CN114907971A

CN114907971A - Closed microbial culture device and microbial culture method

Info

Publication number: CN114907971A
Application number: CN202110170700.5A
Authority: CN
Inventors: 王华建; 叶云涛; 张水昌; 王晓梅
Original assignee: Petrochina Co Ltd
Current assignee: Petrochina Co Ltd
Priority date: 2021-02-08
Filing date: 2021-02-08
Publication date: 2022-08-16

Abstract

Provided herein are a closed microbial culture apparatus and a microbial culture method, wherein the apparatus includes: the device comprises a closed incubator, an environment control assembly, an air inlet assembly, a liquid inlet assembly and a sampling assembly; the closed incubator is a transparent structure and is used for observing and culturing microorganisms; the environment control assembly is used for acquiring and adjusting environment parameters in the closed incubator; the gas inlet assembly is used for providing gas into the closed incubator; the liquid inlet component is used for providing liquid into the closed incubator; the sampling assembly is arranged on the outer wall of the closed incubator and is used for extracting or discharging gas or liquid in the closed incubator. The culture medium can realize the culture of microorganisms in a specific closed environment, can adjust environmental factors, gas and liquid factors in the closed incubator, and avoids the influence of gas and liquid taking processes on the culture environment. The device has the characteristics of convenient operation, easy cleaning, good tightness, high use efficiency and continuous experiment.

Description

Closed microbial culture device and microbial culture method

Technical Field

The invention relates to the field of microbial culture, in particular to a closed microbial culture device with adjustable environmental factors and a microbial culture method.

Background

Microorganisms are a large group of organisms including bacteria, viruses, fungi, some small protists, microalgae and the like, and the microorganisms are tiny in individuals and have great influence on the aspects of global environmental evolution, mineral resource formation, ecological environment change, industrial and agricultural production, human health and the like. Research related to microorganisms now involves many fields such as petroleum, mineral products, chemical industry, biology, medicine, food, industry and agriculture, and environmental protection. The cultivation of microorganisms in specific environments is also considered to be an important technical means of current scientific research.

The living environment of the microorganisms includes various environmental factors such as temperature, pressure, gas, water, illumination and the like, and the environmental factors directly influence the culture efficiency of the microorganisms. But these environmental factors may not be the same as the current earth environment. In order to correctly recognize the living environment and action mechanism of microorganisms, closed microorganism culture with adjustable environmental factors is necessary. Most of the existing microorganism incubators sold in the market are heating type constant temperature incubators, and the adjustable environmental factors are very limited. For example, the published Chinese patent with application number of 201810487432.8 and the name of constant temperature microbe incubator for avoiding cross contamination can be simply adjusted in environmental parameters such as temperature and illumination, but the parameters related to the living environment of microbes such as the atmosphere and water body can not be freely set.

Microbial culture is generally a continuous experiment within a certain time range, and requires multiple sets of comparisons to grasp growth curves of microbes under different environmental factors and screen out the most favorable environmental factors. The existing closed microorganism incubator mostly provides a box space for placing a microorganism culture dish, and is not a direct culture space. Consequently constantly open at the incubator, the sample in-process, the atmosphere and the water environment of culture dish can receive the interference, easily cause cross contamination or cultivate the environment out of control, and then influence the result of cultivateing. In order to avoid cross contamination, experimenters often need to culture multiple microorganism samples under the same environmental element. Therefore, the process of the microorganism culture experiment with special environmental elements is extremely complicated, and the strict consistency of all the set parameters of the parallel samples with the same environmental elements is difficult to ensure. In addition, the number of the used microorganism incubators is exponentially increased in the experimental process, but the efficiency is low.

Disclosure of Invention

The microbial incubator is used for solving the problems that the microbial incubator in the prior art has limited adjustable environmental factors, low use efficiency and is not beneficial to microbial culture.

In order to solve the above technical problems, a first aspect of the present disclosure provides a closed microbial cultivation apparatus, including: the device comprises a closed incubator, an environment control assembly, an air inlet assembly, a liquid inlet assembly and a sampling assembly;

the closed incubator is of a transparent structure and is used for observing and culturing microorganisms;

the environment control assembly is arranged at the bottom of the closed incubator and is used for collecting and adjusting environment parameters in the closed incubator;

the air inlet assembly is arranged on the outer wall of the closed incubator and is used for providing air into the closed incubator;

the liquid inlet component 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 is used for extracting or discharging gas or liquid in the closed incubator; the sampling assembly comprises a first valve, a second valve and a sampling bin;

the first valve is arranged on the outer wall of the closed incubator; the second valve is provided with a sampling port which is used for communicating with sampling equipment; the sampling bin is arranged between the first valve and the second valve and used for containing sampled gas or liquid.

In a further embodiment of the invention, the closed incubator is formed by integrally molding a transparent plate or sealing a plurality of transparent plates, and the bottom of the closed incubator can conduct heat.

In a further embodiment herein, the environmental control assembly comprises: the device comprises a temperature control unit, a light control unit and a pressure control unit;

the temperature control unit is used for adjusting and measuring the temperature in the closed incubator;

the light control unit is used for adjusting and measuring illumination in the closed incubator;

the pressure control unit is used for adjusting and measuring the pressure in the closed culture box.

In a further embodiment herein, the sampling assembly comprises a gas extraction assembly and a liquid extraction assembly, wherein the liquid extraction assembly further comprises: and the 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 so as to clean the sampling bin.

In a further embodiment herein, the enclosed microbial cultivation apparatus further comprises: and the microwave oscillation assembly is arranged at the bottom of the closed incubator and is used for uniformly mixing the liquid in the closed incubator through microwave oscillation.

In a further embodiment herein, the enclosed microbial cultivation apparatus further comprises: and the supporting cabinet is arranged at the lower part of the closed incubator, is used for supporting the closed incubator and accommodating the environment control assembly.

In a further embodiment herein, the enclosed microbial cultivation apparatus further comprises: and the display control assembly is arranged on the supporting cabinet and connected with the environment control assembly and used for displaying the environmental parameters in the closed incubator and controlling the environment control assembly to work.

In a second aspect, there is provided a microorganism culture method, which is suitable for use in the enclosed microorganism culture apparatus of any one of the preceding embodiments, and comprises:

injecting cleaning liquid into the closed incubator through the liquid inlet assembly, and discharging the cleaning liquid in the closed incubator through the sampling assembly after cleaning;

injecting experimental liquid into the closed incubator through the liquid inlet assembly, and injecting oxygen-driving gas into the closed incubator through the air inlet assembly so as to perform oxygen removal treatment on the experimental liquid;

injecting experimental gas into the closed incubator through the gas inlet assembly;

injecting a microorganism culture medium into the closed incubator through a liquid inlet assembly;

setting the environment in the closed incubator through an environment control assembly;

and adding microorganisms into the closed incubator through a liquid inlet assembly to start culturing the microorganisms.

In further embodiments herein, after the beginning of culturing the microorganism, the method further comprises:

and extracting gas in the closed incubator through the gas extraction assembly so as to analyze the influence of microbial activity on the gas according to the extracted gas.

In further embodiments herein, after the beginning of culturing the microorganism, further comprising:

and extracting the liquid in the closed culture box through the liquid extracting assembly so as to analyze the influence of the microbial activity on the liquid according to the extracted liquid.

According to the closed microorganism culture device and the microorganism culture method, the culture of microorganisms in a specific closed environment can be realized through the arrangement of the closed culture box, the environment control assembly, the air inlet assembly and the liquid inlet assembly, and the environmental factors in the closed culture box and the gas and liquid factors can be adjusted. Through the setting of sampling assembly, can realize that the contactless gets gas and gets liquid, avoid getting gas and getting the influence of liquid process to cultivateing the environment. In addition, through the setting of subassembly, feed liquor subassembly and sampling subassembly of admitting air, can realize airtight incubator's contactless washing to realize the repetition test. The device has the characteristics of convenient operation, easy cleaning, good tightness, high use efficiency and continuous experiment, and can be applied to the fields of petroleum, mineral products, chemical industry, biology, medicine, food, industry and agriculture, environmental protection and the like.

In order to make the aforementioned and other objects, features and advantages of the present invention comprehensible, preferred embodiments accompanied with figures are described in detail below.

Drawings

In order to more clearly illustrate the embodiments or technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a schematic perspective view of a closed microorganism culture apparatus according to an embodiment of the present invention;

FIG. 2 is a schematic front view of a closed microbial cultivation apparatus according to an embodiment of the present invention;

FIG. 3 is a schematic top view of a closed microbial cultivation apparatus according to an embodiment of the present invention;

FIG. 4 is a schematic side view of a closed microbial cultivation apparatus according to an embodiment of the present invention;

FIG. 5 illustrates a side view schematic of an air extraction assembly according to embodiments herein;

FIGS. 6A and 6B are schematic side views of a fluid extraction assembly according to embodiments herein;

FIG. 7 shows a first flowchart of a microorganism cultivation method according to the embodiment herein;

FIG. 8 shows a second flowchart of a microorganism culturing method of the embodiments herein.

Description of the symbols of the drawings:

100. sealing the incubator;

200. an environmental control component;

300. an air intake assembly;

400. a liquid inlet component;

500. a sampling assembly;

510. a gas taking assembly;

520. a liquid taking assembly;

210. a temperature control unit;

220. a light control unit;

230. a voltage control unit;

511. a first valve;

512. a second valve;

513. a gas taking bin;

521. a first valve;

522. a second valve;

523. a liquid taking bin;

524. a third valve;

600. a microwave oscillating assembly;

700. a support cabinet;

800. a display control component;

900. a power switch button.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments herein without making any creative effort, shall fall within the scope of protection.

The microorganism incubator in the prior art is only a box space for placing a microorganism culture dish, and cannot realize the arrangement of microorganism culture environment, gas, liquid and the like. In addition, the microorganism incubator in the prior art is opened to sample, so that cross contamination or environmental runaway are easily caused, and the culture effect is further influenced.

In order to solve the above technical problems in the prior art, in one embodiment, there is provided a totally enclosed microbial cultivation apparatus, as shown in fig. 1, comprising: closed incubator 100, environmental control subassembly 200, admit air subassembly 300, inlet means 400, sampling subassembly 500.

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

The environmental control assembly 200 is disposed at the bottom of the closed incubator 100 and is used for collecting and adjusting environmental parameters in the closed incubator 100.

The gas inlet module 300 is disposed on an outer wall of the closed container 100, and supplies gas into the closed container 100.

The liquid inlet assembly 400 is provided on the top or side wall of the closed cultivation box 100 for supplying liquid into the closed cultivation box 100.

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

In particular, to avoid interference between gas and liquid sampling, the sampling assembly 500 includes a gas-taking assembly and a liquid-taking assembly. Wherein, get the gas subassembly and set up on airtight incubator 100 top outer wall for draw or discharge the gas in airtight incubator 100. The liquid taking component is arranged on the outer wall of the bottom of the closed incubator 100 and is used for taking or discharging liquid in the closed incubator 100.

The airtight microbial cultivation device that this embodiment provided, through airtight incubator, environmental control subassembly, air intake subassembly, feed liquor subassembly's setting, can realize the cultivation of microorganism under specific airtight environment, the environmental factor in the adjustable airtight incubator to and gaseous and liquid factor. Through getting the setting of gas subassembly and getting the liquid subassembly, can realize that the contactless gets gas and get liquid, avoid getting gas and getting the influence of liquid process to cultivateing the environment. In addition, through the setting of subassembly, feed liquor subassembly and the subassembly of getting liquid of admitting air, can realize the contactless washing of airtight incubator to realize the repetition test. The device has the characteristics of convenient operation, easy cleaning, good tightness, high use efficiency and continuous experiment, and can be applied to the fields of petroleum, mineral products, chemical industry, biology, medicine, food, industry and agriculture, environmental protection and the like.

In one embodiment of the present invention, in order to facilitate observing the microorganism culturing process, the closed incubator 100 is made of a transparent plate (e.g. organic glass), and specifically, the closed incubator can be integrally formed by the transparent plate, or formed by sealing and gluing a plurality of transparent plate plates, and the specific implementation form is not limited herein. Further, in order to improve the heating capacity of the closed cultivation box 100, the bottom of the closed cultivation box 100 may conduct heat. In order to ensure that the sealed incubator 100 is not easily broken, the thickness of the sealed incubator is, for example, about 12 mm.

In one embodiment, as shown in fig. 2, 3 and 4, the environmental control assembly 200 includes: a temperature control unit 210, a light control unit 220, and a voltage control unit 230;

the temperature control unit 210 is used to adjust and measure the temperature inside the closed incubator 100. The light control unit 220 is used for adjusting and measuring the illumination in the closed incubator 100. The pressure control unit 230 is used to adjust and measure the pressure inside the closed incubator 100. Wherein the temperature precision is 0.1 degree centigrade (DEG C), the lamp light wavelength can be built in the selectable wavelength with the precision of nanometer (nm), and the gas pressure precision is Pascal (Pa).

In detail, the temperature control unit 210 includes a heating plate, a temperature sensor, and a first controller. Heating plate and temperature sensor connect first controller, and the heating plate is the laboratory and uses electrical heating board accessory commonly for heating under the control of first controller, and temperature sensor is used for gathering the temperature in airtight incubator 100 under the control of first controller. In practice, the rated power required for the heating plate and the temperature sensor can be adjusted according to the specification of the closed incubator 100, which is not limited herein.

The light control unit 220 includes a light emitting device and a second controller, wherein the light emitting device is connected to the second controller for generating light under the control of the second controller. Specifically, the light-emitting device can be a household common color-changing lamp tube, the lamp light wavelength is a built-in selectable item, and ultraviolet light, natural light, white light and colored lamp light with different wavelengths can be set. The light emitting device is configured to emit ultraviolet light for sterilization of the inside of the closed cultivation box 100, and is configured to provide a light source required for culturing microorganisms when other light is emitted. In practice, the light emitting device may further comprise a plurality of light emitters for emitting light of different wavelengths.

The pressure control unit 230 includes a pressure adjusting device, a pressure sensor, and a third controller. Specifically, the pressure regulating device is used for regulating the pressure in the closed cultivation box 100 under the control of the third controller, and the pressure sensor is used for collecting the pressure in the closed cultivation box 100 under the control of the third controller. In particular, in order to observe the pressure variation, the pressure sensor may be disposed at a corner of the top of the closed cultivation box 100.

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, and in other embodiments, the temperature control unit, the light control unit 220, and the pressure control unit can also be controlled by a unified controller, which is not limited herein.

In one embodiment, in order to improve the durability of the air inlet assembly 300 and the liquid inlet assembly 400, the connecting pipes of the air inlet assembly 300 and the liquid inlet assembly 400 are made of a polymer resin material resistant to acid and alkali corrosion, such as polyvinyl chloride (PVC), Polyetheretherketone (PEEK), Polytetrafluoroethylene (PTFE), and the like, and iron, steel, and other metal materials and plastic materials which are not corrosion-resistant are not recommended.

The air inlet assembly 300 includes a plurality of air inlet valves, and in one embodiment, is disposed around the closed cultivation box, such as the top side wall and the side wall, as shown in fig. 3, in order to improve the air inlet speed and the uniformity of air inlet. In addition, at least two air intake valves are disposed at the bottom of the closed incubator 100 for the nitrogen and oxygen charging, carbon dioxide saturation, and other operations of the liquid environment in the closed incubator 100. The interface of the air inlet assembly 300 is a standard interface of a common gas cylinder in a laboratory, and in specific implementation, the air inlet assembly can be externally connected with a partial pressure gauge to control air inlet pressure.

The liquid inlet assembly 400 comprises a plurality of liquid inlet valves which are arranged on the outer wall of the upper part or the lateral part of the closed incubator 100, so that the solution and the liquid culture medium in the closed incubator 100 can be uniformly mixed after being input, and the interface of the liquid inlet assembly 400 is of a standard caliber commonly used in a laboratory and can be connected with a liquid inlet pipeline through a tee joint or a rubber pipe.

In further embodiments herein, as shown in fig. 5, the air extraction assembly 510 comprises: a first valve 511, a second valve 512 and a gas extraction bin 513.

The first valve 511 is arranged on the outer wall of the top of the closed incubator 100; the second valve 512 is provided with an air intake port which is used for communicating the gas sampler; the gas sampling bin 513 is disposed between the first valve 511 and the second valve 512, and is used for containing sampled gas.

In detail, the first valve 511 and the second valve 512 may be, for example, ball-shaped valve bodies, or may be other valve bodies, which is not limited herein. First valve 511 is disposed near the closed incubator 100, and second valve 512 is disposed far from the closed incubator 100, and the internal volume of the gas extraction chamber 513 can be set according to the requirement, which is not limited herein. To avoid air leakage through the air intake port of the second valve 512, an air-tight gasket is provided on the air intake port of the second valve 512.

In order to prolong the service life of the gas extraction assembly 510, the first valve 511, the second valve 512, the gas extraction chamber 513 and the internal connection pipe are made of polymer resin materials resistant to acid and alkali corrosion, such as polyvinyl chloride (PVC), Polyetheretherketone (PEEK), Polytetrafluoroethylene (PTFE), etc., and iron, steel, other metal materials and plastic materials which are not corrosion-resistant are not recommended.

The gas extraction process of the gas extraction assembly 510 is as follows: the first valve 511 is opened, the second valve 512 is closed, and the gas in the closed incubator 100 can enter the gas taking bin 513; closing the first valve 511, keeping the second valve 512 closed, communicating the gas sampling device with the gas sampling chamber 513 through the airtight pad of the gas taking port of the second valve 512, and performing gas suction sampling and subsequent experimental analysis; the first valve 511 is closed, the second valve 512 is opened, the second valve 512 is connected with a vacuumizing device, and the vacuumizing device quickly removes the gas in the gas taking bin 513, so that cross contamination in multiple gas taking processes is avoided.

In further embodiments herein, as shown in fig. 6A, the liquid extraction assembly 520 comprises: a first valve 521, a second valve 522 and a liquid taking chamber 523.

The first valve 521 is arranged at the bottom of the closed incubator 100; a liquid taking port is formed in the second valve 522 and is used for communicating the liquid sampler; the liquid taking chamber 523 is disposed between the first valve 521 and the second valve 522 for accommodating the sampled liquid.

In detail, the first valve 521 and the second valve 522 may be, for example, spherical valve bodies, or may be other valve bodies, which is not limited herein. The first valve 521 is disposed near the closed cultivation box 100, the second valve 522 is disposed far from the closed cultivation box 100, and the internal volume of the liquid taking chamber 523 can be set according to the requirement, which is not limited herein. To avoid air leakage from the air intake port in the second valve 522, an air-tight gasket is provided on the air intake port of the second valve 522.

In order to prolong the service life of the liquid taking assembly 520, the materials of the first valve 521, the second valve 522, the liquid taking chamber 523 and the internal connecting pipe are preferably high molecular resin materials resistant to acid and alkali corrosion, such as polyvinyl chloride (PVC), Polyetheretherketone (PEEK), Polytetrafluoroethylene (PTFE) and the like, and iron, steel, other metal materials and plastic materials which are not corrosion-resistant are not preferably used.

The liquid extraction process of the liquid extraction assembly 520 comprises: the first valve 521 is opened, the second valve 522 is closed, and the liquid in the incubator can enter the liquid taking bin 523; simultaneously closing the first valve 521 and the second valve 522, penetrating an airtight pad of a sampling port of the second valve 522 by using a common laboratory liquid sampler to be communicated with the liquid taking bin 523, sucking liquid and sampling, and performing subsequent experimental analysis; the second valve 522 is opened to discharge the liquid in the liquid taking chamber 523.

In specific implementation, in order to avoid cross contamination of multiple liquid taking operations, the liquid taking assembly 520 further includes a third valve 524, as shown in fig. 6B, the third valve 524 is disposed between the liquid taking chamber 523 and the second valve 522, and a cleaning solution such as deionized water or distilled water is input through the third valve 524 and discharged through the second valve 522, so that the liquid in the liquid taking chamber 523 can be quickly removed, and effective cleaning is achieved. In one embodiment, the third valve 524 may be disposed between the first valve 521 and the liquid-taking chamber 523.

In a further embodiment herein, referring again to fig. 4, the enclosed microbial cultivation apparatus further comprises: the microwave oscillating assembly 600 is disposed at the bottom of the closed incubator 100, and is configured to oscillate the liquid in the closed incubator 100 (the oscillation frequency is hz) so as to rapidly integrate the liquid and energy in the closed incubator 100, or to clean the closed incubator 100 by using the cleaning liquid in the closed incubator 100. Specifically, the power rating of the microwave oscillating assembly 600 may be adjusted according to the specification of the closed cultivation box 100, which is not limited herein.

In a further embodiment herein, referring again to fig. 4, the enclosed microbial cultivation apparatus further comprises: and a support cabinet 700 disposed at a lower portion of the closed cultivation box 100, for supporting the closed cultivation box 100 and accommodating the environment control assembly 200.

In detail, the supporting cabinet 700 may be made of high performance stainless steel (for example, 304# stainless steel), and a polymer resin material resistant to acid and alkali corrosion. The size of the supporting cabinet 700 may be adjusted according to the size of the closed cultivation box 100, which is not limited herein. The supporting cabinet 700 is internally provided with a microwave oscillation assembly 600 and an environment control assembly 200.

In a further embodiment, referring to fig. 1 and 2, the enclosed microbial cultivation apparatus further comprises: and the display control assembly 800 is arranged on the supporting cabinet 700, is connected with the environment control assembly 200 and the microwave oscillation assembly 600, and is used for displaying environmental parameters (including experiment temperature, light wavelength, gas pressure and the like) and oscillation frequency in the closed incubator and controlling the environment control assembly 200 to work.

Wherein, the precision of oscillation frequency is Hertz (Hz), the precision of temperature is 0.1 centigrade (DEG C), the light wavelength can be embedded with the selectable wavelength with the precision of nanometer (nm), and the precision of gas pressure is Pascal (Pa).

In particular, the microorganism culture apparatus further comprises: the power switch button 900 is arranged on the supporting cabinet 700, one end of the power switch button is connected with the environment control assembly 200 and the display control assembly 800 through power lines of different specifications, the other end of the power switch button is connected with a three-phase power plug, and the environment control assembly 200 and the display control assembly 800 can be powered on through the power switch button 900.

Compared with the prior art, the microorganism culture device provided by the invention has at least the following advantages:

the culture of microorganisms under specific closed environment can be realized, 5 environmental factors of temperature, pressure, gas, water, temperature and illumination can be adjusted, operations such as air entrainment, liquid feeding, culture medium addition, gas taking and liquid taking can also be realized continuously in the experimental process, the isolation between the inside of the incubator and the external environment is kept all the time, the microorganism culture process is prevented from being polluted by the external environment, and the accuracy and the continuity of microorganism survival simulation experiment under the specific environment are ensured. The device has the advantages of convenient operation, easy cleaning and disinfection, good sealing property, high use efficiency, continuous experiment and capability of effectively ensuring that the sampling process is not polluted by the external environment.

In an embodiment of the present disclosure, there is also provided a method for culturing microorganisms, as shown in fig. 7, adapted to the foregoing embodiment, including:

step 710, cleaning: injecting cleaning liquid into the closed incubator through the liquid inlet assembly, and discharging the cleaning liquid in the closed incubator through the liquid taking assembly after cleaning.

In detail, this step is passed through the feed liquor subassembly and is injected washing liquid including into airtight incubator into: and opening a liquid inlet valve of the liquid inlet assembly and a gas taking valve of the gas taking assembly, closing other valves, and injecting cleaning liquid through the liquid inlet valve. This step includes through getting liquid subassembly discharge washing liquid: the valve in the liquid take-off assembly is opened while the gas take-off assembly is kept open, thereby draining the cleaning liquid through the valve of the liquid take-off assembly.

The cleaning liquid used in this step is, for example, deionized water or distilled water, specifically, in order to improve the cleanliness, after the cleaning liquid is filled, the microwave oscillation assembly is opened to perform microwave oscillation on the cleaning liquid, and the liquid after cleaning is discharged through the liquid taking assembly. Under the special circumstances, can also add the cleaner in order to strengthen the cleaning performance through other feed liquor subassemblies, in addition, still can increase the cleaning performance through the mode of wasing many times repeatedly.

Before the step is implemented, the sealed incubator can be sterilized by opening the light control unit to generate ultraviolet light to irradiate the sealed incubator for a period of time.

Step 720, liquid adding and degassing: liquid for experiments is injected into the closed incubator through the liquid inlet assembly, and oxygen driving gas is injected into the closed incubator through the air inlet assembly so as to carry out deoxidization treatment on the liquid for experiments.

In detail, the step of injecting the experimental liquid into the closed incubator through the liquid inlet assembly comprises the following steps: and opening the valve of the liquid inlet assembly and the valve of the gas taking assembly, closing other valves, and injecting the experimental liquid through the valve of the liquid inlet assembly.

The oxygen-scavenging gas is, for example, nitrogen, and is not limited herein. In specific implementation, in order to balance the content of carbon dioxide in the closed incubator, carbon dioxide is injected after the oxygen-scavenging gas is injected.

Step 730, fill gas: injecting experimental gas into the closed incubator through the gas inlet assembly.

The method is started when the experimental gas is different from the existing atmosphere on the earth, and the experimental gas injection is realized by injecting the gas with different volumes and pressures and uniformly mixing.

Step 740, adding culture medium: injecting a microorganism culture medium into the closed incubator through the liquid inlet component.

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

Step 750, culturing the microorganisms: setting the environment in the closed incubator (setting the microwave oscillation frequency, the temperature and the wavelength of the light source) through an environment control component; and adding microorganisms into the closed incubator through the liquid inlet assembly to start culturing the microorganisms.

The embodiment can realize the culture of microorganisms in a specific closed environment, and can adjust the environmental factors in the closed incubator and the factors of gas and liquid. In addition, through the setting of subassembly, feed liquor subassembly and the subassembly of getting liquid of admitting air, can realize the contactless washing of airtight incubator to realize the repetition test.

Further, in the experiment process, if it is desired to analyze the gas or liquid in the closed incubator, the method may be implemented as follows, as shown in fig. 8, and the method further includes:

step 760, gas taking: and extracting gas in the closed incubator by the gas extraction assembly so as to analyze the influence of the microbial activity on the gas according to the extracted gas.

In detail, the gas extraction from the closed incubator by the gas extraction assembly comprises: keeping other valves closed, gas extraction is performed by opening double valves in the gas extraction assembly (the gas extraction process is described in the previous embodiment and will not be described in detail here).

Step 770, taking liquid: and extracting the liquid in the closed incubator by the liquid extracting component so as to analyze the influence of the microbial activity on the liquid according to the extracted liquid.

In detail, the extraction of the liquid in the closed incubator by the liquid extraction assembly comprises: taking liquid through a double valve or a triple valve in the liquid taking assembly under the condition that other valves are kept closed (the liquid taking process is specifically referred to the previous embodiment and is not described in detail here).

This embodiment can realize that the contactless gets gas and get liquid through getting the setting of gas subassembly and getting the liquid subassembly, avoids getting gas and getting the influence of liquid process to cultivateing the environment.

It should be understood that, in various embodiments herein, the sequence numbers of the above-mentioned processes do not mean the execution sequence, and the execution sequence of each process should be determined by its function and inherent logic, and should not constitute any limitation to the implementation process of the embodiments herein.

It should also be understood that, in the embodiments herein, the term "and/or" is only one kind of association relation describing an associated object, meaning that three kinds of relations may exist. For example, a and/or B, may represent: a exists alone, A and B exist simultaneously, and B exists alone. In addition, the character "/" herein generally indicates that the former and latter associated objects are in an "or" relationship.

Those of ordinary skill in the art will appreciate that the elements and algorithm steps of the examples described in connection with the embodiments disclosed herein may be embodied in electronic hardware, computer software, or combinations of both, and that the components and steps of the examples have been described in a functional general in the foregoing description for the purpose of illustrating clearly the interchangeability of hardware and software. Whether such functionality is implemented as hardware or software depends upon the particular application and design constraints imposed on the technical solution. Skilled artisans may implement the described functionality in varying ways for each particular application, but such implementation decisions should not be interpreted as causing a departure from the scope of the present disclosure.

It is clear to those skilled in the art that, for convenience and brevity of description, the specific working processes of the above-described systems, apparatuses and units may refer to the corresponding processes in the foregoing method embodiments, and are not described herein again.

In the several embodiments provided herein, it should be understood that the disclosed system, apparatus, and method may be implemented in other ways. For example, the above-described apparatus embodiments are merely illustrative, and for example, the division of the units is only one logical division, and other divisions may be realized in practice, for example, a plurality of units or components may be combined or integrated into another system, or some features may be omitted, or not executed. In addition, the shown or discussed mutual coupling or direct coupling or communication connection may be an indirect coupling or communication connection through some interfaces, devices or units, and may also be an electric, mechanical or other form of connection.

The units described as separate parts may or may not be physically separate, and parts displayed as units may or may not be physical units, may be located in one place, or may be distributed on a plurality of network units. Some or all of the units can be selected according to actual needs to achieve the purposes of the embodiments herein.

In addition, functional units in the embodiments herein may be integrated into one processing unit, or each unit may exist alone physically, or two or more units are integrated into one unit. The integrated unit may be implemented in the form of hardware, or may also be implemented in the form of a software functional unit.

The integrated unit, if implemented in the form of a software functional unit and sold or used as a stand-alone product, may be stored in a computer readable storage medium. Based on such understanding, the technical solutions of the present invention may be implemented in a form of a software product, which is stored in a storage medium and includes several instructions for causing a computer device (which may be a personal computer, a server, or a network device) to execute all or part of the steps of the methods described in the embodiments of the present invention. And the aforementioned storage medium includes: a U-disk, a removable hard disk, a Read-Only Memory (ROM), a Random Access Memory (RAM), a magnetic disk, or an optical disk, and various media capable of storing program codes.

The principles and embodiments of the present disclosure are explained in detail by using specific embodiments, and the above description of the embodiments is only used to help understanding the method and its core idea; meanwhile, for the general technical personnel in the field, according to the idea of this document, there may be changes in the concrete implementation and the application scope, in summary, this description should not be understood as the limitation of this document.

Claims

1. An enclosed microbial culture apparatus, comprising: the device comprises a closed incubator, an environment control assembly, an air inlet assembly, a liquid inlet assembly and a sampling assembly;

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;

2. The enclosed microbial cultivation apparatus according to claim 1, wherein the enclosed incubator is formed by integrally molding a transparent plate or sealing a plurality of transparent plates, and the bottom of the enclosed incubator is heat conductive.

3. The enclosed microbial culture apparatus of claim 1, wherein the environmental control assembly comprises: the device comprises a temperature control unit, a light control unit and a pressure control unit;

4. The closed microbe culture apparatus of claim 1, wherein the sampling assembly for liquid extraction further comprises: and the 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 so as to clean the sampling bin.

5. The enclosed microbial culture apparatus of claim 1, further comprising: and the microwave oscillation assembly is arranged at the bottom of the closed incubator and is used for uniformly mixing the liquid in the closed incubator through microwave oscillation.

6. The enclosed microbial culture apparatus of claim 1, further comprising: and the supporting cabinet is arranged at the lower part of the closed incubator, is used for supporting the closed incubator and accommodating the environment control assembly.

7. The enclosed microbial culture apparatus of claim 6, further comprising: and the display control assembly is arranged on the supporting cabinet and connected with the environment control assembly and used for displaying the environmental parameters in the closed incubator and controlling the environment control assembly to work.

8. A microorganism culture method applied to the closed microorganism culture apparatus according to any one of claims 1 to 7, comprising:

injecting experimental liquid into the closed incubator through the liquid inlet assembly, and injecting oxygen-expelling gas into the closed incubator through the gas inlet assembly so as to perform oxygen removal treatment on the experimental liquid;

9. The method of claim 8, further comprising, after the beginning of culturing the microorganism:

and extracting gas in the closed incubator through the sampling assembly so as to analyze the influence of the microbial activity on the gas according to the extracted gas.

10. The method of claim 8, further comprising, after the beginning of culturing the microorganism:

and extracting the liquid in the closed culture box through the sampling assembly so as to analyze the influence of the microbial activity on the liquid according to the extracted liquid.