KR20140114552A - Apparatus for microalgae screening with multi-layered microfluidic channel and method for screening microalgae using the same - Google Patents

Abstract

The present invention relates to a device for screening microalgae using a multi-layer microfluidic channel. The device comprises a first layer (110), a second layer (120), and a third layer (130) which are stacked vertically. 1-1 feeding units (111a, 111b) and 1-2 feeding units (113a, 113b) are formed to penetrate through the first layer. 2-1 feeding holes (121a, 121b), a second microchannel unit (124), and a 2-3 feeding unit (125) which are sequentially connected with the 1-1 feeding units (111a, 111b) are formed in the second layer. 3-1 feeding units (133a, 133b), a third microchannel unit (134), and a chamber unit (136) are formed in the third layer. In the second microchannel unit and third microchannel unit, a step in which one channel is branched and is simultaneously coupled with another neighboring channel is repeated. Through the device according to the present invention, various process variables can be simultaneously tested in one chip consisting of a multi-layer microfluidic channel in which a fluid flows. Accordingly, in the present invention, an optimal condition for the growth of microalgae can be rapidly screened. Moreover, because the microfluidic channel is used, problems such as device expansion can be effectively solved.

Description

TECHNICAL FIELD The present invention relates to a micro-algae screening apparatus using a multi-layer microfluidic channel, and a micro-algae screening method using the same. [0002] The present invention relates to an apparatus and method for screening micro-

The present invention relates to a micro-algae screening apparatus using a microfluidic channel and an apparatus for producing microalgae using the same. More particularly, the present invention relates to a micro-algae screening apparatus using a microfluidic channel, The present invention relates to a micro-algae screening apparatus using a multi-layer microfluidic channel and a method of screening micro-algae using the same.

Microalgae (microalgae) are photosynthetic single-celled organisms with photosynthetic pigments and are currently attracting attention as a new source of supply in the fields of energy, chemistry and environment.

Cultivation and growth of these microalgae are affected by various external conditions. For example, the growth rate of each microalgae varies depending on the carbon source and pH conditions. If there are a plurality of kinds of microalgae for growth analysis, there is a problem that the number of factor combinations for growth analysis is further increased as shown in the following equation.

Number of factor combinations for analysis of microalgae growth = type of microalgae × carbon source × temperature × light source × amount of nutrients × pH condition etc.

The prior art finds optimal process variables in such a manner that only one of the various process parameters is repeatedly changed. In this case, a large amount of expensive reagent is repeatedly used, a repeated labor-intensive experimental process is performed, , The screening time itself becomes longer, the microalgae culture space becomes excessively large, and the price analysis efficiency becomes lower.

Therefore, a more economical and efficient screening means was required.

An object of the present invention is to provide an integrated micro-algae screening apparatus that is miniaturized in a more economical manner.

The present invention is characterized in that it comprises a first layer 110, a second layer 120 and a third layer 130 which are stacked one above the other and the first layer is provided with first 1-1 inlet ports 111a and 111b, 2 injection ports 113a and 113b are formed in the first layer and the second layer is provided with second-1 injection ports 121a and 121b that sequentially communicate with the first-first injection ports 111a and 111b, The channel portion 124 and the second to third injection ports 125 are formed in the first layer 110 and the third layer is formed in the third layer, 1 inlet 133a and 133b and a third microchannel 134 and a chamber 136. The second microchannel and the third microchannel are separated from each other by one channel, The micro-algae screening apparatus using the multi-layer microfluidic channel is provided.

The second layer is provided with a second 2 inlet 123a communicating with the first 1-2 inlet 113a of the first layer 110 and the 3-1 inlet 133a of the third layer 133b, And 123b are further formed.

The 2-1 inlet ports 121a and 121b and the second microchannel section 124 are formed on the surface of the second layer and the 2-3 inlet 125 is formed through the second layer .

And the second to third injection ports 125 of the second layer are formed to communicate with the chamber part 135 of the third layer.

In the second microchannel portion and the third microchannel portion, a sequential concentration gradient of the material flowing in the channel is formed while branching and joining the channels.

The chamber portion 136 is composed of a groove 136a formed in a long surface and a C-shaped capturing portion 136b arranged in a plurality of spaces at a predetermined interval in the longitudinal direction of the groove.

The material injected into the first and second inlets 113a and 113b of the first layer 110 is microalgae or nutrients.

An outlet is provided through the first, second and third layers to discharge material within the chamber portion.

The third layer further includes temperature adjusting means for adjusting a temperature of the chamber below the chamber portion.

(200) for injecting the material into the 1-1 inlet or the 1-2 inlet of the first layer.

The present invention also provides a method for screening microalgae using a microalgae screening apparatus comprising a first layer 110, a second layer 120 and a third layer 130 stacked in order in the vertical direction, A first step of injecting a microalgae into the first step; A second step of passing the microalgae through the second layer and being accommodated in a plurality of chamber parts of the third layer; A third step of injecting a first nutrient into the first layer; A fourth step of injecting the first nutrient of the third step into the chamber part of the third layer after a concentration gradient is formed in the channel formed on the surface of the second layer; A fifth step of injecting a second nutrient into the first layer; And a sixth step of injecting the second nutrient of the fifth step into the chamber part of the third layer after a concentration gradient is formed in the channel formed on the surface of the third layer. .

In the fourth through sixth steps, the concentration gradient of the substance is repeated by repeating the process of joining one channel to the neighboring channel while branching. And heating the lower portion of the chamber portion to adjust the temperature.

Through the screening apparatus according to the present invention, it is possible to simultaneously test various process parameters in a single chip composed of a multi-layered microfluidic channel in fluid communication. Therefore, in the present invention, optimal conditions for microalgae growth can be screened in a short period of time, and since the microfluidic channel is used, problems such as device enlargement can be effectively solved.

1 is an exploded perspective view of a microalgae screening apparatus using a multi-layer microfluidic channel according to an embodiment of the present invention,
FIG. 2 is a view of a first layer of the multi-layer microalgae screening apparatus shown in FIG. 1,
FIG. 3 is a view of the second layer of the multi-layer microalga screening apparatus shown in FIG. 1,
FIG. 4A is a view of the third layer of the multi-layer microalga screening apparatus shown in FIG. 1,
4B is a SEM photograph of the actual chamber formed in the third layer,
FIG. 4c is a view showing that cells are being cultured in the chamber part of the third layer,
5 to 6 are views showing a temperature control means provided under the multi-layer microalgae screening apparatus shown in FIG. 1,
FIG. 7 is a view illustrating injection of microalgae into a microalgae screening apparatus using a multi-layer microfluidic channel according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail with reference to the drawings. The following embodiments are provided by way of example so that those skilled in the art can fully understand the spirit of the present invention. Therefore, the present invention is not limited to the embodiments described below, but may be embodied in other forms. In the drawings, the width, length, thickness, etc. of components may be exaggerated for convenience. Like reference numerals designate like elements throughout the specification. In addition, abbreviations displayed throughout this specification should be interpreted to the extent that they are known and used in the art unless otherwise indicated herein.

It is to be understood that the terms "comprises", "comprising", or "having" as used in the foregoing description mean that the constituent element can be implanted unless specifically stated to the contrary, But should be construed as further including other elements. All terms, including technical and scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, unless otherwise defined. Commonly used terms, such as predefined terms, should be interpreted to be consistent with the contextual meanings of the related art, and are not to be construed as ideal or overly formal, unless expressly defined to the contrary.

The foregoing description is merely illustrative of the technical idea of the present invention and various changes and modifications may be made by those skilled in the art without departing from the essential characteristics of the present invention. Therefore, the embodiments disclosed in the present invention are intended to illustrate rather than limit the scope of the present invention, and the scope of the technical idea of the present invention is not limited by these embodiments. The scope of protection of the present invention should be construed according to the following claims, and all technical ideas falling within the scope of the same shall be construed as falling within the scope of the present invention.

1 is an exploded perspective view of a microalgae screening apparatus using a multi-layer microfluidic channel according to an embodiment of the present invention.

A micro-algae screening apparatus using a multi-layered microfluidic channel according to the present invention (hereinafter referred to as a screening apparatus for the sake of convenience) comprises a plurality of layers stacked vertically. Also, the term " microfluid channel " or " microchannel in microchannel " as used herein includes any minute unit (micro-nano unit) and is not limited to a particular numerical range.

1, the screening apparatus 100 shown in FIG. 1 includes three layers of a first layer 110, a second layer 120, and a third layer 130 from top to bottom , More layers may be added as needed. Preferably, the first layer 110 and the third layer 130 comprise a PDMS layer and the second layer comprises a glass layer.

In each of the first to third layers, micro-algae and microfluidic channels are formed, which are the channels through which the micro-algae and the carbon source and nutrients necessary for the growth of the micro-algae and the pH,

Figs. 2 to 4A are views of respective layers of the multi-layer microalgae screening apparatus. The first layer 110 is formed as an uppermost layer of the screening apparatus and is formed with upper and lower first inlet ports 111a and 111b adjacent to the upper side and first and second inlet ports 113a and 113b Respectively. A first outlet 118, which is a passage through which the injected substance (solution, nutrient or microalgae, hereinafter referred to as "substance") or microalgae is discharged to the outside, is formed through the first layer.

The solution or the like injected into the first 1-1 injection ports 111a and 111b of the first layer 110 passes through the first layer and then passes through the second layer 1 of the second layer 120, (121a, 121b).

Referring to FIG. 3, on the surface of the second layer 120, channels through which the fluids flowing from the 2-1 inlet 121a and the 2-1 inlet 121b are formed. The second 2-1 channels 122a and 122b are communicated with the 2-1 inlet 121a and 121b and the second microchannel 124 is provided below the 2-1th channel 122a and 122b. do. The second micro-channel unit is a place where the second-1 channels 122a and 122b continue to branch and merge with neighboring channels. That is, the second microchannel unit continuously repeats the process of joining with its neighboring channel while the channel of the second microchannel is branched, Are mixed with each other as they pass through this area, and as a result, the concentrations are mixed with successively different gradients.

That is, when a material A is injected into only the left inlet 121a from the second inlet 1 (121a, 121b), the material A passes through the second microchannel 124, As a result, the concentration gradient of the A material gradually decreases from the left to the right of the second microchannel portion 124 as shown in FIG. 3. The second micro-channel part 124 is formed at the downstream side with a second through-hole injection port 125 passing through the second micro-channel part as much as the channel number. The material flowing into the second through- The concentration gradient described above is obtained.

 The material that has passed through the second to third injection port 125 flows into the via hole 135 formed in the third layer 130, which will be described below.

In addition, second and third inlet ports 123a and 123b are formed in the center of the second layer 120 so as to communicate with the first and second inlet ports 113a and 113b of the first layer 110, do.

Referring to FIG. 4A, on the surface of the third layer 130, channels for flowing the fluid introduced into the 3-1 inlet ports 133a and 133b and the 3-1 inlet port are formed.

The material injected into the first and second inlets 113a and 113b of the first layer 110 passes through the second inlet 2 123a and the inlet 123b of the second layer 120, , 133b.

Similarly to the second layer, a third microchannel 134 is provided on the downstream side of the 3-1 inlet ports 133a and 133b. The third microchannel portion is a place where the channels in the downstream direction of the 3-1 inlet ports 133a and 133b continue to branch and merge. That is, the third micro-channel unit continuously repeats the process of joining its neighboring channels while its channel is branched. The materials introduced from the 3-1 inlet ports 133a and 133b are mixed with each other as they pass through the third microchannel portion, and are sequentially mixed with different concentration gradients. That is, when a material B is injected into the left inlet 133a from the third inlet 1 (133a, 133b), the material B passes through the third microchannel 134, As a result, the concentration gradient of the B material gradually decreases from the left to the right of the plurality of third microchannel portions, as shown in FIG. 4.

A plurality of via holes 135 through which the material passing through the third microchannel 134 flows are formed on the downstream side of the third microchannel 134.

At the downstream of the via hole 135, a chamber part 136 is formed in which micro-algae are captured and a carbon source of various concentrations is injected. 4A is an enlarged view of the chamber part 136. The chamber part 136 includes grooves 136a formed on the surface thereof so as to be a passage through which the carbon source flows, And a C-shaped capturing portion 136b arranged in a plurality of the openings. A portion of the groove 136a where the capturing portion 136b is disposed rounds the side of the groove in a circular shape along the shape of the capturing portion so that the size of the capturing portion is received in the groove. The fine alga that passes through the via hole 135 is captured by the C-shaped capturing portion 136b of the chamber portion 136, and a carbon source or the like is supplied in this state.

FIG. 4B is an enlarged photograph of an actual SEM of the chamber portion, and FIG. 4C is an actual view of cells being cultured in the chamber portion.

5 to 6 are views showing a temperature adjusting unit provided below the screening apparatus according to the present invention. 5 shows a state in which the light 310 is irradiated and heated at the lower ends of the plurality of via holes 135 of the third layer 130. FIG. And the temperature of the via hole is adjusted by heating the fine heater 320 by heating. The via hole 135 may be adjusted to the same temperature, or a temperature gradient may be given to observe microalgal growth with temperature change. That is, the heating amount can be adjusted from left to right in FIGS. Although the figure shows heating the via hole 135, it may be heated to control the temperature of the chamber part 136, not the via hole.

FIG. 7 is a view illustrating injection of microalgae into a microalgae screening apparatus using a multi-layer microfluidic channel according to an embodiment of the present invention. The operation of the screening apparatus according to the present invention described above will be described below with reference to FIG.

In the screening apparatus according to the present invention, microalgae requiring growth analysis are injected and a substance such as carbon source, which is a factor of growth analysis, is injected. As the carbon source, glucose (glucose), sodium acetate, etc. can be used. The means for injecting is not limited to a specific means, and various means are possible. In FIG. 7, the injection means 200 in the form of a syringe is exemplarily shown. The injected material is injected into the cylindrical body portions 210a and 210b and then the pressurizing means 220 is pressed into the injection passages 240a and 240b, 240b are injected into the screening apparatus.

In this embodiment, the microalgae are injected into the 1-2 th injection ports 113a and 113b of the first layer 110 of the screening apparatus 100 of the present invention. Thereafter, the microalgae are injected into the 3-1 inlet ports 133a and 133b through the 2-2 inlet ports 123a and 123b of the second layer 120, And is accommodated in the chamber portion 136 via the channel portion 134 and the via hole 135.

When the microalgae of the same concentration are injected into the body portions 210a and 210b of the injection means, the microalgae of the same concentration will be accommodated in the chamber portion 136 as well. The principle that a substance contained in the chamber portion passes through the microchannel and a concentration gradient is generated has been described above and will not be described here.

In the state in which the microalgae are accommodated in the chamber part 136, a substance (for example, a carbon source) which is a factor for the growth analysis of microalgae is injected in the following manner.

The carbon source was injected into the 1-1 inlet (111a, 111b) of the first layer (110) using a separate injection means. In order to observe the effect of the microalgae on the concentration gradient, The carbon source is injected into only one of the first and second catalyst layers 111a and 111b, and the other material is supplied with a concentration gradient (for example, water is introduced).

The carbon source flows through the first layer 110 through the 1-1 inlet ports 111a and 111b and then into the 2-1 inlet ports 121a and 121b of the second layer 120. [ The carbon source is mixed with the concentration gradient while flowing from the 2-1 inlet 121a and the 121b to the 2-1 channel 122a and 122b and the second microchannel 124, 3 inlet (125).

The second to third injection ports 125 are formed through the second layer 120 and are formed to directly communicate with the upper portion of the chamber 135 of the third layer 130. Therefore, the carbon source introduced into the second to third injection port 125 flows into the via hole 135, and then is supplied to the microwave oven accommodated in the chamber part 136.

After one carbon source is injected into the chamber portion with a concentration gradient through the 1-1 inlet ports 111a and 111b of the first layer 110, The first and second inlets 113a and 113b may be injected into the chamber portion 136 so as to be supplied with a concentration gradient. The nutrients injected into the first and second inlets 113a and 113b of the first layer 110 are injected into the chamber portion 136 of the third layer 130 through the second layer 120 while forming a concentration gradient The principle has already been described above, so it is omitted here.

In this way, micro-algae can be injected into the chamber and the two carbon sources can be supplied in a concentration gradient, thereby enabling them to efficiently analyze the influence of micro-algae growth rate.

When the analysis necessary for the growth analysis is completed, the material contained in the chamber part 136 can be discharged through the discharging means of the screening device. That is, the first outlet 118 of the first layer, the second outlet 128 of the second layer 120, and the third outlet 138 of the third layer 130 are formed to communicate with each other, So that the material contained in the chamber 135 can be discharged to the outside.

Although the micro-algae screening apparatus using the multilayer microfluidic channels stacked in three layers has been described above, it is not necessarily limited to three layers, and a screening apparatus in which more PDMS layers are stacked may be used as needed .

100: Micro-algae screening device using multilayer microfluidic channel
110: first layer 111a, 111b:
113a, 113b: first-second inlet port 118: first outlet port
120: second layer 121a, 121b:
123a, 123b: a second inlet 2, 124: a second microchannel portion
125: 2nd to 3rd inlet 128: 2nd outlet
130: third layer 133a, 133b: third-
124: third microchannel portion 135: via hole
136: chamber part 138: third outlet

Claims (13)

A first layer 110, a second layer 120, and a third layer 130 stacked on top and bottom,
The first layer is formed with first-1-1 inlet ports (111a, 111b) and first and second inlet ports (113a, 113b)
The second layer is provided with second-1 injection ports 121a and 121b, a second microchannel 124, and a second-third injection port 125 that sequentially communicate with the first-first injection ports 111a and 111b, Is formed,
The third layer is provided with a third microchannel portion 134 and a third microchannel portion 133 which sequentially communicate with the first and second injection ports 113a and 113b of the first layer 110. [ (136) is formed,
Wherein the second microchannel portion and the third microchannel portion are repeatedly joined together with the adjacent channel while one channel is branched. The method according to claim 1,
The second layer is provided with a second 2 inlet 123a communicating with the first 1-2 inlet 113a of the first layer 110 and the 3-1 inlet 133a of the third layer 133b, And 123b are further formed on the surface of the microfluidic channel. 3. The method of claim 2,
The 2-1 inlet (121a, 121b) and the second microchannel (124) are formed on the surface of the second layer,
The micro-algae screening apparatus using the multi-layer microfluidic channel according to claim 1, wherein the second to third injection ports (125) are formed through the second layer. The method of claim 3,
And the second inlet (125) of the second layer is formed to communicate with the chamber (135) of the third layer. The method according to claim 1,
Wherein the chamber part (136) comprises a groove (136a) formed on the surface and a plurality of catching parts (136b) arranged at a predetermined distance in the longitudinal direction of the groove. Screening device. 6. The method of claim 5,
Wherein the capturing unit (136b) is C-shaped. 3. The method of claim 2,
The micro-algae screening apparatus using the multi-layer microfluidic channel according to claim 1, wherein the material injected into the first and second inlets (113a, 113b) of the first layer (110) is microalgae or nutrients. The method of claim 3,
And an outlet through the first layer, the second layer and the third layer is provided for discharging the substance inside the chamber portion. The method according to claim 1,
Further comprising temperature adjusting means for adjusting a temperature of the chamber below the chamber portion of the third layer. A method for screening microalgae using a microalgae screening apparatus comprising a first layer (110), a second layer (120), and a third layer (130) sequentially stacked in a vertical direction,
A first step of injecting microalgae into the first layer;
A second step of passing the microalgae through the second layer and being accommodated in a plurality of chamber parts of the third layer;
A third step of injecting a first nutrient into the first layer;
A fourth step of forming a concentration gradient of the first nutrient through a channel formed on the surface of the second layer, and then injecting the concentration gradient into the chamber part of the third layer;
A fifth step of injecting a second nutrient into the first layer; And
And a sixth step of forming a concentration gradient of the second nutrient through the channel formed on the surface of the third layer and then injecting the concentration gradient into the chamber part of the third layer. 11. The method of claim 10,
Wherein the concentration gradient of the first nutrient is obtained by repeating the process of merging one channel and the neighboring channel while branching. 11. The method of claim 10,
Wherein the concentration gradient of the second nutrient is obtained by repeating the process of merging the channels with neighboring channels while one channel is branched. 11. The method of claim 10,
And controlling the temperature by heating the lower part of the chamber part.

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