KR101155137B1 - Micro plate for bioreactor - Google Patents

Abstract

The present invention relates to a microplate for a bioreactor, and more particularly, for a bioreactor capable of preventing contamination of a culture medium cultured in a well provided in a microplate with air from a culture medium or a chamber of an adjacent well. Relates to a microplate.

The microplate of the present invention includes a frame that can be mounted in a chamber, a plurality of wells provided in the frame and a culture space for culturing the microorganism therein, and an upper portion of which is open, and an upper portion of at least one of the wells. Is coupled to aeration between the culture space and the outside of the well, but the foreign matter is provided with a stopper for filtering.

Bioreactor, Reactor, Culture, Microplate

Description

Micro plate for bioreactor

The present invention relates to a microplate for a bioreactor, and more particularly, for a bioreactor capable of preventing contamination of a culture medium cultured in a well provided in a microplate with air from a culture medium or a chamber of an adjacent well. Relates to a microplate.

Bioprocess technology has emerged as an economically important technology as the production of fine chemical products and the production of therapeutic agents such as enzymes and recombinant proteins have increased. Bioprocess technology, which has shown remarkable growth in the 21st century, is expected to have high expected value in the future industrial society.

As knowledge of microorganisms accumulate, efforts are being made to convert and manipulate them in a way that is useful for humankind. As a result, researches on bioreactors are being actively conducted to raise specific organisms in a controlled environment. . Bioreactors are reactors that allow organisms to be raised in a controlled environment to produce specific substances or cells or to perform specific reactions.

In particular, due to the various capabilities of microorganisms, the use of waste water treatment, carbon dioxide immobilization, fuel material production, pharmaceuticals, cosmetics, etc. has been expanded, and the demand is increasing. However, development of economically and technically efficient incubators is urgently needed to produce large quantities of biomass and useful products.

Recently, various high value products derived from microorganisms have been developed, and various culture apparatuses for high concentration culture thereof have been developed. In warm climates and low land costs, such as in the equator, outdoor cultures in the form of ponds or raceways that circulate the medium to paddl wheels are used. While the operating cost is low, the high concentration is difficult and the possibility of contamination by other microorganisms increases the cost of separation and recovery of the target product.

In order to overcome this, various types of bioreactors suitable for high concentration and mass culture have been developed. However, all existing bioreactors are used for mass production of biological products in large quantities, and due to the size of the fermenter, a lot of time and labor are required, and expensive substrates consume many disadvantages.

Therefore, there is a growing demand for a multi-channel bioreactor capable of miniaturizing and speeding up the process to have a simple structure for optimizing a biological production process and to facilitate manipulation and to search for a new photosynthetic strain.

Korean Patent No. 10-0860958 discloses a multi-channel small bioreactor attached to an optical sensor film. The bioreactor has an effect of obtaining or monitoring a plurality of photodetection results in one bioreactor using a microplate in which a plurality of wells are formed.

However, since the microplates are formed with a plurality of wells adjacent to each other, there is a problem that the culture medium cultured inside each well may be contaminated by different culture fluids or the culture fluid may be contaminated by air inside the chamber in which the microplate is mounted. have.

SUMMARY OF THE INVENTION The present invention has been made to improve the above problems, and provides a microplate for a bioreactor capable of preventing the culture medium cultured in the well from being contaminated from the culture medium of the adjacent wells or the air inside the chamber. There is a purpose.

Microplate for a bioreactor of the present invention for achieving the above object is a microplate for culturing microorganisms is installed in the chamber of the bioreactor, the frame which can be mounted to the chamber; A plurality of wells provided in the frame and having a culture space for culturing the microorganism therein and having an open top; And a stopper coupled to an upper portion of at least one of the wells to allow the culture space and the outside of the wells to be vented with foreign substances filtered.

The stopper includes an insertion part which is hollow inside and is inserted into the well; It is characterized in that it comprises a porous ventilation pad to be installed in, and a sealing member which is installed on the outer peripheral surface of the insert portion in close contact with the inner peripheral surface of the well.

And a light transmitting unit installed in the stopper so that light irradiated from the light source installed in the chamber can be transferred to the culture space of the well.

The light transmitting unit is inserted into a plurality of insertion holes formed through the head and the insertion unit, respectively, and has one end exposed to the outside of the head and the other end provided with optical fibers exposed below the insertion unit.

As described above, according to the present invention, it is possible to prevent contamination by adjacent wells by culturing a stopper when culturing microorganisms in a well of a microplate. In addition, the stopper is made of a porous structure that can be vented with the outside air and bet to allow the culture of aerobic microorganisms.

The light generated by the light source may be delivered to the culture space of the well by the phototransmitter formed on the stopper, thereby cultivating photosynthetic microorganisms.

Hereinafter, a microplate for a bioreactor according to a preferred embodiment of the present invention with reference to the accompanying drawings will be described in detail.

1 is a perspective view illustrating a state in which a microplate is mounted in a chamber according to an embodiment of the present invention, FIG. 2 is an exploded perspective view illustrating the microplate of FIG. 1, and FIG. 3 is a main portion of the microplate of FIG. 1. Here is an excerpt.

1 to 3, the microplate according to an embodiment of the present invention is installed in the chamber 5 of the bioreactor 1 and used for culturing microorganisms. As an example of the bioreactor 1, a chamber 5 is provided at an upper side, and a door 7 capable of opening and closing the chamber 5 is provided at an upper portion of the chamber 5. The bottom of the chamber 5 is mounted with the microplate 10 of the present invention.

In the micro plate 10 of the present invention, a plurality of wells 25 are formed in one frame 20. Multiple wells 25 are aligned in the frame 20. In the illustrated example, the microplate 10 is formed of 24 wells 25, but various wells 25 may be formed in addition to 4 to 1536.

The well 25 has a form in which a culture space 27 for culturing microorganisms is formed and an upper portion thereof is opened. The well 25 may be any shape such as a square column, a cylinder, a rhombus column, a test tube, and the like as long as the culture space in which the culture medium is filled is formed.

The microplate 10 of the present invention has a stopper 30 coupled to the top of the well 25. The stopper 30 may be provided in the same number as the number of the wells 25, and may be coupled to each well 25, or the stopper may be coupled to any one well or two or more wells. The stopper 30 allows the culture space of any well 25 to be independent from the adjacent wells. This is to prevent contamination by the culture of adjacent wells. In addition, the stopper 30 serves as a filtering to prevent foreign matter from entering the air into the culture space of the well 25.

The stopper 30 is installed in the insertion part 31, the head 35 formed on the insertion part 31, the ventilation pad 50 installed on the head 35, and the insertion part 31, for example. It is made of a sealing member 40. The insertion part 31 and the head 35 may be injection molded integrally using a Teflon material, but may have a structure in which the insert part 31 and the head 35 are coupled to each other.

Insertion portion 31 is formed in a cylindrical shape hollow inside. The sealing member 40 is mounted on the outer circumferential surface of the insert 31. As the sealing member 40, a conventional rubber ring, pad, or the like may be used. In the illustrated example, a rubber ring is applied to the sealing member 40. In order to install the sealing member 40 in the inserting portion, the fitting groove 33 is formed on the outer circumferential surface of the inserting portion 31. Insert portion 31 is formed in a cylindrical shape corresponding to the shape of the culture space of the well 25, in addition, if the culture space 27 is formed in a shape other than the cylindrical insert 31 may also be formed in a different shape Of course. For example, when the culture space of the well 25 is formed in a square, the insertion portion 31 is also formed in a square. The outer diameter of the insert 31 is formed slightly smaller than the inner diameter of the well 25. The gap generated between the inner circumferential surface of the well 25 and the outer circumferential surface of the insert 31 is sealed by the sealing member 40. If the inserting portion 31 is formed of an elastic material, it is possible to omit the installation of the sealing member 40 by forming a frame projecting outward on the outer circumferential surface of the inserting portion 31.

The head 35 is formed on the upper portion of the insertion portion 31 and extends upward by a predetermined height. The head 35 is formed with an outer diameter larger than the outer diameter of the insert 31. Therefore, the head 35 is formed to have an enlarged cross-sectional area on the upper portion of the insertion portion 31. When the stopper 30 is coupled to the top of the well 25, the top of the well 25 is caught by the head 35. Inside the head 35, a mounting hole 37 to which the ventilation pad 50 is mounted is formed. The mounting hole 37 is formed through the top and bottom of the head 35 so as to be connected to the space inside the insertion portion 31. Preferably, the diameter of the mounting hole 37 of the head is larger than the diameter of the circular space formed inside the insertion portion 31. Therefore, since the jaw 39 is formed inside the stopper 30, the ventilation pad 50 is easily mounted. The mounting hole 37 also has the same shape as the ventilation pad 50.

Ventilation pad 50 is made of porous pores formed with countless pores to vent between the culture space 27 and the outside of the well, but the foreign matter can be filtered. The pores formed in the vent pad 50 preferably have a size of 1 탆 or less on average. Ventilation pad 50 may be a porous silicone or foamed resin, sponge or the like.

As described above, the present invention can effectively prevent contamination by adjacent wells by culturing the microorganisms by coupling the stopper 30 to the top of the well 25. In addition, the stopper 30 is made of a porous structure that can be vented to the outside and the bet to allow the culture of aerobic microorganisms.

A stopper according to another example of the invention is shown in FIG. Referring to FIG. 4, a light transmitting unit is installed in the head 35 so that light irradiated from a light source installed in the chamber may be transferred into the culture space of the well. Although a light emitting diode device is most suitable as a light source, at least one selected from a light source for emitting photosynthetically active radiation (PAR) such as a fluorescent lamp, a halogen lamp, an optical fiber, and a neon tube may be used.

The light transmitting unit transfers the light generated by the light source to the well culture space so that the light can be well transmitted to the light source culture solution. Such a phototransmitter allows the photosynthetic microorganism to be cultured in a well culture space. There are many factors that influence the growth of biomass and useful products in cultivation of photosynthetic microorganisms, including the composition of the medium, temperature, pH, light intensity, light quantity, etc. do. Therefore, the supply of light is an important factor to be considered in the cultivation of photosynthetic microorganisms.

In the example shown, the light transmitting portion consists of a plurality of optical fibers 60. The optical fiber 60 is inserted into each of the plurality of insertion holes formed through the head 35 and the insertion portion 31 vertically. One end of the optical fiber 60 is exposed upward of the head 35 and the other end is exposed downward of the insertion part 31. Therefore, the light generated from the light source is incident to one end of the optical fiber 60 and is emitted to the culture space through the other end of the optical fiber. In order to diffuse light emitted from the other end of the optical fiber, a diffusion lens may be installed on the bottom surface of the insertion part 31.

As the optical fiber 60, glass optical fiber or plastic optical fiber can be used. In particular, it is preferable to use a plastic optical fiber having excellent optical properties and processability than glass optical fiber. Plastic optical fiber has a very large ratio of core to cross section. For example, 1mm Out-dia standard fiber dia is 0.98mm, Clad dia is 0.02mm, so the ratio of core to fiber cross-sectional area is 98%. This means that the light transmission efficiency of the plastic optical fiber is very high. In addition, the plastic optical fiber has the advantages of low cost and easy installation due to excellent bending characteristics, and strong external shock. The plastic optical fiber consists of a core made of high-purity acrylic resin (PMMA: Polymethyl methacrylate) and a thin clad layer made of special fluoropolymer (F-PMMA: Fluorine Polymethyl methacrylate). Since the refractive index of the clad is lower than that of the core, light from one end of the optical fiber causes total reflection at the interface between the core and the clad, and is transmitted through the core to the other end.

Unlike in FIG. 4, the optical fiber 60 does not penetrate the head 35 and the insert 31 up and down, but one end of the optical fiber 60 may be installed to be exposed to the side of the head 35. . Therefore, the optical fiber 60 is bent in a translator (a) type is installed on the stopper. This is advantageous when light is incident on the side of the head 35. In this way, the stopper allows the inside and outside air to be ventilated by the ventilation pads, and at the same time, light may be transferred from the outside of the well to the inside by the optical fiber 60.

On the other hand, Figure 5 shows an example in which the light transmission unit is installed in the vent pad 50 of the stopper. Referring to FIG. 5, the optical fiber 60 is installed at each of the plurality of insertion holes 70 formed adjacent to the outer circumference of the vent pad 50. At this time, if the thickness of the ventilation pad 50 is thin or the material itself is not easy to fix the optical fiber 60 in the ventilation pad 50 is preferably provided with a holder 80 that can support the optical fiber. Do. Therefore, the optical fiber 60 is inserted into the holder 80 to fix the optical fiber 60, and then the holder 80 is inserted into the insertion hole 70 to be installed. The holder 80 is composed of a cylindrical body 85 and a locking portion 81 formed on the upper portion of the body 85 and having a diameter larger than the diameter of the body 85. A hole penetrating from the locking portion 81 to the body 85 is formed in the holder 80. The optical fiber 60 is inserted into the hole. In this way, the optical fiber 60 can be easily fixed to the ventilation pad 50 by the holder 80.

In the above, the present invention has been described with reference to one embodiment shown in the drawings, but this is merely exemplary, and it will be understood by those skilled in the art that various modifications and equivalent embodiments are possible. will be.

Therefore, the true scope of protection of the present invention should be defined only by the appended claims.

1 is a perspective view illustrating a state in which a microplate according to an embodiment of the present invention is mounted in a chamber.

2 is an exploded perspective view illustrating the microplate of FIG. 1;

3 is a cross-sectional view of the main portion of the microplate of FIG.

4 is a partially cutaway perspective view showing a stopper applied to a microplate according to another embodiment of the present invention;

5 is an exploded perspective view illustrating a ventilation pad applied to a micro plate according to another embodiment of the present invention.

<Description of the symbols for the main parts of the drawings>

1: bioreactor 5: chamber

10: microplate 20: frame

25: well 27: culture space

30: stopper 31: insert

35: head 40: sealing member

50: aeration pad

Claims (4)

delete delete delete In the microplate for culturing microorganisms installed in the chamber of the bioreactor, A frame that can be mounted to the chamber; A plurality of wells provided in the frame and having a culture space for culturing the microorganism therein and having an open top; And a stopper coupled to an upper portion of at least one of the wells to allow the culture space and the outside of the well to pass through, but filtering foreign matters. The stopper includes an insertion part which is hollow inside and is inserted into the well, a head which is formed on an upper part of the insertion part and has a diameter larger than that of the insertion part and is caught on the top of the well, and a mounting hole penetrating the head up and down. And a porous ventilation pad installed on the sealing member and installed on an outer circumferential surface of the insertion part to be in close contact with the inner circumferential surface of the well. And a light transmitting unit installed in the stopper so that light irradiated from the light source installed in the chamber can be transferred to the culture space of the well. The optical transmission unit has optical fibers whose one end is exposed above the ventilation pad and the other end is exposed below the ventilation pad, and the hole into which the optical fiber is inserted so as to be installed on the ventilation pad to support the optical fibers. Having holders formed therethrough, Each holder includes a cylindrical body inserted into an insertion hole formed by penetrating the ventilation pad up and down, and a locking portion formed on an upper portion of the body and having a diameter larger than the diameter of the body to be caught by the ventilation pad. Microplate for a bioreactor, characterized in that.

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