered diffusiophoresis using the same
KR102168202B1 (en) 2019-01-21 2020-10-20 울산과학기술원 Method of performing self-powered diffusiophoresis using the same
KR102168201B1 (en) 2019-01-21 2020-10-20 울산과학기술원 Self-powered diffusiophoresis apparatus and method of performing self-powered diffusiophoresis using the same
KR102299472B1 (en) * 2019-10-23 2021-09-07 울산과학기술원 Apparatus for separating nanoparticles and methods for separating nanoparticles using the apparatus
KR20210048276A (en) * 2019-10-23 2021-05-03 울산과학기술원 Apparatus for separating nanoparticles and methods for separating nanoparticles using the apparatus
WO2021091203A1 (en) * 2019-11-08 2021-05-14 울산과학기술원 Device for controlling material delivery in nanochannels through humidity control
KR102218278B1 (en) * 2019-11-08 2021-02-19 울산과학기술원 Apparatus for controlling the transport of materials in nanochannels by controlling humidity
WO2021101135A1 (en) * 2019-11-19 2021-05-27 울산과학기술원 Micro-object extraction method using diffusiophoresis, and micro-object identification method using same
KR20210060945A (en) * 2019-11-19 2021-05-27 울산과학기술원 Method for extracting fine object using diffusiophoresis and identification method of the fine object using the method
KR102299473B1 (en) * 2019-11-19 2021-09-07 울산과학기술원 Method for extracting fine object using diffusiophoresis and identification method of the fine object using the method
KR20230043460A (en) * 2021-09-24 2023-03-31 울산과학기술원 Microfluidic film and method for fabricating the microfluidic film
KR20230043462A (en) * 2021-09-24 2023-03-31 울산과학기술원 Microfluidic module and method for fabricating the microfluidic module
KR102558147B1 (en) * 2021-09-24 2023-07-20 울산과학기술원 Microfluidic film and method for fabricating the microfluidic film
KR102600749B1 (en) * 2021-09-24 2023-11-09 울산과학기술원 Microfluidic module and method for fabricating the microfluidic module

Also Published As

Publication number Publication date
WO2017069364A1 (en) 2017-04-27

Similar Documents

Publication Publication Date Title
KR101716302B1 (en) Manufacturing method of biochemical reactors
Lee et al. Three-dimensional fabrication of heterogeneous microstructures using soft membrane deformation and optofluidic maskless lithography
Kim et al. Soft lithography for microfluidics: a review
Kim et al. Cell research with physically modified microfluidic channels: a review
Amato et al. Integrated three-dimensional filter separates nanoscale from microscale elements in a microfluidic chip
Khoury et al. Ultra rapid prototyping of microfluidic systems using liquid phase photopolymerization
JP4931330B2 (en) Microfluidic structure, in particular a method for producing a biochip and the structure obtained by said method
US20100099048A1 (en) Stop Flow Interference Lithography System
US9404914B2 (en) Microfluidic system for controlling a concentration profile of molecules capable of stimulating a target
Morel et al. Concentration landscape generators for shear free dynamic chemical stimulation
Liu et al. A facile “liquid-molding” method to fabricate PDMS microdevices with 3-dimensional channel topography
CN112892627B (en) Photocuring micro-fluidic chip based on elastic support body and preparation method and application thereof
RU2748273C9 (en) Nano-imprinted substrate
US20100207301A1 (en) Method of forming fine channel using electrostatic attraction and method of forming fine structure using the same
Gao et al. Direct optical micropatterning of poly (dimethylsiloxane) for microfluidic devices
KR101390665B1 (en) Concentration gradient generating microfluidic device and method of manufacturing the same
US20140191438A1 (en) Microfluidic Devices and Methods of Fabrication
KR101390700B1 (en) Method for making fine channel
KR100969551B1 (en) Method of forming fine channel using electrostatic attraction and method of forming fine structure using the same
KR101878936B1 (en) Method for making crack of micro channel
CN109821583B (en) Processing method of special-shaped cross section micro-channel chip based on FDM three-dimensional printing
KR20040009187A (en) Method of manufacturing microchip
Zhang et al. An optically fabricated gradient nanochannel array to access the translocation dynamics of T4-phage DNA through nanoconfinement
Li 3D Printed Microfluidic Devices for Controlled Biomaterial Fabrication
De Beer Advances in additive manufacturing and microfabrication

Legal Events

Date Code Title Description
E701 Decision to grant or registration of patent right
GRNT Written decision to grant