CN212128203U - Culture device suitable for research of interaction between cultures through gas communication - Google Patents
Culture device suitable for research of interaction between cultures through gas communication Download PDFInfo
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- CN212128203U CN212128203U CN202020006872.XU CN202020006872U CN212128203U CN 212128203 U CN212128203 U CN 212128203U CN 202020006872 U CN202020006872 U CN 202020006872U CN 212128203 U CN212128203 U CN 212128203U
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Abstract
The utility model relates to the field of biotechnology, a study two kinds of cultures and pass through gaseous interchange culture apparatus of interaction is related to. The culture device comprises a culture bottle I, a culture bottle II and a gas peristaltic pump, wherein a sampling hole is formed in the middle of each culture bottle cap, a gas inlet pipe hole, a gas outlet pipe hole and an oxygen monitor probe inserting hole are formed in the periphery of each sampling hole, and a gas outlet pipe on the culture bottle I is connected with a gas inlet pipe on the culture bottle II through the gas peristaltic pump and a guide pipe; an air inlet pipe I on the culture bottle I is connected with an air outlet pipe II of the culture bottle II through a guide pipe; the pipe orifices of the air inlet pipe and the air outlet pipe are respectively connected with a three-way valve with a screw plug, one end of the three-way valve at the air outlet pipe is connected with a one-way exhaust valve, and the air inlet pipe is provided with threads and is connected with the screw orifice of the air inlet pipe hole. The utility model discloses simple structure, low in manufacturing cost, convenient to use has solved the technical problem that two kinds of cultures exist through gaseous interchange.
Description
Technical Field
The utility model relates to a biotechnology field relates to a study two kinds of cultures and passes through gaseous culture apparatus that exchanges interaction, is particularly useful for studying the culture apparatus that predatory nematode fungi and nematode pass through gaseous signal molecule interact.
Background
Cell and microbial culture techniques are widely used in the fields of medical and biological research. A petri dish is a laboratory vessel used for microbial or cell culture. At present, a typical culture dish consists of a flat disc-shaped bottom and a lid, typically made of glass or plastic. The culture dish is basically made of two types, mainly plastic and glass, and the glass can be used for plant materials, microorganism culture and adherent culture of animal cells. The plastic may be of polyethylene material, disposable and reusable, suitable for laboratory inoculation, streaking, bacterial isolation and may be used for the cultivation of plant material. The culture dish is divided into an independent culture dish and a multi-hole culture dish according to the number of culture holes. Independent culture dishes are generally used by placing cultures on the bottom of the dish, and this conventional method is to culture only one culture. A multi-well culture dish having a plurality of wells for culturing different cultures, but each well is open to each other and is in communication with the outside air and has a relatively small volume. Also use after improving independent culture dish among the prior art, be about to the culture dish inversion, cultivate solid culture at the bottom of the dish, at dish lid culture liquid culture. After the improvement, the two cultures (liquid-solid; solid-solid) can be cultured simultaneously, and the interaction result of the two cultures under the condition of not contacting can also be observed.
Therefore, the culture dishes currently used are independent, do not allow experimental studies of communication between cultures of cells or microorganisms by means of released gases, or have a plurality of wells, each of which is open to each other and is in communication with the outside atmosphere, and are not available for carrying out this type of experiment. Even if the independent culture dish is modified for experimental studies of communication between cultures of cells or microorganisms by releasing gas, the following technical problems still exist:
1. experimental results failed to determine if cells of one culture moved with the gas into another culture;
2. in the cultivation of fungi that produce spores, since they can move in the air, there may also be contamination-related conditions between other bacterial cultures, and it is uncertain whether the changes produced are produced by gas molecules.
The gas generated by the culture cannot be collected, and the interacting vector molecules cannot be identified;
3. the culture dish has limited space, can grow less culture and is not suitable for large-scale culture needing separation and identification of secondary metabolites of the culture.
Disclosure of Invention
To solve the above technical problems, the present invention aims to provide a culture apparatus suitable for studying interaction between cultures through gas communication. The technical scheme is as follows:
a culture device suitable for researching interaction between cultures through gas communication comprises a culture bottle I (1), a culture bottle II (15) and a gas peristaltic pump (2), wherein a sampling hole (4) is formed in the middle of each culture bottle cap, a gas inlet pipe hole (10), a gas outlet pipe hole (8) and an oxygen monitor probe inserting hole (6) are further formed in the periphery of the sampling hole (4), and a gas outlet pipe I on the culture bottle I (1) is connected with a gas inlet pipe II on the culture bottle II (15) through the gas peristaltic pump (2) and a guide pipe; an air inlet pipe I on the culture bottle I (1) is connected with an air outlet pipe II on the culture bottle II (15) through a guide pipe; the mouth of each of the air inlet pipe and the air outlet pipe is respectively connected with a three-way valve (7) with a screw plug, one end of the three-way valve (7) at the air outlet pipe of each culture bottle is connected with a one-way exhaust valve (11), and the oxygen monitor (3) is connected with the culture bottle through a probe inserting hole (6) of the oxygen monitor.
Furthermore, the pipe orifice of the air inlet pipe and the pipe orifice of the air outlet pipe are respectively provided with a microporous filter membrane (14) for filtering microorganisms or cells existing in the air and ensuring the air to pass through.
Further, the gas peristaltic pump belt charges the battery.
Furthermore, a nut with the same diameter as the nut of the probe insertion hole (6) of the oxygen monitor is arranged at the connecting part of the oxygen monitor (3) and the culture bottle.
Furthermore, the air inlet pipe hole (10), the air outlet pipe hole (8) and the oxygen monitor probe insertion hole (6) are uniformly and symmetrically arranged around the culture bottle cap.
Furthermore, the sampling hole (4) and the oxygen monitor probe insertion hole (6) are sealed by screw caps, and the mouth of the culture bottle is connected with the screw cap of the culture bottle.
Furthermore, a pressure detection device (12) is arranged on the conduit beside the gas peristaltic pump (2).
Furthermore, the culture bottle I (1), the culture bottle II (15) and the gas peristaltic pump (2) are placed in a disc (5) provided with a clamping groove with the size suitable for the culture bottle I (1), the culture bottle II (15) and the gas peristaltic pump.
Furthermore, the culture bottle is made of transparent plastic materials which can be sterilized at high temperature and high pressure, and the culture bottle can be conveniently sterilized at high temperature and high pressure.
During operation, add required culture medium in cultivateing I (1) and blake bottle II (15), cover the blake bottle lid, adjust the height of intake pipe, add sample hole (4), the nut is twisted on to oxygen detector probe (6) inserted hole, and the screw plug is twisted on to intake pipe and outlet duct mouth of pipe, inspects the leakproofness of each part of the device. Meanwhile, the screw plug is not required to be screwed too hard, so that the device can exchange air with the outside during high-temperature and high-pressure sterilization, and the explosion caused by too high pressure in the device is avoided. Sterilizing at high temperature and high pressure. The oxygen monitor was irradiated for sterilization alone. And (4) after the culture bottle and the culture medium are cooled, taking down the screw cap of the probe insertion hole of the oxygen monitor in the sterile environment, inserting the probe of the oxygen monitor in the hole, and screwing down the screw cap. And opening the screw cap of the sampling hole, accessing the to-be-cultured substance from the sampling hole, and screwing the screw cap.
The air inlet pipe is set to be adjustable in height, so that the air inlet pipe is close to the surface of the culture solution as much as possible to promote the communication between the gas and the culture solution, according to the experiment requirement, a little more culture medium such as 200ml is needed sometimes, and a little less culture medium such as 50ml is needed sometimes, so that the heights of the culture solution surfaces are different, and therefore the height of the air inlet pipe is adjusted according to the height of the culture solution surface from the bottom of the culture bottle during autoclaving, and the air inlet pipe mouth is enabled to be close to the surface of the culture solution as much as possible.
According to the oxygen consumption condition of the culture, an oxygen concentration monitor is checked at regular time, and the culture with high oxygen consumption needs to be supplemented with oxygen regularly, and the operation is as follows: the three-way valve (7) connected with the air inlet pipe is rotated to a point connected with oxygen, the three-way valve (7) on one side of the air outlet pipe is rotated to one side of the one-way vent valve (11), the one-way vent valve (11) is made to be the same as the device, and the ventilation and the exhaust are stopped when the oxygen concentration is increased back to the normal value range.
When a culture passes a gas interaction experiment, connecting the device, and connecting a gas outlet pipe I on a culture bottle I (1) with a gas inlet pipe II on a culture bottle II (15) (1) through a gas peristaltic pump (2) and a conduit; an air inlet pipe I on the culture bottle I (1) is connected with an air outlet pipe II on the culture bottle II (15) through a guide pipe; all three-way valves are screwed to the state that the culture bottles are connected with the conduits. The gas peristaltic pump (2) is started regularly to promote the gas communication between the two culture bottles. The gas peristaltic pump (2) is provided with a rechargeable battery (13), and the pump can be directly placed in the disc, so that the situation that the power supply is required to be connected and the incubator cannot be closed is avoided. If gas is required to be collected for component analysis, a syringe is connected to the mouth of the one-way vent valve (11) of the culture bottle II (15) (1), the three-way valve (7) is turned to the position where the one-way vent valve (11) is communicated with the culture bottle II (15), the catheter on the air outlet pipe II on the culture bottle II (15) is taken down, the catheter is immersed into liquid (such as ultrapure water or PBS buffer solution), the gas peristaltic pump (2) is started, and the syringe is pulled outwards to draw gas outwards.
In the device, a culture of signal gas molecules which can be possibly generated is placed in a culture bottle I (1) for culture, when the gas generated by culture is accumulated to a certain concentration, a gas outlet pipe I of the culture bottle I (1) and a gas inlet pipe II of a culture bottle II (15) are connected with a gas peristaltic pump (2) through a gas guide pipe, the gas in the culture bottle I (1) is pumped out by the gas peristaltic pump (2) and is transmitted into the culture bottle II (15), the original existing gas in the culture bottle II (15) is introduced, the gas pressure is increased after new gas is introduced, in order to maintain normal gas pressure, the gas needs to be discharged outwards, the gas outlet pipe II of the culture bottle II (15) is connected with the gas inlet pipe I of the culture bottle I (1) through the gas guide pipe, the gas in the culture bottle II (15) can be discharged into the culture bottle I (1), and the gas flows between the culture bottle I (1) and the culture bottle II (15) continuously through the repeated times, the purpose of uniformly distributing the gas generated by the culture in the air of the culture bottle II (15) can be achieved, and the arrangement and the operation can increase the probability of the gas generated by the culture to contact with the culture in the culture bottle II (15), thereby being beneficial to the interaction between the experimental operation and the culture.
Compared with the prior art, the invention has the following beneficial effects:
1. according to the invention, two cultures are separately placed in the culture bottle, and gas communication is promoted by the gas peristaltic pump, so that the problem that whether cells of one culture move to the other culture along with gas cannot be determined is effectively solved. The gas peristaltic pump is used for promoting the gas communication between the two culture bottles, so that the problem of whether the two cultures can be separated or not can be solved.
2. When the fungi capable of producing spores are cultured, because the spores can move in the air, the condition of related pollution can exist between other bacterial cultures, for example, nematodes can climb, the gas peristaltic pump promotes gas communication, so that the two cultures are separated, the problem that the change generated by an experiment cannot be determined is not generated through gas molecules is solved, and microporous filter membranes are arranged at a gas inlet pipe orifice and a gas outlet pipe orifice and used for filtering microorganisms or cells existing in the gas, so that the gas is ensured to pass through, and the interference to the experiment is reduced.
3. The device changes the culture dish into the triangular flask, and the space grow, but the mycelium of cultivateing is many, can carry out liquid culture, collects mycelium and culture solution after the experiment finishes, can solve because by the space restriction, can't develop the problem of the secondary metabolite separation appraisal experiment of fungus.
4. The problem of unable collection culture gas, unable authentication medium molecule is solved.
5. The utility model has simple structure, low manufacturing cost, convenient use, low power consumption and low noise of the gas peristaltic pump; the device is completely closed, and the pollution probability is low during working; the occupied space is small, the culture space is saved, and the experiment cost is reduced.
Drawings and description of the drawings
FIG. 1 is a schematic diagram of the apparatus;
FIG. 2 is a schematic diagram of a partial structure of the apparatus;
the system comprises a culture bottle I, a culture bottle II, a gas peristaltic pump 2, an oxygen monitor 3, a sample adding hole 4, a disc 5, an oxygen monitor probe inserting hole 6, a three-way valve 7, an air outlet pipe hole 8, an air inlet pipe hole 10, a one-way exhaust valve 11, a pressure detection device 12, a rechargeable battery 13 and a microporous filter membrane 14.
Detailed Description
A culture device suitable for researching interaction between cultures through gas communication comprises a culture bottle I1, a culture bottle II15 and a gas peristaltic pump, wherein a sampling hole 4 is formed in the middle of each culture bottle cap, a gas inlet pipe hole 10, a gas outlet pipe hole 8 and an oxygen monitor probe inserting hole 6 are further formed in the periphery of the sampling hole 4, and a gas outlet pipe I on a culture bottle I1 is connected with a gas inlet pipe II on a culture bottle II15 through a gas peristaltic pump 2 and a guide pipe; the air inlet pipe I on the culture bottle I1 is connected with the air outlet pipe II on the culture bottle II15 through a guide pipe; the mouth of each of the air inlet pipe and the air outlet pipe is respectively connected with a three-way valve 7 with a screw plug, one end of the three-way valve 7 at the air outlet pipe of each culture bottle is connected with a one-way exhaust valve 11, and the oxygen monitor 3 is connected with the culture bottle through a probe inserting hole 6 of the oxygen monitor. The mouth of the air inlet pipe and the mouth of the air outlet pipe are respectively provided with a microporous filter membrane 14. The gas peristaltic pump 2 is provided with a rechargeable battery 13. The part of the oxygen monitor 3 where the probe is connected with the culture bottle is provided with a screw cap with the same diameter as the screw cap of the oxygen monitor probe insertion hole 6. The air inlet pipe hole 10, the air outlet pipe hole 8 and the oxygen monitor probe insertion hole 6 are uniformly and symmetrically arranged around the culture bottle cap. The sample adding hole 4 and the probe inserting hole 6 are sealed by nuts. The conduit beside the gas peristaltic pump 2 is also provided with a pressure detection device 12. The mouth of the culture bottle 1 is connected with the screw mouth of the culture bottle cap. The culture bottle I1, the culture bottle II15 and the gas peristaltic pump 2 are placed in a disc 5 provided with a clamping groove with the size suitable for the culture bottle I1 and the culture bottle II 15. The culture bottle is made of transparent plastic materials which can be sterilized at high temperature and high pressure.
During operation, add required culture medium in cultivateing I1 and blake bottle II15, with the blake bottle lid, adjust the height of intake pipe, add the sample hole 4, the nut is twisted on to oxygen detector probe patchhole 6, and the screw plug is twisted on to intake pipe and outlet duct, inspects the leakproofness of each part of the device. Meanwhile, the screw plug is not required to be screwed too hard, so that the device can exchange air with the outside during high-temperature and high-pressure sterilization, and the explosion caused by too high pressure in the device is avoided. Sterilizing at high temperature and high pressure. The oxygen monitor was irradiated for sterilization alone. And (4) after the culture bottle and the culture medium are cooled, taking down the screw cap of the probe insertion hole of the oxygen monitor in the sterile environment, inserting the probe of the oxygen monitor in the hole, and screwing down the screw cap. And opening the screw cap of the sampling hole, accessing the to-be-cultured substance from the sampling hole, and screwing the screw cap.
The air inlet pipe is set to be adjustable in height, so that the air inlet pipe is close to the culture surface as much as possible to promote the communication of gas and culture solution, according to the experiment requirement, a little more culture medium such as 200ml is needed sometimes, and a little less culture medium such as 50ml is needed sometimes, so that the heights of the culture liquid surfaces are different, and therefore the height of the air inlet pipe is adjusted according to the height of the culture medium liquid surface away from the bottom of the culture bottle during autoclaving, and the air inlet pipe mouth is enabled to be close to the culture solution surface as much as possible.
According to the oxygen consumption condition of the culture, an oxygen concentration monitor is checked at regular time, and the culture with high oxygen consumption needs to be supplemented with oxygen regularly, and the operation is as follows: the three-way valve 7 connected with the air inlet pipe is rotated to one side connected with oxygen, the three-way valve 7 at one side of the air outlet pipe is rotated to one side of the one-way vent valve 11, the one-way vent valve 11 is communicated with the device, and the ventilation and the exhaust are stopped when the oxygen concentration is increased back to the normal value range.
When the culture passes through a gas interaction experiment, the device is connected, and a gas outlet pipe I on a culture bottle I1 is connected with a gas inlet pipe II on a culture bottle II15 through a gas peristaltic pump 2 and a guide pipe; the air inlet pipe I on the culture bottle I1 is connected with the air outlet pipe II on the culture bottle II15 through a guide pipe; all three-way valves are screwed to the state that the culture bottles are connected with the conduits. The gas peristaltic pump 2 is started at regular time to promote gas communication between the two culture bottles. The gas peristaltic pump 2 is provided with a rechargeable battery 13, and the pump can be directly placed in a disc, so that the situation that a power supply is required to be connected and an incubator cannot be closed is avoided. If gas is required to be collected for component analysis, a syringe is connected to the port of the one-way vent valve 11 of the culture bottle II15, the three-way valve 7 is turned to the position where the one-way vent valve 11 is communicated with the culture bottle II15, the catheter on the outlet pipe II on the culture bottle II15 is taken down, the catheter is immersed into liquid (such as ultrapure water or PBS buffer), the gas peristaltic pump 2 is started, and the syringe is pulled outwards to draw gas outwards.
In the device, a culture which can generate gas signal molecules is placed in a culture bottle I1 for culture, when the gas generated by culture is accumulated to a certain concentration, an air outlet pipe I of the culture bottle I1 and an air inlet pipe II of a culture bottle II15 are connected with each other through an air guide pipe and a gas peristaltic pump 2, the gas in the culture bottle I1 is pumped by the gas peristaltic pump 2 and is transmitted into the culture bottle II15, the gas originally existing in the culture bottle II15 is present, the air pressure is increased after new gas is introduced, in order to maintain normal air pressure, the gas needs to be discharged outwards, the air outlet pipe II of the culture bottle II15 is connected with the air inlet pipe I of the culture bottle I1 through the air guide pipe, the gas in the culture bottle II15 can be discharged into the culture bottle I1, the gas can continuously flow between the culture bottle I1 and the culture bottle II15 for a plurality of times, the aim of uniformly distributing the gas generated by culture in the culture bottle II15 can be achieved, and the arrangement and the operation can increase the probability of the contact of the gas generated by culture bottle II15, is beneficial to the interaction between experimental operation and culture.
Compared with the prior art, the invention has the following beneficial effects:
1. according to the invention, two cultures are separately placed in the culture bottle, and gas communication is promoted by the gas peristaltic pump, so that the problem that whether cells of one culture move to the other culture along with gas cannot be determined is effectively solved. The gas peristaltic pump is used for promoting the gas communication between the two culture bottles, so that the problem of whether the two cultures can be separated or not can be solved.
2. When the fungi capable of producing spores are cultured, because the spores can move in the air, the condition of related pollution can exist between other bacterial cultures, for example, nematodes can climb, the gas peristaltic pump promotes gas communication, so that the two cultures are separated, the problem that the change generated by an experiment cannot be determined is not generated through gas molecules is solved, and microporous filter membranes are arranged at a gas inlet pipe orifice and a gas outlet pipe orifice and used for filtering microorganisms or cells existing in the gas, so that the gas is ensured to pass through, and the interference to the experiment is reduced.
3. The device changes the culture dish into the triangular flask, and the space grow, but the mycelium of cultivateing is many, can carry out liquid culture, collects mycelium and culture solution after the experiment finishes, can solve because by the space restriction, can't develop the problem of the secondary metabolite separation appraisal experiment of fungus.
4. The problem of unable collection culture gas, unable authentication medium molecule is solved.
5. The utility model has simple structure, low manufacturing cost, convenient use, low power consumption and low noise of the gas peristaltic pump; the device is completely closed, and the pollution probability is low during working; the occupied space is small, the culture space is saved, and the experiment cost is reduced.
The device is not limited to the research on the interaction between the nematodes and the nematode-trapping fungi, and can also be used for the research on the interaction between the nematodes and bacteria, the interaction between the bacteria and fungi, the interaction between the fungi and fungi, and the interaction between the bacteria and animal cells by taking volatile gas as a medium.
Claims (9)
1. A culture device adapted for studying interactions between cultures through gas communication, comprising: the culture device comprises a culture bottle I (1), a culture bottle II (15) and a gas peristaltic pump (2), wherein a sampling hole (4) is formed in the middle of each culture bottle cap, a gas inlet pipe hole (10), a gas outlet pipe hole (8) and an oxygen monitor probe inserting hole (6) are further formed in the periphery of the sampling hole (4), and the gas outlet pipe I on the culture bottle I (1) is connected with the gas inlet pipe II on the culture bottle II (15) through the gas peristaltic pump (2) and a guide pipe; an air inlet pipe I on the culture bottle I (1) is connected with an air outlet pipe II on the culture bottle II (15) through a guide pipe; the mouth of each of the air inlet pipe and the air outlet pipe is respectively connected with a three-way valve (7) with a screw plug, one end of the three-way valve (7) at the air outlet pipe of each culture bottle is connected with a one-way exhaust valve (11), and the oxygen monitor (3) is connected with the culture bottle through a probe inserting hole (6) of the oxygen monitor.
2. Culture device suitable for studying the interaction between cultures by gas communication according to claim 1, characterized in that: the mouth of the air inlet pipe and the mouth of the air outlet pipe are respectively provided with a microporous filter membrane (14).
3. Culture device suitable for studying the interaction between cultures by gas communication according to claim 1, characterized in that: the gas peristaltic pump (2) is provided with a rechargeable battery (13).
4. Culture device suitable for studying the interaction between cultures by gas communication according to claim 1, characterized in that: the part of the oxygen monitor (3) connected with the culture bottle is provided with a screw cap with the same diameter as the screw cap of the probe insertion hole (6) of the oxygen monitor.
5. Culture device suitable for studying the interaction between cultures by gas communication according to claim 1, characterized in that: the air inlet pipe hole (10), the air outlet pipe hole (8) and the oxygen monitor probe insertion hole (6) are uniformly and symmetrically arranged around the culture bottle cap.
6. Culture device suitable for studying the interaction between cultures by gas communication according to claim 1, characterized in that: the sampling hole (4) and the oxygen monitor probe insertion hole (6) are sealed by screw caps, and the mouth of the culture bottle is connected with the screw mouth of the culture bottle cap.
7. Culture device suitable for studying the interaction between cultures by gas communication according to claim 1, characterized in that: the conduit beside the gas peristaltic pump (2) is also provided with a pressure detection device (12).
8. Culture device suitable for studying the interaction between cultures by gas communication according to claim 1, characterized in that: the culture bottle I (1), the culture bottle II (15) and the gas peristaltic pump (2) are placed in a disc (5) provided with a clamping groove with the size suitable for the culture bottle I (1) and the culture bottle II (15).
9. Culture device suitable for studying the interaction between cultures by gas communication according to claim 1, characterized in that: the culture bottle is made of transparent plastic materials which can be sterilized at high temperature and high pressure.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020006872.XU CN212128203U (en) | 2020-01-03 | 2020-01-03 | Culture device suitable for research of interaction between cultures through gas communication |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020006872.XU CN212128203U (en) | 2020-01-03 | 2020-01-03 | Culture device suitable for research of interaction between cultures through gas communication |
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| Publication Number | Publication Date |
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| CN212128203U true CN212128203U (en) | 2020-12-11 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202020006872.XU Expired - Fee Related CN212128203U (en) | 2020-01-03 | 2020-01-03 | Culture device suitable for research of interaction between cultures through gas communication |
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| Country | Link |
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| CN (1) | CN212128203U (en) |
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2020
- 2020-01-03 CN CN202020006872.XU patent/CN212128203U/en not_active Expired - Fee Related
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Granted publication date: 20201211 Termination date: 20220103 |