KR102009563B1 - Low temperature flow lithography method and low temperature flow lithography system - Google Patents

Abstract

The present invention relates to a low temperature fluid lithography method and a low temperature fluid lithography system, wherein the low temperature fluid lithography method comprises (a) flowing a photocurable fluid 1 into a microchannel 10 (S100), and (b) irradiating the microchannel 10 with light 3 to synthesize the fine particles 5 (S200), wherein steps (a) and (b) have a temperature relatively higher than room temperature. It is run at low temperature synthesis temperatures.

Description

LOW TEMPERATURE FLOW LITHOGRAPHY METHOD AND LOW TEMPERATURE FLOW LITHOGRAPHY SYSTEM

The present invention relates to a low temperature fluid lithography method and a low temperature fluid lithography system, and more particularly, to a technique for synthesizing microparticles by performing fluid lithography at a low temperature below room temperature.

Microparticles are used in various fields such as tissue engineering, drug delivery, and bioassay. Techniques for synthesizing such microparticles include emulsion polymerization, microfluidic techniques, printing techniques, and fluid lithography. Fluid lithography is a technique for synthesizing particles through a radical reaction by irradiating ultraviolet light to a fluid flowing inside a microchannel, as disclosed in a patent document in the following prior art document. Here, when the ultraviolet rays to be irradiated pass through the mask punctured in a specific pattern, particles having a cross section corresponding to the puncture pattern may be generated. At this time, the height of the particles coincides with the height of the channel. In addition, different fluids may flow side by side within the microchannel to generate particles having various types of polymer regions.

Particles synthesized by such fluid lithography are formed by polymerizing photocurable monomers that are precursors of the particles into a network structure. Here, the synthesized particles may be added to the functional material in the form of being copolymerized to the net, or mounted between the net.

However, the conventional fluid lithography synthesizes the particles in a very short time at room temperature, so that the degree of polymerization of the particles is very low. Since the degree of polymerization indicates the degree of formation of the particle network, a low degree of polymerization makes it impossible to obtain the desired particle size, firmness, elastic modulus, and geometry. In addition, when the degree of polymerization is low, it is difficult to bind or mount the functional material on the particles. In order to increase the degree of polymerization, the synthesis time or the amount of UV irradiation must be increased. In this case, the particle productivity decreases, and the defect rate of the particles sticking to the microchannels increases.

Therefore, there is an urgent need for a solution for solving the problems of the conventional fluid lithography.

KR 10-2008-0070684 A

The present invention is to solve the above-mentioned problems of the prior art, an aspect of the present invention is to provide a technique for synthesizing microparticles by performing fluid lithography at a low temperature, relatively lower than room temperature.

The low temperature fluid lithography method according to the present invention comprises the steps of (a) flowing a photocurable fluid into a microchannel; And (b) irradiating light to the microchannels to synthesize fine particles, wherein the steps (a) and (b) are performed at a synthesis temperature of low temperature, which is relatively lower than room temperature. do.

Further, in the low temperature fluid lithography method according to the present invention, the synthesis temperature is 0 to 15 ° C.

Further, in the low temperature fluid lithography method according to the present invention, the photocurable fluid includes a photocurable monomer and a photoinitiator, and further includes at least one or more of a copolymer and nanoparticles.

In addition, in the low temperature fluid lithography method according to the present invention, the fine particles have a polyhedral shape having a height of 1 to 200 μm in cross section and a width of 1 to 500 μm in cross section.

In addition, in the low temperature fluid lithography method according to the present invention, the copolymer includes any one or more selected from the group consisting of fluorescent substance, DNA probe, antibody, virus, protein, and metal compound.

In addition, in the low temperature fluid lithography method according to the present invention, the nanoparticles include any one or more selected from the group consisting of nanofluorescent particles, nanomagnetic particles, proteins, nanometal particles, nanochemical particles, and cells.

On the other hand, the low temperature fluid lithography system according to the present invention includes a microfluidic chip having a microchannel through which the photocurable fluid flows; A light source for irradiating light to the microchannels to synthesize fine particles; And a cooling unit maintaining the temperature in the microchannel at a low temperature synthesis temperature relatively lower than room temperature.

In addition, in the low temperature fluid lithography system according to the present invention, the cooling unit comprises: a heat conducting plate in contact with an outer surface of the microfluidic chip; It includes; and a cooling device for cooling the heat conducting plate.

Further, in a low temperature fluid lithography system according to the present invention, the cooling unit comprises: a cooling fluid; And a cooling fluid circulation channel formed inside the microfluidic chip so that the cooling fluid flows.

Further, in a low temperature fluid lithography system according to the present invention, the cooling unit comprises: a cooling fluid; And a cooling fluid circulation tube in contact with an outer surface of the microfluidic chip and formed such that the cooling fluid flows.

In addition, in the low temperature fluid lithography system according to the present invention, the cooling unit has a double wall structure in which an inner wall and an outer wall are spaced apart from each other, and a container part in which the microfluidic chip is disposed in a space surrounded by the inner wall; And a coolant disposed between the inner wall and the outer wall.

Further, in the low temperature fluid lithography system according to the present invention, the synthesis temperature is 0 to 15 ° C.

The features and advantages of the present invention will become more apparent from the following detailed description based on the accompanying drawings.

Prior to this, the terms or words used in this specification and claims are not to be interpreted in a conventional and dictionary sense, and the inventors may appropriately define the concept of terms in order to best explain their invention in the best way possible. It should be interpreted as meaning and concept corresponding to the technical idea of the present invention based on the principle that the present invention.

According to the present invention, the fluid lithography is carried out at a temperature lower than room temperature, thereby improving the degree of polymerization of the particles, the particles have a more complete geometrical shape, and the functional material is copolymerized or mounted at a high rate to ensure improved functional sensitivity. have.

In addition, when the biomolecule detection probe is copolymerized at a low temperature to the particles, the detection sensitivity is increased and the detection limit is lowered, thereby saving the starting materials and improving the economic efficiency, as well as improving the performance of the functional particles.

1 is a process diagram of a low temperature fluid lithography method according to the present invention.
2 is a perspective view schematically showing a low temperature fluid lithography system according to a first embodiment of the present invention.
3 is a perspective view schematically showing a low temperature fluid lithography system according to a second embodiment of the present invention.
4 is a perspective view schematically showing a low temperature fluid lithography system according to a third embodiment of the present invention.
5 is a perspective view schematically showing a low temperature fluid lithography system according to a fourth embodiment of the present invention.
6 is a photograph of microparticles synthesized by a fluid lithography method.
7 to 10 are images and graphs showing fluorescence intensity according to synthesis temperature of microparticles synthesized by a fluid lithography method.

The objects, specific advantages and novel features of the present invention will become more apparent from the following detailed description and the preferred embodiments associated with the accompanying drawings. In the present specification, in adding reference numerals to the components of each drawing, it should be noted that the same components as possible, even if displayed on different drawings have the same number as possible. In the following description, detailed descriptions of related well-known techniques that may unnecessarily obscure the subject matter of the present invention will be omitted.

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

1 is a process diagram of a low temperature fluid lithography method according to the present invention.

As shown in FIG. 1, a low temperature fluid lithography method according to the present invention comprises the steps of (a) flowing a photocurable fluid 1 into a microchannel 10 (S100), and (b) a microchannel 10 Irradiating light (3) to the step (S200) of synthesizing the fine particles (5), wherein steps (a) and (b) are performed at a synthesis temperature of low temperature, which is relatively lower than room temperature. .

The present invention relates to a method for synthesizing microparticles (5) using fluid lithography. Herein, the particles 5 to be synthesized are formed of a polymer in which the photocurable monomers constituting the particle precursor are polymerized into a mesh structure. For example, the particles 5 may be hydrogel particles. However, the fine particles 5 to be synthesized are not necessarily limited to the hydrogel particles, and various particles may be synthesized according to the particle precursor.

These fine particles 5 are synthesized according to the following method.

First, the photocurable fluid 1 flows into the microchannel 10 (S100). According to the present invention, a microfluidic chip having a microchannel 10 is used, and the photocurable fluid 1 can be flowed by injecting the photocurable fluid 1 into an injection hole communicating with the microchannel 10. .

Here, the photocurable fluid 1 may include a photocurable monomer and a photoinitiator as precursors of the microparticles 5 to be synthesized. As an example of the photocurable monomer, polyethylene glycol diacrylate (PEGDA) may be used. However, the present invention is not limited thereto, and in addition to hydrophilic PEGDA, trimethylolpropane ethoxylate triacrylate (TMPETA), which is a hydrophobic substance, may be used. At this time, since the photocurable fluid 1 should have fluidity, the photocurable monomer and the photoinitiator may exist in an aqueous solution mixed with water. On the other hand, the photocurable monomers may be included in a plurality of different types.

Furthermore, in order to impart specific functions to the synthesized microparticles 5, the photocurable fluid 1 may further contain a functional substance. Here, the functional material may further include at least one or more of a material copolymerized with the photocurable monomer, and nanoparticles. In this case, the copolymer material may be any one or more selected from the group consisting of a fluorescent material, a DNA probe (probe), an antibody, a virus, a protein, and a metal compound as a material to be copolymerized in the particle network. On the other hand, nanoparticles are particles that are mounted between the particle network, for example, may include any one or more selected from the group consisting of nano-fluorescent particles, nano-magnetic particles, proteins, nano-metal particles, nano-chemical particles, and cells. Can be. However, the copolymer and the nanoparticles are not necessarily limited to the above-described types, and may be used without limitation as long as they are copolymerized on the mesh of the particles 5 or can be mounted.

Meanwhile, the microchannel 10 may be formed of PDMS (polydimethylsiloxane) so that the synthesized microparticles 5 flow without being adsorbed on the inner wall of the channel 10. Since PDMS has excellent oxygen permeability, oxygen passing through the channel 10 wall consumes radicals generated by the photoinitiator, leading to a reaction in which the polymerization of the polymer is stopped. At this time, a lesser oligomer-like lubrication layer is formed on the inner wall of the PDMS channel 10 so that the microparticles 5 do not stick to the wall of the channel 10.

In addition, an inlet for injecting the photocurable fluid 1 is connected to one end of the microchannel 10, and an outlet for injecting the synthesized fine particles 5 is connected to the other end of the microchannel 10. In this case, the inlet or outlet may be provided with a plurality of branched ends of the microchannel 10.

When the photocurable fluid 1 flows through the thus formed microchannel 10, the light 3 is irradiated toward the microchannel 10 using the light source 20 (S200). In this case, the light 3 used may be ultraviolet (UV) light.

On the other hand, in order to shape the fine particles 5 into a predetermined shape, the mask 30 can be used. In the mask 30, a pattern formed in the direction in which the light 3 is irradiated is formed. Therefore, when the light 3 passes through the mask 30 and irradiates the photocurable fluid 1, the fine particles 5 having a cross section corresponding to the puncturing pattern can be synthesized.

In addition, a lens 40 may be disposed between the mask 30 and the microchannel 10. The lens 40 guides the light 3 passing through the mask 30 to be focused toward the microchannel 10 without being emitted.

Here, the microparticles 5 to be synthesized may be formed in a polyhedral shape, etc. In this case, in the polyhedral microparticles 5, the microparticles have a height of 1 to 200 μm in cross section and a width of 1 to 500 in cross section. May be μm. However, the shape, height, and width of the fine particles 5 are not necessarily limited thereto.

In the present invention, the above-described processes (S100, S200) are carried out at a low temperature synthesis temperature is relatively lower than the room temperature. Specifically, the synthesis temperature may be 0 ~ 15 ℃. Since 0 ° C. is a point of water contained in the photocurable fluid 1 as the fine particle 5 precursor, condensation occurs severely below 0 ° C., so that the fluidity of the photocurable fluid 1 cannot be secured. Moreover, when it exceeds 15 degreeC, the completeness of the geometric shape of the microparticles | fine-particles 5 will fall, and the loading rate of the functional substance contained in the microparticles | fine-particles 5 will become low. In other words, when the synthesis temperature is in the range of 0 to 15 ° C, it has a critical significance in which the change in technical characteristics is soaring around the lower limit and the upper limit, and has a significant effect (technical significance) in the whole range. This will be described later in detail through the examples.

In the following, a system capable of implementing the aforementioned low temperature fluid lithography method is described. Details that overlap with the above-described parts of the contents will be omitted or simply described.

2 is a perspective view schematically showing a low temperature fluid lithography system according to a first embodiment of the present invention.

As shown in FIG. 2, the low temperature fluid lithography system according to the first embodiment of the present invention includes a microfluidic chip 100 and a microchannel having a microchannel 110 through which a photocurable fluid 1 flows. The light source 200 for irradiating the light 3 to the light source 110 to synthesize the fine particles 5 and the cooling unit 300 for maintaining the temperature in the microchannel 110 at a low temperature synthesis temperature that is relatively lower than room temperature. ).

The present invention relates to a system for realizing low temperature fluid lithography, comprising a microfluidic chip (100), a light source (200), and a cooling unit (300).

The microfluidic chip 100 includes a microchannel 110, an inlet 120 communicating with one end of the microchannel 110, and an outlet 130 communicating with the other end of the microchannel 110. Here, the microfluidic chip 100 is provided with a concave recessed recess in the lower surface of the upper plate 140, the lower plate 150 is coupled to the lower surface of the upper plate 140 to form a micro channel (110). Can be. In this case, the upper plate 140 and the lower plate 150 may be made of PDMS, the glass plate 160 may be disposed on the lower surface of the lower plate 150. However, the shape and configuration of the microfluidic chip 100 are not necessarily limited thereto, and the microfluidic chip 100 that can be used for a normal fluid lithography may be used without particular limitation.

The light source 200 is a device for irradiating the light 3 to the microchannel 110, and the irradiated light 3 may be ultraviolet rays. Here, a mask 400 for determining the shape of the microparticles 5 may be disposed between the microchannel 110 and the light source 200, and a lens (between the mask 400 and the microchannel 110). 500 may be disposed.

The cooling unit 300 is a device that maintains the temperature in the micro channel 110 relatively lower than room temperature. In the present invention, the fluid lithography is performed at a lower temperature than room temperature, where the cooling unit 300 maintains the synthesis temperature of the fine particles 5 at a low temperature. At this time, the synthesis temperature may be maintained at 0 ~ 15 ℃ as described above.

As long as the cooling unit 300 can maintain the synthesis temperature at a low temperature, there is no particular limitation on the specific configuration, but some embodiments will be described below.

The cooling unit 300a according to the first embodiment of the present invention may include a heat conduction plate 310a and a cooling device 310b. Here, the thermal conductive plate 310a contacts the outer surface of the microfluidic chip 100 to cool the high temperature microfluidic chip 100 to low temperature by thermal conduction. At this time, the high temperature and low temperature means a relative temperature, the heat conduction plate 310a is maintained lower than the temperature of the microfluidic chip 100, where the cooling device 330a cools the heat conduction plate 310a. Specifically, a channel through which cooling fluid flows is provided in the heat conduction plate 310a, and the cooling device 330a may be operated by injecting cooling fluid into the channel. At this time, the cooling fluid is a fluid of low temperature, for example, cooling water, cooling oil, etc. can be used (it is the same below).

3 is a perspective view schematically showing a low temperature fluid lithography system according to a second embodiment of the present invention, wherein the cooling unit 300b according to the second embodiment of the present invention includes a cooling fluid 310b and a cooling fluid circulation channel. 330b. Here, the cooling fluid circulation channel 330b is formed to allow the cooling fluid 310b to flow inside the microfluidic chip 100, so that the cooling fluid 310b circulates around the microchannel 110 and has a predetermined synthesis temperature. Keep it. Meanwhile, the cooling fluid circulation channel 330b is connected to a thermostat (not shown) that receives the cooling fluid 310b and maintains the cooling fluid 310b at a constant temperature, thereby continuously circulating the cooling fluid 310b.

4 is a perspective view schematically showing a low temperature fluid lithography system according to a third embodiment of the present invention.

As shown in FIG. 4, the cooling unit 300c according to the third embodiment of the present invention may include a cooling fluid 310c and a cooling fluid circulation tube 330c. Here, the cooling fluid 310c flows along the inside of the cooling fluid circulation tube 330c, and the cooling fluid circulation tube 330c is disposed in contact with the outer surface of the microfluidic chip 100. Therefore, while the cooling fluid 310c flows along the cooling fluid circulation tube 330c, the outer surface of the microfluidic chip 100 is cooled to maintain a synthesis temperature. Meanwhile, in order to continuously circulate the cooling fluid 310c, the cooling fluid circulation tube 330c may be connected to a thermostat (not shown) that receives the cooling fluid 310c and maintains the temperature at a predetermined temperature.

5 is a perspective view schematically showing a low temperature fluid lithography system according to a fourth embodiment of the present invention.

As shown in FIG. 5, the cooling unit 300d according to the fourth embodiment of the present invention may include a container unit 310d and a coolant 330d. Here, the container portion 310d has a double wall structure in which the inner wall and the outer wall are spaced apart from each other by a predetermined interval, and the microfluidic chip 100 is disposed in a space surrounded by the inner wall. The coolant 330d is a material having a low temperature and is disposed in a space between the inner wall and the outer wall of the container portion 310d in a gas, liquid, or solid state to maintain the synthesis temperature at a low temperature.

Meanwhile, the cooling units 300a to 300d according to the first to fourth embodiments described above may include a thermometer or a temperature sensor (not shown) to measure the synthesis temperature in real time, and furthermore, a temperature control device (not shown). To maintain a constant synthesis temperature during the entire fluid lithography process.

However, since the cooling unit 300 according to the present invention is not necessarily limited to the first to fourth embodiments, the cooling unit 300 may be operated by lowering the temperature of the indoor space in which the microfluidic chip 100 is disposed.

Overall, according to the present invention, the fluid lithography is performed at a low temperature, thereby improving the degree of polymerization of the particles so that the particles have a more complete geometric shape, and the functional materials are copolymerized or loaded at a high rate to ensure the improved functionality of the particles. Can be. In addition, when the biomolecule detection probe is copolymerized to the particles at a low temperature, the detection sensitivity is increased and the detection limit is lowered, thereby saving the starting materials and increasing the economic efficiency, as well as improving the performance of the functional particles.

The above effect according to the present invention is demonstrated by the following specific examples.

Example 1 Microfluidic Chip Fabrication

In this embodiment, a microfluidic chip is manufactured. First, in order to design the microchannels, the channel design is embossed on the SU-8 master wafer, and PDMS is poured on the master wafer and cured at 70 ° C. for at least 6 hours. After removing the cured PDMS and cut into a predetermined shape, a hole is formed to form an inlet and an outlet to prepare a top plate. Next, PDMS is lightly coated on the glass plate and partially cured in an oven for 20 minutes to complete the lower plate. Finally, the microfluidic chip was prepared by attaching the upper and lower plates and completely curing in an oven. Here, the height of the microchannel is 50 μm.

Example 2: Microparticle Synthesis at Low Temperature

In this embodiment, the microparticles were synthesized at low temperature using the microfluidic chip prepared in Example 1. Here, the precursor is 40% (v / v) polyethylene glycol diacrylate (PEGDA) 700, 20% (v / v) polyethylene glycol (PEG) 200, 35% (v / v) water, 5% (v / v) photoinitiator (Darocur 1173) was prepared by mixing. The precursor was injected into the microchannel through the inlet of the microfluidic chip, and the microparticles were synthesized by exposure to ultraviolet rays. At this time, the intensity of the ultraviolet ray was fixed at 165 ㎽ / ㎠, exposure time 75 ㎳, the synthesis temperature was maintained at 10 ℃.

Comparative Example 1 Microparticle Synthesis at High Temperature

Each synthesis temperature was set to 23 ℃, 35 ℃, the remaining conditions were the same as in Example 2 to synthesize the fine particles.

Evaluation Example 1: Comparison of Particle Size

After washing the microparticles synthesized in Example 2 and Comparative Example 1 with water containing 0.05% of Tween 20, the size was measured. 6 is a photograph of microparticles synthesized by a fluid lithography method.

For reference, the average height of the particles synthesized at a low temperature (10 ℃) according to Example 2 was measured as 48 ㎛ width 37 ㎛. On the contrary, the average height of the particles synthesized at room temperature (23 ° C.) of Comparative Example 1 was measured to be 44 μm in width.

As a result, the height and width of the particles synthesized at low temperature were improved by about 10% compared to those synthesized at room temperature. In addition, particles synthesized at low temperatures had sharper edges. Thus, according to the low temperature fluid lithography according to the invention, it can be seen that the microparticles are synthesized into a more complete geometric shape.

Example 3: Microparticle Synthesis Containing Copolymer at Low Temperature

Under the same conditions as in Example 2, microparticles were synthesized by adding Rhodamine B Methacrylate as a fluorescent functional material to the precursor. At this time, the precursor is 40% (v / v) polyethylene glycol diacrylate (PEGDA) 700, 20% (v / v) polyethylene glycol (PEG) 200, 34% (v / v) water, 5% (v / v) photoinitiator (Darocur 1173) Consists of 1% (v / v) Rhodamine B Methacrylate. Here, the cross-sectional shape of the particles was synthesized in a circle and coded.

Comparative Example 2: Synthesis of Fine Particles Containing Copolymer at High Temperature

Each synthesis temperature was set to 23 ° C, 35 ° C, and the remaining conditions were the same as in Example 3 to synthesize fine particles. At this time, the cross-sectional shape of the particles synthesized at 23 ° C. was rectangular, and the cross-sectional shapes of the particles synthesized at 35 ° C. were coded into hexagons.

Evaluation Example 2: Comparison of Copolymer Content

The microparticles synthesized in Example 3 and Comparative Example 2 were washed with water containing 0.05% of Tween 20, and then measured by fluorescence intensity. In FIG. 7, images and graphs showing the fluorescence intensity of each microparticle are shown.

As a result, the average fluorescence intensity of the particles synthesized in Example 3 was about 15% higher than the average fluorescence intensity of the particles synthesized in Comparative Example 2. This means that more materials are copolymerized during low temperature synthesis.

Example 4 Microparticle Synthesis Containing Nanoparticles at Low Temperature

Each synthesis temperature was set at 0 ° C., 10 ° C., and 15 ° C., and the rest of the conditions were the same as in Example 2, whereby Fluorescein Isothiocyanate-Bovine Serum Albumin (FITC-BSA), a fluorescent nanoparticle mounted between the meshes of particles, In addition to the precursor, microparticles were synthesized. At this time, the precursor is 40% (v / v) polyethylene glycol diacrylate (PEGDA) 700, 20% (v / v) polyethylene glycol (PEG) 200, 30% (v / v) water, 5% (v / v) photoinitiator (Darocur 1173) It consists of 5% (v / v) FITC-BSA. Here, the cross-sectional shape of the microparticles synthesized at 0 ° C was circular, the cross-sectional shape of the microparticles synthesized at 10 ° C was hexagonal, and the cross-sectional shape of the microparticles synthesized at 15 ° C was triangulated.

Comparative Example 3: Synthesis of Microparticles Containing Nanoparticles at High Temperature

Each synthesis temperature was set to 23 ° C., and the fine particles were synthesized in the same manner as in Example 4 above. At this time, the cross-sectional shape of the particles synthesized at 23 ° C was coded into a rectangle.

Evaluation Example 3: Comparison of Nanoparticle Content

After washing the microparticles synthesized in Example 4 and Comparative Example 3 with water containing 0.05% of Tween 20, in order to determine the amount of the nanoparticles contained, the respective fluorescence intensity was measured and compared. 8 shows an image and a graph showing fluorescence intensity of each microparticle.

As a result, since the average fluorescence intensity of the particles synthesized in Example 4 is about 17% higher than the average fluorescence intensity of the particles synthesized in Comparative Example 3, it was confirmed that more nanoparticles were loaded during low temperature synthesis.

Example 5 Microparticle Synthesis Containing DNA Probe at Low Temperature

In order to demonstrate the application effect in the field of micro RNA biomolecule detection, microparticles were synthesized under the same conditions as in Example 2, including a precursor containing a DNA probe copolymerized in the mesh of the particles. Here, the precursor used consisted of a DNA probe (1 mM) complementary to the 90% (v / v) particle constituent precursor and the 10% (v / v) target microRNA, wherein the particle constituent precursor was 20% (v / v). ) polyethylene glycol diacrylate (PEGDA) 700, 40% (v / v) polyethylene glycol (PEG) 200, 35% (v / v) 3% Tris EDTA, 5% (v / v) photoinitiator (Darocur 1173) . Here, the cross-sectional shape of the microparticles was coded in a circle.

Comparative Example 4: Synthesis of Microparticles Containing DNA Probe at High Temperature

Each synthesis temperature was set to 23 ° C and 35 ° C, and the remaining conditions were the same as in Example 5 to synthesize fine particles. At this time, the cross-sectional shape of the particles synthesized at 23 ° C. was rectangular, and the cross-sectional shape of the particles synthesized at 35 ° C. was coded into hexagons.

Evaluation Example 4: Comparison of Detection Sensitivity

After recovering and washing the particles synthesized in Example 5 and Comparative Example 4, and reacting the target microRNA of artificial concentration, and measuring the sensitivity by converting the microRNA reacted with the particles to the fluorescence intensity through a series of fluorescence detection procedures It was. In FIG. 9, an image and a graph showing fluorescence intensity of each microparticle are shown.

As a result, it was confirmed that the average fluorescence intensity of the particles synthesized in Example 5 was about 20% higher than the average fluorescence intensity of the particles synthesized in Comparative Example 4. This indicates that more probes are copolymerized at low temperature to improve detection sensitivity.

Example 6: Microparticle Synthesis Using TMPETA at Low Temperature

In the precursor of Example 4, PEGDA was replaced with Trimethylolpropane ethoxylate triacrylate (TMPETA) to synthesize microparticles at 0 ° C and 10 ° C synthesis temperatures, respectively. Here, the cross-sectional shape of the microparticles synthesized at 0 ° C was circular, and the cross-sectional shape of the microparticles synthesized at 10 ° C was coded into hexagons.

Comparative Example 5: Microparticle Synthesis Using TMPETA at Low Temperature

The synthesis temperature was 23 ° C., and the remaining conditions were the same as those in Example 6, to synthesize fine particles having a rectangular cross-sectional shape.

Evaluation Example 5: Comparison of Nanoparticle Content

In order to confirm whether the same effect as PEGDA is expressed even when a material other than PEGDA, which is used most often as a material for forming a mesh of particles in a fluid lithography process, the microparticles synthesized in Examples 6 and 5 were used After washing with 0.05% water containing Tween 20, the fluorescence intensity was measured and the amount of nanoparticles contained was compared. In FIG. 10, an image and a graph showing fluorescence intensity of each microparticle are shown.

Comparing FIG. 8 with FIG. 10, it can be seen that the trend is quite similar to that of PEGDA particles. Thus, it can be seen that the effect of low temperature fluid lithography according to the present invention is applied not only to PEGDA particles but also to particles composed of other polymers.

Although the present invention has been described in detail through specific examples, it is intended to describe the present invention in detail, and the present invention is not limited thereto, and should be understood by those skilled in the art within the technical spirit of the present invention. It is obvious that modifications and improvements are possible.

All modifications and variations of the present invention fall within the scope of the present invention, and the specific scope of protection of the present invention will be apparent from the appended claims.

1: photocurable fluid 3: light
5: microparticles 10, 110: microchannel
20, 200: light source 30, 400: mask
40, 500: Lens 100: Microfluidic chip
120: inlet 130: outlet
140: top plate 150: bottom plate
160: glass plate 300, 300a, 300b, 300c, 300d: cooling unit

Claims (12)

(a) flowing the photocurable fluid into the microchannel; And
(b) irradiating light onto the microchannels to synthesize fine particles;
Including,
The step (a) and the step (b) are carried out at a synthesis temperature of 0 ~ 15 ℃ low temperature fluid lithography method.
delete The method according to claim 1,
The photocurable fluid is
A photocurable monomer, and a photoinitiator,
A low temperature fluid lithography method further comprising at least one of a copolymer, and nanoparticles.
The method according to claim 1,
The fine particles
The microparticles have a polyhedral shape having a cross-sectional height of 1 to 200 μm and a cross-sectional width of 1 to 500 μm.
The method according to claim 3,
The copolymer material
A low temperature fluid lithography method comprising at least one selected from the group consisting of fluorescent materials, DNA probes, antibodies, viruses, proteins, and metal compounds.
The method according to claim 3,
The nanoparticles
A low temperature fluid lithography method comprising at least one selected from the group consisting of nanofluorescent particles, nanomagnetic particles, proteins, nanometal particles, nanochemical particles, and cells.
A microfluidic chip having a microchannel through which a photocurable fluid flows;
A light source for irradiating light to the microchannels to synthesize fine particles; And
Cooling unit for maintaining the temperature in the micro-channel to a synthesis temperature of 0 ~ 15 ℃;
Low temperature fluid lithography system comprising a.
The method according to claim 7,
The cooling unit
A thermal conductive plate in contact with an outer surface of the microfluidic chip; And
A cooling device for cooling the heat conducting plate;
Low temperature fluid lithography system comprising a.
The method according to claim 7,
The cooling unit
Cooling fluid; And
A cooling fluid circulation channel formed inside the microfluidic chip so that the cooling fluid flows;
Low temperature fluid lithography system comprising a.
The method according to claim 7,
The cooling unit
Cooling fluid; And
A cooling fluid circulation tube in contact with an outer surface of the microfluidic chip and formed such that the cooling fluid flows;
Low temperature fluid lithography system comprising a.
The method according to claim 7,
The cooling unit
An inner wall and an outer wall formed of a double wall structure spaced apart from each other, and a container part in which the microfluidic chip is disposed in a space surrounded by the inner wall; And
A coolant disposed between the inner wall and the outer wall;
Low temperature fluid lithography system comprising a.
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