A sealed microbial culture device and a microbial culture method

Abstract

The application provides a sealed microbial culture device and a microbial culture method. The device comprises a sealed culture box, an environment control assembly, an air inlet assembly, a liquid inlet assembly and a sampling assembly. The sealed culture box is transparent and used for observing and culturing microorganisms. The environment control assembly is used for collecting and adjusting the environmental parameters in the sealed culture box. The air inlet assembly is used for providing gas into the sealed culture box. The liquid inlet assembly is used for providing liquid into the sealed culture box. The sampling assembly is arranged on the outer wall of the sealed culture box and used for extracting or discharging the gas or liquid in the sealed culture box. The application can realize the culture of microorganisms in a specific sealed environment, can adjust the environmental factors, gas and liquid factors in the sealed culture box, and avoids the influence of the gas extraction and liquid extraction process on the culture environment. The application has the characteristics of convenient operation, easy cleaning, good sealing, high use efficiency and continuous experiment.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbial culture, and in particular to an environment factor adjustable closed microbial culture device and a microbial culture method. BACKGROUND

[0002] Microorganisms are a large group of organisms including bacteria, viruses, fungi, and some small protists, microscopic algae, etc. They are tiny in size, but have a great impact on the evolution of the earth's environment, the formation of mineral resources, changes in ecological environment, industrial and agricultural production, human health, etc. Research related to microorganisms has now involved many fields such as petroleum, mineral resources, chemical industry, biology, medicine, food, industry and agriculture, and environmental protection. Microbial culture in specific environments is also considered an important technical means of current scientific research.

[0003] The survival environment of microorganisms includes temperature, pressure, gas, water, light, and many other environmental factors, which directly affect the efficiency of microbial culture. However, these environmental factors may not be the same as the current environment of the earth. In order to correctly understand the survival environment and mechanism of microorganisms, it is necessary to adjust the environmental factors of the closed microbial culture. The existing commercially available microbial incubators are mostly heating type constant temperature incubators, and the adjustable environmental factors are very limited. For example, the Chinese invention patent with the application number 201810487432.8 and the invention name of a constant temperature microbial incubator to avoid cross contamination can be simply adjusted in terms of temperature, light and other environmental parameters, but the parameters related to the survival environment of microorganisms such as atmosphere and water cannot be freely set.

[0004] Microbial culture is generally a continuous experiment within a certain time range, and multiple groups of comparison are needed to master the growth curve of microorganisms under different environmental factors and to select the most favorable environmental factors. The existing closed microbial culture incubator provides a box space for placing microbial culture dishes, but it is not a direct culture space. Therefore, during the continuous opening and sampling of the incubator, the atmosphere and water environment of the culture dish will be disturbed, which is easy to cause cross contamination or loss of control of the culture environment, and thus affect the culture results. In order to avoid cross contamination, experimenters often need to culture multiple microbial samples under the same environmental factor. This makes the process of microbial culture experiment with special environmental factors extremely tedious, and it is also difficult to ensure that the parameters of the parallel samples set under the same environmental factor are strictly consistent. Moreover, the number of microbial culture incubators used in the experiment process will increase exponentially, but the efficiency is very low. SUMMARY

[0005] The present application is used to solve the problems of limited adjustable environmental factors, low use efficiency and not conducive to microbial culture of the existing microbial culture incubator.

[0006] To solve the above technical problems, the first aspect of the present application provides a sealed microbial culture device, comprising: a sealed incubator, an environmental control assembly, an air inlet assembly, a liquid inlet assembly, a sampling assembly;

[0007] The sealed incubator is a transparent structure for observing and culturing microorganisms.

[0008] The environmental control assembly is arranged at the bottom of the sealed incubator for collecting and adjusting the environmental parameters in the sealed incubator.

[0009] The air inlet assembly is arranged on the outer wall of the sealed incubator for providing gas into the sealed incubator.

[0010] The liquid inlet assembly is arranged on the outer wall of the top of the sealed incubator for providing liquid into the sealed incubator.

[0011] The sampling assembly is arranged on the outer wall of the sealed incubator for extracting or discharging gas or liquid in the sealed incubator; wherein the sampling assembly comprises a first valve, a second valve and a sampling bin.

[0012] The first valve is arranged on the outer wall of the sealed incubator; the second valve is provided with a sampling port for connecting a sampling device; and the sampling bin is arranged between the first valve and the second valve for containing the sampled gas or liquid.

[0013] In further embodiments of the present application, the sealed incubator is integrally formed by a transparent plate or sealed by multiple transparent plates, and the bottom of the sealed incubator is heat-conducting.

[0014] In further embodiments of the present application, the environmental control assembly comprises a temperature control unit, a light control unit and a pressure control unit.

[0015] The temperature control unit is used for adjusting and measuring the temperature in the sealed incubator.

[0016] The light control unit is used for adjusting and measuring the light in the sealed incubator.

[0017] The pressure control unit is used for adjusting and measuring the pressure in the sealed incubator.

[0018] In further embodiments of the present application, the sampling assembly comprises a gas sampling assembly and a liquid sampling assembly, wherein the liquid sampling assembly further comprises a third valve arranged between the first valve and the sampling bin or arranged between the sampling bin and the second valve for inputting cleaning liquid to clean the sampling bin.

[0019] In further embodiments, the closed microbial culture device further comprises a microwave oscillation assembly arranged at the bottom of the closed culture box, for mixing the liquid in the closed culture box by microwave oscillation.

[0020] In further embodiments, the closed microbial culture device further comprises a support cabinet arranged at the lower part of the closed culture box, for supporting the closed culture box and accommodating the environmental control assembly.

[0021] In further embodiments, the closed microbial culture device further comprises a display and control assembly arranged on the support cabinet, connected to the environmental control assembly, for displaying the environmental parameters in the closed culture box and controlling the operation of the environmental control assembly.

[0022] In a second aspect, a microbial culture method is provided, which is applicable to the closed microbial culture device of any of the preceding embodiments, comprising:

[0023] injecting a cleaning liquid into the closed culture box through the liquid injection assembly, and discharging the cleaning liquid in the closed culture box through the sampling assembly after cleaning;

[0024] injecting an experimental liquid into the closed culture box through the liquid injection assembly, and injecting a deoxygenation gas into the closed culture box through the gas injection assembly to perform deoxygenation treatment on the experimental liquid;

[0025] injecting an experimental gas into the closed culture box through the gas injection assembly;

[0026] injecting a microbial culture medium into the closed culture box through the liquid injection assembly;

[0027] setting the environment in the closed culture box through the environmental control assembly;

[0028] adding microorganisms into the closed culture box through the liquid injection assembly to start the culture of microorganisms.

[0029] In further embodiments, after starting the culture of microorganisms, the method further comprises:

[0030] extracting the gas in the closed culture box through the gas extraction assembly to analyze the influence of microbial activity on the gas according to the extracted gas.

[0031] In further embodiments, after starting the culture of microorganisms, the method further comprises:

[0032] extracting the liquid in the closed culture box through the liquid extraction assembly to analyze the influence of microbial activity on the liquid according to the extracted liquid.

[0033] This paper presents a closed-loop microbial culture device and method. Through the configuration of a closed incubator, environmental control components, air intake components, and liquid intake components, it enables the cultivation of microorganisms in a specific closed environment. Environmental factors within the closed incubator, as well as gas and liquid factors, can be adjusted. The sampling components allow for contactless gas and liquid sampling, avoiding any impact on the culture environment during these processes. Furthermore, the air intake, liquid intake, and sampling components enable contactless cleaning of the closed incubator, allowing for repeated experiments. This method is characterized by convenient operation, easy cleaning, good airtightness, high efficiency, and continuous experimental capability, and can be applied in fields such as petroleum, mining, chemical, biological, pharmaceutical, food, industry, agriculture, and environmental protection.

[0034] To make the above and other objects, features and advantages of this document more apparent and understandable, preferred embodiments are described below in detail with reference to the accompanying drawings. Attached Figure Description

[0035] To more clearly illustrate the technical solutions in the embodiments or prior art described herein, the accompanying drawings used in the description of the embodiments or prior art will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this article. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0036] Figure 1 This paper presents a three-dimensional structural schematic diagram of the closed microbial culture device according to the embodiments of this paper;

[0037] Figure 2 This is a front view schematic diagram of the closed microbial culture device according to the embodiments of this article;

[0038] Figure 3 This is a top view schematic diagram of the closed microbial culture device according to the embodiments of this article;

[0039] Figure 4 A side view structural schematic diagram of the closed microbial culture device of the embodiments herein is shown;

[0040] Figure 5 A side view of the gas extraction assembly in the embodiment of this paper is shown.

[0041] Figure 6A and Figure 6B A side view of the liquid collection assembly in the embodiment of this paper is shown.

[0042] Figure 7 A first flowchart of the microbial culture method described in this embodiment is shown;

[0043] Figure 8A second flow chart of the microbial culture method of the embodiments herein is shown.

[0044] List of Symbols:

[0045] 100, closed incubator

[0046] 200, environmental control assembly

[0047] 300, air inlet assembly

[0048] 400, liquid inlet assembly

[0049] 500, sampling assembly

[0050] 510, air sampling assembly

[0051] 520, liquid sampling assembly

[0052] 210, temperature control unit

[0053] 220, light control unit

[0054] 230, pressure control unit

[0055] 511, first valve

[0056] 512, second valve

[0057] 513, air sampling chamber

[0058] 521, first valve

[0059] 522, second valve

[0060] 523, liquid sampling chamber

[0061] 524, third valve

[0062] 600, microwave oscillation assembly

[0063] 700, support cabinet

[0064] 800, display and control assembly

[0065] 900, power switch button DETAILED DESCRIPTION

[0066] The technical solutions in the embodiments herein will be described clearly and completely below with reference to the drawings in the embodiments herein. Obviously, the described embodiments are only part of the embodiments herein, rather than all the embodiments. Based on the embodiments herein, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection herein.

[0067] The existing microorganism incubator is only a box space for placing a microorganism culture dish, and cannot realize the setting of the microorganism culture environment, gas and liquid. In addition, the existing microorganism incubator is taken by opening, which is easy to cause cross contamination or environmental out of control, and then affects the culture effect.

[0068] In order to solve the above technical problems in the prior art, in an embodiment of the present application, a completely closed microorganism culture device is provided, as shown in the figure, comprising: a closed incubator 100, an environment control assembly 200, an air inlet assembly 300, a liquid inlet assembly 400, and a sampling assembly 500. Figure 1

[0069] The closed incubator 100 is a transparent structure for observing and culturing microorganisms.

[0070] The environment control assembly 200 is arranged at the bottom of the closed incubator 100, and is used for collecting and adjusting the environmental parameters in the closed incubator 100.

[0071] The air inlet assembly 300 is arranged on the outer wall of the closed incubator 100, and is used for providing gas into the closed incubator 100.

[0072] The liquid inlet assembly 400 is arranged on the top or side wall of the closed incubator 100, and is used for providing liquid into the closed incubator 100.

[0073] The sampling assembly 500 is arranged on the outer wall of the closed incubator 100, and is used for extracting or discharging the gas or liquid in the closed incubator 100.

[0074] In specific implementation, in order to avoid the mutual interference of gas and liquid sampling, the sampling assembly 500 comprises a gas sampling assembly and a liquid sampling assembly. The gas sampling assembly is arranged on the top outer wall of the closed incubator 100, and is used for extracting or discharging the gas in the closed incubator 100. The liquid sampling assembly is arranged on the bottom outer wall of the closed incubator 100, and is used for extracting or discharging the liquid in the closed incubator 100.

[0075] The closed microorganism culture device provided in the embodiment can realize the culture of microorganisms in a specific closed environment, can adjust the environmental factors, gas and liquid factors in the closed incubator, can realize non-contact gas sampling and liquid sampling through the arrangement of the gas sampling assembly and the liquid sampling assembly, and can avoid the influence of the gas sampling and liquid sampling process on the culture environment. In addition, through the arrangement of the air inlet assembly, the liquid inlet assembly and the liquid sampling assembly, the closed incubator can be cleaned non-contactly, so that repeated experiments can be realized. The present application has the characteristics of convenient operation, easy cleaning, good sealing, high use efficiency and continuous experiment, and can be applied in the fields of petroleum, mineral, chemical industry, biology, medicine, food, industry and agriculture, and environmental protection.​

[0076] In one embodiment of this document, to facilitate observation of the microbial culture process, the sealed incubator 100 is made of a transparent sheet (e.g., plexiglass). Specifically, it can be integrally molded from a transparent sheet or formed by sealing and gluing multiple transparent sheets together; this document does not limit the specific implementation. Furthermore, to improve the heating capacity of the sealed incubator 100, the bottom of the sealed incubator 100 is heat-conducting. To ensure that the sealed incubator 100 is not easily broken, its thickness is, for example, approximately 12 mm.

[0077] In one embodiment of this article, such as Figure 2 , Figure 3 and Figure 4 As shown, the environmental control component 200 includes: a temperature control unit 210, a light control unit 220, and a pressure control unit 230;

[0078] Temperature control unit 210 is used to adjust and measure the temperature inside the sealed incubator 100. Light control unit 220 is used to adjust and measure the light intensity inside the sealed incubator 100. Pressure control unit 230 is used to adjust and measure the pressure inside the sealed incubator 100. The temperature accuracy is 0.1 degrees Celsius (°C), the light wavelength can be built-in as an optional wavelength with an accuracy of nanometers (nm), and the gas pressure accuracy is Pascals (Pa).

[0079] In detail, the temperature control unit 210 includes a heating plate, a temperature sensor, and a first controller. The heating plate and temperature sensor are connected to the first controller. The heating plate is a commonly used laboratory electric heating plate accessory, used for heating under the control of the first controller. The temperature sensor is used to collect the temperature inside the sealed incubator 100 under the control of the first controller. In implementation, the rated power required by the heating plate and temperature sensor can be adjusted according to the specifications of the sealed incubator 100, which is not limited in this document.

[0080] The light control unit 220 includes a light-emitting device and a second controller. The light-emitting device is connected to the second controller and is used to generate light under the control of the second controller. Specifically, the light-emitting device can be a commonly used household color-changing lamp, with a built-in selectable wavelength that can be set to ultraviolet light, natural light, white light, and colored light of different wavelengths. When the light-emitting device is set to emit ultraviolet light, it is used to sterilize the interior of the sealed incubator 100. When the light-emitting device is set to other light sources, it is used to provide the light source required for microbial culture. In practice, multiple light emitters can also be selected, each emitting light of different wavelengths.

[0081] The pressure control unit 230 comprises a pressure regulating device, a pressure sensor and a third controller. Specifically, the pressure regulating device is configured to adjust the pressure in the closed incubator 100 under the control of the third controller, and the pressure sensor is configured to collect the pressure in the closed incubator 100 under the control of the third controller. In a specific implementation, in order to facilitate observation of the pressure change, the pressure sensor can also be arranged at a top corner position of the closed incubator 100.

[0082] In some embodiments, the temperature control unit 210, the light control unit 220 and the pressure control unit 230 can be controlled by respective controllers, or can be controlled by a unified controller, which is not limited herein.

[0083] In an embodiment, in order to improve the durability of the air inlet assembly 300 and the liquid inlet assembly 400, the material of the connecting pipeline of the air inlet assembly 300 and the liquid inlet assembly 400 is recommended to be an acid and alkali corrosion resistant polymer resin material, such as polyvinyl chloride (PVC), polyether ether ketone (PEEK), polytetrafluoroethylene (PTFE) and the like, and is not recommended to be an iron, steel or other corrosion resistant metal material or a plastic material.

[0084] The air inlet assembly 300 comprises a plurality of air inlet valves. In a specific implementation, in order to improve the air inlet speed and uniformity, the air inlet valves are arranged around the closed incubator, for example, on the top surface side wall and the side surface side wall, as shown in FIG. 4. Figure 3 In addition, in order to perform nitrogen charging, deoxidation and carbon dioxide saturation operations in the liquid environment in the closed incubator 100, at least two air inlet valves are arranged at the bottom of the closed incubator 100. The interface of the air inlet assembly 300 is a standard interface of a laboratory commonly used gas cylinder. In a specific implementation, a pressure gauge can be externally connected to control the air inlet pressure.

[0085] The liquid inlet assembly 400 comprises a plurality of liquid inlet valves arranged on the upper part or the side part of the outer wall of the closed incubator 100, so as to facilitate mixing of the solution and the liquid medium in the closed incubator 100 after input. The interface of the liquid inlet assembly 400 is a standard caliber commonly used in a laboratory, which can be connected to a liquid inlet pipeline through a tee joint or a rubber tube.

[0086] In a further embodiment, as shown in FIG. 5, the gas taking assembly 510 comprises a first valve 511, a second valve 512 and a gas taking chamber 513. Figure 5

[0087] The first valve 511 is arranged on the top outer wall of the closed incubator 100, the second valve 512 is provided with a gas taking port for connecting a gas sampler, and the gas taking chamber 513 is arranged between the first valve 511 and the second valve 512 and is configured to accommodate the sampled gas.

[0088] ​In detail, the first valve 511 and the second valve 512 can be selected as a spherical valve body, and other forms of valve body can also be selected, which is not limited herein. The first valve 511 is arranged close to the closed incubator 100, the second valve 512 is arranged away from the closed incubator 100, and the internal volume of the gas taking cabin 513 can be set according to requirements, which is not limited herein. In order to avoid gas leakage of the gas taking port in the second valve 512, a gas-tight pad is arranged on the gas taking port of the second valve 512.

[0089] In order to improve the service life of the gas taking assembly 510, the materials of the first valve 511, the second valve 512, the gas taking cabin 513 and the internal connecting pipe are recommended to use acid and alkali corrosion resistant high molecular resin materials such as polyvinyl chloride (PVC), polyether ether ketone (PEEK), polytetrafluoroethylene (PTFE) and the like, and iron, steel and other corrosion resistant metal materials and plastic materials are not recommended.

[0090] The gas taking process of the gas taking assembly 510 is as follows: opening the first valve 511 and closing the second valve 512 can make the gas in the closed incubator 100 enter the gas taking cabin 513; closing the first valve 511 and keeping the second valve 512 closed, the gas taking cabin 513 is connected with the gas taking port of the second valve 512 through the gas-tight pad, the gas sampling is performed and the subsequent test analysis is carried out; closing the first valve 511, opening the second valve 512, and connecting the second valve 512 with the vacuum pumping device, the gas in the gas taking cabin 513 is quickly removed by the vacuum pumping device, thereby avoiding cross contamination in the multiple gas taking processes.

[0091] In further embodiments of the present application, as shown in Figure 6A The liquid taking assembly 520 includes a first valve 521, a second valve 522 and a liquid taking cabin 523.

[0092] The first valve 521 is arranged at the bottom of the closed incubator 100; the second valve 522 is provided with a liquid taking port for connecting a liquid sampler; and the liquid taking cabin 523 is arranged between the first valve 521 and the second valve 522 for accommodating the sampled liquid.

[0093] In detail, the first valve 521 and the second valve 522 can be selected as a spherical valve body, and other forms of valve body can also be selected, which is not limited herein. The first valve 521 is arranged close to the closed incubator 100, the second valve 522 is arranged away from the closed incubator 100, and the internal volume of the liquid taking cabin 523 can be set according to requirements, which is not limited herein. In order to avoid gas leakage of the gas taking port in the second valve 522, a gas-tight pad is arranged on the gas taking port of the second valve 522.

[0094] To improve the service life of the liquid dispensing component 520, it is recommended that the materials of the first valve 521, the second valve 522, the liquid dispensing chamber 523, and the internal connecting pipe be made of acid and alkali resistant polymer resin materials, such as polyvinyl chloride (PVC), polyether ether ketone (PEEK), polytetrafluoroethylene (PTFE), etc. It is not recommended to use iron, steel, or other non-corrosion resistant metal materials and plastic materials.

[0095] The liquid collection process of the liquid collection component 520 includes: opening the first valve 521 and closing the second valve 522 to allow the liquid in the incubator to enter the liquid collection chamber 523; simultaneously closing the first valve 521 and the second valve 522, using a commonly used laboratory liquid sampler to pass through the airtight gasket of the sampling port of the second valve 522 to connect with the liquid collection chamber 523, aspirating the liquid for subsequent experimental analysis; and opening the second valve 522 to drain the liquid from the liquid collection chamber 523.

[0096] In practical implementation, to avoid cross-contamination from multiple liquid samplings, the liquid sampling assembly 520 also includes a third valve 524, such as... Figure 6B As shown, a cleaning solution such as deionized water or distilled water is introduced through a third valve 524 and discharged through the second valve 522, which can quickly remove the liquid in the liquid collection chamber 523 and achieve effective cleaning. In a specific implementation, the third valve 524 can be placed between the first valve 521 and the liquid collection chamber 523.

[0097] In further embodiments described herein, please refer to [link / reference]. Figure 4 The closed-loop microbial culture device also includes a microwave oscillation component 600, located at the bottom of the closed incubator 100, used to oscillate the liquid inside the closed incubator 100 (oscillation frequency accuracy is Hertz), so as to quickly integrate the liquid and energy inside the closed incubator 100, or to use the cleaning solution inside the closed incubator 100 to clean the inside of the closed incubator 100. Specifically, the rated power of the microwave oscillation component 600 can be adjusted according to the specifications of the closed incubator 100, which is not limited in this document.

[0098] In further embodiments described herein, please refer to [link / reference]. Figure 4 The closed microbial culture device also includes a support cabinet 700, which is located at the bottom of the closed incubator 100 to support the closed incubator 100 and to house the environmental control components 200.

[0099] In detail, the support cabinet 700 can be made of high-performance stainless steel (e.g., 304# stainless steel) or acid and alkali resistant polymer resin. The dimensions of the support cabinet 700 can be adjusted according to the specifications of the sealed incubator 100; this document does not impose any limitations on this. The support cabinet 700 incorporates a microwave oscillation component 600 and an environmental control component 200.

[0100] In further embodiments herein, reference is made to Figure 1 and Figure 2 As shown in FIG. 8, the closed microbial culture device further comprises a display and control assembly 800 disposed on the support cabinet 700, connected to the environmental control assembly 200 and the microwave oscillation assembly 600, for displaying the environmental parameters (including experimental temperature, light wavelength, gas pressure, etc.) and oscillation frequency in the closed incubator, and controlling the operation of the environmental control assembly 200.

[0101] In the formula, the precision of the oscillation frequency is Hertz (Hz), the precision of the temperature is 0.1 Celsius degree (℃), the light wavelength can be built-in as an optional wavelength with a precision of nanometer (nm), and the precision of the gas pressure is Pascal (Pa).

[0102] In specific implementation, the microbial culture device further comprises a power switch button 900 disposed on the support cabinet 700, one end of which is connected to the environmental control assembly 200 and the display and control assembly 800 through power lines of different specifications, and the other end of which is connected to a three-phase power plug. The power switch button 900 can be used to power on the environmental control assembly 200 and the display and control assembly 800.

[0103] Compared with the prior art, the microbial culture device provided in the present disclosure has at least the following advantages:

[0104] The microbial culture device can realize the culture of microorganisms in a specific closed environment, and can adjust five environmental factors including temperature, pressure, gas, water, temperature and light. In addition, the device can continuously realize the operations of gas feeding, liquid feeding, medium feeding, gas taking and liquid taking during the experiment, and can always maintain the isolation of the inside of the incubator from the outside environment, so as to ensure that the microbial culture process is not polluted by the outside environment, and to ensure the accuracy and continuity of the simulation experiment of the survival of microorganisms in a specific environment. The device has the advantages of easy operation, easy cleaning, easy disinfection, good sealing, high use efficiency, continuous experiment, and effective guarantee of the sampling process not being polluted by the outside environment.

[0105] In an embodiment herein, a microbial culture method suitable for the microbial culture device described in the foregoing embodiments is also provided, as shown in FIG. 7, comprising the following steps: Figure 7 As shown in FIG. 7, the method comprises the following steps:

[0106] In step 710, cleaning: a cleaning liquid is injected into the closed incubator through the liquid feeding assembly, and after cleaning, the cleaning liquid in the closed incubator is discharged through the liquid taking assembly.

[0107] In detail, the injection of the cleaning liquid into the closed incubator through the liquid feeding assembly comprises the following steps: opening the liquid feeding valve of the liquid feeding assembly and the gas taking valve of the gas taking assembly, closing other valves, and injecting the cleaning liquid through the liquid feeding valve. The discharge of the cleaning liquid through the liquid taking assembly comprises the following steps: opening the valve in the liquid taking assembly, while keeping the gas taking assembly open, so that the cleaning liquid is discharged through the valve in the liquid taking assembly.

[0108] The cleaning liquid used in this step is, for example, deionized water or distilled water. Specifically, in order to improve the cleaning degree, the microwave oscillation assembly is turned on after the cleaning liquid is filled, so as to perform microwave oscillation on the cleaning liquid. The cleaned liquid is discharged through the liquid taking assembly. In special cases, cleaning agents can also be added through other liquid inlet assemblies to enhance the cleaning effect. In addition, the cleaning effect can also be enhanced by repeatedly cleaning multiple times.

[0109] Before this step is implemented, the light control unit can also be turned on to generate ultraviolet light to irradiate the sealed incubator for a period of time, so as to perform sterilization treatment on the sealed incubator.

[0110] Step 720, liquid addition and gas removal: experimental liquid is injected into the sealed incubator through the liquid inlet assembly, and oxygen removal gas is injected into the sealed incubator through the gas inlet assembly, so as to perform oxygen removal treatment on the experimental liquid.

[0111] Specifically, the injection of experimental liquid into the sealed incubator through the liquid inlet assembly in this step includes: opening the valve of the liquid inlet assembly and the valve of the gas taking assembly, closing other valves, and injecting experimental liquid through the valve of the liquid inlet assembly.

[0112] The oxygen removal gas is, for example, nitrogen, which is not limited herein. In specific implementation, in order to balance the content of carbon dioxide in the sealed incubator, carbon dioxide is also injected after the oxygen removal gas is injected.

[0113] Step 730, gas filling: experimental gas is injected into the sealed incubator through the gas inlet assembly.

[0114] This step is opened when the experimental gas is different from the composition of the atmosphere on the earth at present. After different volumes and pressures of gas are injected and mixed, the injection of experimental gas is realized.

[0115] Step 740, medium addition: microbial culture medium is injected into the sealed incubator through the liquid inlet assembly.

[0116] After the above steps 710 to 740, all valves are closed.

[0117] Step 750, microbial culture: the environment in the sealed incubator is set through the environment control assembly (set the microwave oscillation frequency, temperature, and light source wavelength); and the microorganism is added into the sealed incubator through the liquid inlet assembly, and the microbial culture is started.

[0118] This embodiment can realize the culture of microorganisms in a specific sealed environment, and can adjust the environmental factors, gas and liquid factors in the sealed incubator. In addition, through the setting of the gas inlet assembly, the liquid inlet assembly and the liquid taking assembly, the sealed incubator can be cleaned without contact, so that repeated experiments can be realized.

[0119] Further, during the experiment, if the gas or liquid in the closed incubator is to be analyzed, the following method can be used to achieve the analysis, for example, as shown in Figure 8 The microbial culture method further includes:

[0120] At step 760, gas is taken out by the gas taking-out assembly to analyze the influence of the microbial activity on the gas according to the taken-out gas.

[0121] In detail, the gas in the closed incubator is taken out by the gas taking-out assembly, which includes: keeping other valves closed, and then opening the double valve in the gas taking-out assembly to take out the gas (the specific process of taking out the gas is described in the foregoing embodiment, which will not be described here again).

[0122] At step 770, liquid is taken out by the liquid taking-out assembly to analyze the influence of the microbial activity on the liquid according to the taken-out liquid.

[0123] In detail, the liquid in the closed incubator is taken out by the liquid taking-out assembly, which includes: keeping other valves closed, and then opening the double valve or the triple valve in the liquid taking-out assembly to take out the liquid (the specific process of taking out the liquid is described in the foregoing embodiment, which will not be described here again).

[0124] The present embodiment can realize the non-contact gas taking-out and liquid taking-out by the setting of the gas taking-out assembly and the liquid taking-out assembly, thereby avoiding the influence of the gas taking-out and liquid taking-out process on the culture environment.

[0125] It should be understood that, in various embodiments herein, the sequence of the above processes does not mean the execution sequence, and the execution sequence of the processes should be determined according to the functions and the inherent logic, and should not constitute any limitation on the implementation process of the embodiments herein.

[0126] It should also be understood that, in the embodiments herein, the term “and / or” only describes the association relationship of the associated objects, and means that there can be three relationships. For example, A and / or B can mean that A exists alone, A and B exist together, and B exists alone. In addition, the character “ / ” herein generally means that the front and rear associated objects are in an “or” relationship.

[0127] Those skilled in the art can realize that the units and algorithm steps of the examples described in combination with the embodiments disclosed herein can be realized by electronic hardware, computer software or a combination of both. In order to clearly illustrate the interchangeability of hardware and software, the components and steps of the examples have been described in the above description in general. Whether the functions are executed in hardware or software depends on the specific application and design constraints of the technical solution. A person skilled in the art can use different methods to implement the described functions for each specific application, but such implementation should not be considered beyond the scope of the present disclosure.

[0128] Those skilled in the art can clearly understand that, for the convenience and brevity of description, the specific working processes of the above-described system, device and unit can refer to the corresponding processes in the foregoing method embodiments, which will not be repeated here.

[0129] In several embodiments provided herein, it should be understood that the disclosed system, device and method can be implemented in other manners. For example, the above-described device embodiments are merely schematic, and the division of the units is merely a logical function division, and there can be another division manner in actual implementation, for example, a plurality of units or components can be combined or integrated into another system, or some features can be ignored or not executed. In addition, the displayed or discussed coupling or direct coupling or communication connection between the units can be indirect coupling or communication connection through some interfaces, devices or units, and can also be electrical, mechanical or other forms of connection.

[0130] The units described as separate components can or can not be physically separate, and the components shown as units can or can not be physical units, i.e., can be located in one place, or can be distributed on a plurality of network units. Some or all of the units can be selected according to actual needs to achieve the purpose of the embodiments provided herein.

[0131] In addition, each functional unit in each embodiment herein can be integrated into a processing unit, or each unit can exist physically, or two or more units can be integrated into one unit. The integrated unit can be realized in the form of hardware or in the form of a software functional unit.

[0132] The integrated unit, if realized in the form of a software functional unit and sold or used as an independent product, can be stored in a computer readable storage medium. Based on such understanding, the technical solutions herein, essentially or in the form of a contribution to the prior art, or all or part of the technical solutions can be embodied in the form of a software product. The computer software product is stored in a storage medium, and includes several instructions for causing a computer device (which can be a personal computer, a server, or a network device, etc.) to execute all or part of the steps of the methods described in each embodiment herein. The foregoing storage medium includes: a U disk, a mobile hard disk, a read-only memory (ROM, Read-Only Memory), a random access memory (RAM, Random Access Memory), a magnetic disk or an optical disk, and various media that can store program codes.

[0133] The principles and implementations of the present application are described in detail in the specific examples, and the above examples are only used to help understand the method and its core idea; meanwhile, for those skilled in the art, according to the idea of the present application, the specific implementation and application range will be changed, and the above description should not be understood as a limitation of the present application.

Claims

1. A closed microbial culture device, characterized by, The closed microbial culture device comprises a closed incubator, an environment control assembly, an air inlet assembly, a liquid inlet assembly, a microwave oscillation assembly, and a sampling assembly. The closed incubator is transparent and used for observing and culturing microorganisms. The environment control assembly is arranged at the bottom of the closed incubator and used for collecting and adjusting the environmental parameters in the closed incubator. The air inlet assembly is arranged on the outer wall of the closed incubator and used for providing air into the closed incubator. The liquid inlet assembly is arranged on the outer wall of the top of the closed incubator and used for providing liquid into the closed incubator. The sampling assembly is arranged on the outer wall of the closed incubator and used for extracting or discharging the gas or liquid in the closed incubator. The sampling assembly comprises a first valve, a second valve, and a sampling bin. The microwave oscillation assembly is arranged at the bottom of the closed incubator and used for cleaning the closed incubator by oscillating the cleaning liquid in the closed incubator after the closed incubator is filled with the cleaning liquid. The first valve is arranged on the outer wall of the closed incubator. The second valve is provided with a sampling port for connecting a sampling device. The sampling bin is arranged between the first valve and the second valve and used for containing the sampled gas or liquid.

2. The closed microbial culture device of claim 1, wherein The first valve is arranged close to the closed incubator, and the second valve is arranged away from the closed incubator.

3. The closed microbial culture device of claim 1, wherein The sampling process of the sampling assembly comprises the following steps: opening the first valve, closing the second valve, allowing the gas in the closed incubator to enter the sampling bin, closing the first valve, keeping the second valve closed, connecting the sampling bin with the air-tight pad of the sampling port of the second valve through a gas sampler, sampling the gas, and performing subsequent test analysis. closing the first valve, opening the second valve, connecting the second valve with a vacuum pump, and quickly removing the gas in the sampling bin by the vacuum pump to avoid cross contamination in the multiple sampling processes. The closed incubator is integrally formed by transparent plates or sealed by multiple transparent plates, and the bottom of the closed incubator is heat-conducting.

4. The closed microbial culture device of claim 1, wherein The environment control assembly comprises a temperature control unit, a light control unit, and a pressure control unit.

5. The closed microbial culture device of claim 1, wherein The temperature control unit is used for adjusting and measuring the temperature in the closed incubator.

6. The closed microbial culture device of claim 5, wherein The light control unit is used for adjusting and measuring the light in the closed incubator. The pressure control unit is used for adjusting and measuring the pressure in the closed incubator.

7. A method for culturing microorganisms, characterized by, For the sampling assembly for sampling liquid, a third valve is arranged between the first valve and the sampling bin or between the sampling bin and the second valve and used for inputting cleaning liquid to clean the sampling bin. A support cabinet is arranged at the lower part of the closed incubator and used for supporting the closed incubator and containing the environment control assembly. A display and control assembly is arranged on the support cabinet, connected with the environment control assembly, and used for displaying the environmental parameters in the closed incubator and controlling the operation of the environment control assembly. The closed microbial culture device is suitable for the closed microbial culture device in any one of claims 1 to 6. The liquid injection assembly injects cleaning liquid into the closed incubator, and the closed incubator is cleaned by oscillating the cleaning liquid in the closed incubator. After cleaning, the sampling assembly discharges the cleaning liquid in the closed incubator. The liquid injection assembly injects experimental liquid into the closed incubator, and the oxygen removal treatment of the experimental liquid is performed by injecting oxygen removal gas into the closed incubator through the gas injection assembly. The gas injection assembly injects experimental gas into the closed incubator. The liquid injection assembly injects microbial culture medium into the closed incubator. The environmental control assembly sets the environment in the closed incubator. The liquid injection assembly injects microorganisms into the closed incubator to start the culture of microorganisms. Also comprising: The sampling assembly takes gas by the following operations: Open the first valve, close the second valve, let the gas in the closed incubator enter the sampling chamber; close the first valve, keep the second valve closed, connect the gas sampling chamber with the airtight pad of the gas sampling port through the second valve, and take gas sample and perform subsequent test analysis; close the first valve, open the second valve, and connect the second valve with the vacuum device, and quickly remove the gas in the sampling chamber by the vacuum device, thereby avoiding cross contamination in the multiple gas sampling processes.

8. The microorganism culture method according to claim 7, wherein After starting the culture of microorganisms, further comprising: The sampling assembly extracts the liquid in the closed incubator to analyze the influence of the activity of microorganisms on the liquid according to the extracted liquid.

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