Microorganism culture system

Abstract

The utility model provides a microorganism culture system which comprises at least one set of subsystems. Wherein the subsystem comprises a culture container, a sampling container, a first container, a second container, a third container, a liquid storage ring, an injection pump, a switching valve with a public channel opening and a plurality of channel openings, a culture medium providing device and a liquid level maintaining device. The injection pump is matched with the switching valve, so that liquid in each container can be sucked; the culture medium providing device can provide a culture medium for the culture container; when the liquid level in the culture container exceeds the preset height, the liquid level maintaining device can suck away redundant liquid so that the liquid level in the culture container does not exceed the preset height. It can be seen that liquid suction, culture medium providing and microbial solution volume maintaining can be achieved through the microbial culture system, manual operation of a user is not needed, and efficiency is improved.

Description

Technical Field

[0001] The utility model relates to the field of microbial cultivation, in particular to a microbial cultivation system. Background Art

[0002] Existing microbial cultivation is all done manually. The operator places the bacteria to be evolved on a shaker, waits for a while (the growth rate of bacteria is different in different experimental environments), takes out the bacteria and measures its OD value (absorbance). The OD value reflects the concentration of the bacteria. When the OD value is greater than a certain value, a small amount of the original solution is taken out and the culture medium is added, and then the shaker is put back for evolution. This method is time-consuming, labor-intensive, and extremely inefficient.

[0003] Therefore, the efficiency of existing microbial culture needs to be improved. Utility Model Content

[0004] The present application provides a microorganism culture system, aiming to improve the efficiency of microorganism culture.

[0005] One embodiment provides a microorganism cultivation system, comprising at least one subsystem; the subsystem comprises:

[0006] Culture vessels, used to provide a place for microorganisms to evolve;

[0007] A sampling container, used for holding the sample solution drawn from the culture container by the syringe pump;

[0008] A first container for containing a growth inhibitor;

[0009] A third container is used to hold a cleaning solution;

[0010] Liquid storage ring;

[0011] Syringe pump;

[0012] A switching valve having a common channel port and a plurality of channel ports, wherein the injection pump is connected to the common channel port of the switching valve through the liquid storage ring, and the culture container, the sampling container, the first container and the third container are respectively connected to different channel ports of the switching valve; the switching valve is used to selectively connect one of the plurality of channel ports to the common channel port;

[0013] A culture medium supply device, used for supplying culture medium to the culture container;

[0014] The liquid level maintaining device is used to suck away excess liquid when the liquid level in the culture container exceeds a preset height so that the liquid level in the culture container does not exceed the preset height.

[0015] In the system provided by one embodiment, the liquid level maintaining device is further used to suck external air into the culture container when the liquid level in the culture container does not exceed a preset height.

[0016] In the system provided by one embodiment, the liquid level maintaining device comprises:

[0017] A vacuum pump; the suction end of the vacuum pump is connected to the culture container through a first pipette; the first pipette extends into the culture container, and the height of the bottom end of the first pipette is the preset height.

[0018] In the system provided by one embodiment, the liquid level maintaining device further comprises:

[0019] A water storage container is used for storing water; the water storage container is connected to the culture container; an air inlet pipe is arranged in the water storage container, one end of the air inlet pipe is connected to the atmosphere through a filter component, and the other end of the air inlet pipe extends below the water surface.

[0020] In the system provided by an embodiment, the subsystem further comprises a waste liquid container, the waste liquid container is connected to a channel port of the switching valve and is also connected to a discharge end of the vacuum pump;

[0021] The liquid level maintaining device also includes a three-way valve; the suction end of the vacuum pump is connected to the outlet end of the three-way valve, the first inlet end of the three-way valve is connected to the culture container through a first pipette, the second inlet end of the three-way valve is connected to the culture container through a second pipette, and the second pipette extends into the bottom of the culture container; the three-way valve selectively connects one of the first inlet end and the second inlet end to the outlet end.

[0022] In the system provided in one embodiment, the subsystem also includes a fourth container for holding an isolation liquid, and the isolation liquid is used to isolate the liquid and air in other containers; the fourth container is connected to a channel port of the switching valve; the injection pump sucks the isolation liquid in the fourth container so that the pipeline of the injection pump and the switching valve is filled with the isolation liquid.

[0023] In the system provided by one embodiment, the subsystem also includes a stirring device for stirring the liquid in the culture container; the stirring device includes a motor, a rotating member and a driven member; the rotating member and the driven member are both magnetic devices, at least one of which is a magnet; the driven member is used to be placed in the culture container, and the motor is used to drive the rotating member to rotate, and when the rotating member rotates, the driven member is driven to rotate through the magnetic field.

[0024] In the system provided by an embodiment, the subsystem further includes:

[0025] a detection device for detecting the absorbance of the liquid in the culture container; and / or,

[0026] A constant temperature device is used to keep the temperature of the culture container constant.

[0027] In the system provided by one embodiment, the culture medium providing device comprises:

[0028] A culture medium container, used for containing culture medium;

[0029] Peristaltic pumps;

[0030] The culture medium container is connected to the culture container via the peristaltic pump, and the peristaltic pump is used to extract the culture medium from the culture medium container and output the extracted culture medium to the culture container.

[0031] The system provided in one embodiment further includes:

[0032] A control terminal, used to receive control parameters of the subsystem input by a user, and control the operation of the subsystem according to the input control parameters; and / or,

[0033] The second container is used to hold the inducer; the second container is connected to a channel opening of the switching valve.

[0034] The microbial culture system according to the above embodiment includes at least one subsystem. The subsystem includes: a culture container, a sampling container, a first container, a second container, a third container, a liquid storage ring, an injection pump, a switching valve with a common channel port and multiple channel ports, a culture medium supply device and a liquid level maintaining device. The injection pump cooperates with the switching valve to realize the suction of liquids in each container; the culture medium supply device can provide culture medium to the culture container; the liquid level maintaining device can suck away excess liquid when the liquid level in the culture container exceeds a preset height so that the liquid level in the culture container does not exceed the preset height. It can be seen that suction of liquid, supply of culture medium and maintenance of the volume of microbial solution can be realized by the microbial culture system without the need for manual operation by the user, thereby improving efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 A structural block diagram of an embodiment of a microorganism cultivation system provided by the utility model;

[0036] Figure 2 A schematic diagram of a liquid path of a subsystem 1 in a microorganism culture system provided by the utility model;

[0037] Figure 3 A schematic diagram of an embodiment of a liquid level maintaining device in the microorganism culture system provided by the utility model;

[0038] Figure 4A schematic diagram of an embodiment of a stirring device in the microorganism culture system provided by the utility model;

[0039] Figure 5 This is a flow chart of an embodiment of a control terminal control subsystem in the microorganism cultivation system provided by the utility model;

[0040] Figure 6 A schematic diagram of an embodiment of a detection device in the microorganism culture system provided by the utility model;

[0041] Figure 7 This is a graph showing the growth rate changing over time in the microbial culture system provided by the utility model. DETAILED DESCRIPTION

[0042] The present invention is further described in detail below by specific embodiments in conjunction with the accompanying drawings. Wherein similar elements in different embodiments adopt associated similar element numbers. In the following embodiments, many detailed descriptions are intended to enable the present application to be better understood. However, those skilled in the art can easily recognize that some of the features can be omitted in different situations, or can be replaced by other elements, materials, and methods. In some cases, some operations related to the present application are not shown or described in the specification, in order to avoid the core part of the present application being overwhelmed by too much description, and for those skilled in the art, it is not necessary to describe these related operations in detail, and they can fully understand the related operations based on the description in the specification and the general technical knowledge in the art.

[0043] In addition, the features, operations or characteristics described in the specification can be combined in any appropriate manner to form various implementations. At the same time, the steps or actions in the method description can also be interchanged or adjusted in a manner that is obvious to those skilled in the art. Therefore, the various sequences in the specification and the drawings are only for the purpose of clearly describing a certain embodiment and are not meant to be a required sequence, unless otherwise specified that a certain sequence must be followed.

[0044] The serial numbers of the components in this document, such as "first", "second", etc., are only used to distinguish the objects described and do not have any order or technical meaning. The "connection" and "coupling" mentioned in this application, unless otherwise specified, include direct and indirect connections (couplings).

[0045] The microorganism culture system provided by the utility model is as follows: Figure 1 As shown, at least one subsystem 20 is included.

[0046] Please also read Figure 2The subsystem 20 includes: a culture container 24, a sampling container 28, a first container 31, a second container 32, a third container 34, a liquid storage ring 29, an injection pump 30, a switching valve 33, a culture medium supply device 25, a detection device 22 and a liquid level maintaining device 39.

[0047] The culture container 24 is used to provide a place for the evolution of microorganisms, that is, it can contain liquid containing microorganisms and culture medium, and it can be various types of containers, such as culture bottles and the like.

[0048] The sampling container 28 is used to hold the liquid (sample solution) extracted from the culture container 24 by the injection pump 30. The number of the sampling container 28 can be one or more (two or more), etc. In this embodiment, multiple sampling containers 28 are provided so that sample solutions obtained at different times can be held by multiple sampling containers 28. Similarly, the sampling container 28 can hold the sample solution, and it can be various types of containers, such as sampling bottles, etc.

[0049] The first container 31 is used to hold a growth inhibitor. The growth inhibitor is used to inhibit the growth of microorganisms. The microorganisms may be bacteria, and the corresponding growth inhibitor may be an antibiotic. Accordingly, the evolution experiment of the microorganism may be: allowing bacteria (such as Escherichia coli) to grow and reproduce in an antibiotic environment, and producing drug resistance after multiple generations of cultivation, so that the bacteria evolve drug resistance genes.

[0050] The second container 32 is used to hold an inducer, which can be used to promote the microorganisms to produce corresponding proteins.

[0051] The third container 34 is used to hold a cleaning liquid. The cleaning liquid is used to clean various containers and pipelines in the subsystem 20, and the cleaning liquid may be a sodium hydroxide solution.

[0052] The liquid storage ring 29 is used to store the liquid sucked by the injection pump 30. It can be a long tube, usually surrounded in a ring shape or a disk shape to save space.

[0053] The syringe pump 30 is used to provide power for the flow of liquid in the liquid circuit of the subsystem.

[0054] The switching valve 33 has a common channel port and a plurality of channel ports. Figure 2 As shown, each channel port is distributed around the common channel port. The switching valve 33 is used to selectively connect one of the multiple channel ports to the common channel port. In other words, the switching valve 33 switches the channel port so that the switched channel port is connected to the common channel port, while the other channel ports are not connected to the common channel port.

[0055] The syringe pump 30 is connected to the common channel port of the switching valve 33 through the liquid storage ring 29, while the culture container 24, the sampling container 28, the first container 31, the second container 32 and the third container 34 are respectively connected to different channel ports of the switching valve 33. It can be seen that the switching valve 33 can realize that the syringe pump 30 is connected to different containers at different times by switching, so as to realize liquid aspiration and liquid injection.

[0056] The culture medium supply device 25 is used to supply culture medium to the culture container 24 .

[0057] The liquid level maintaining device 39 is used to absorb excess liquid when the liquid level in the culture container 24 exceeds a preset height so that the liquid level in the culture container 24 does not exceed the preset height.

[0058] It can be seen that aspirating liquid, providing culture medium and maintaining the volume of microbial solution can be achieved by the microbial culture system without the need for manual operation by the user, thereby improving efficiency.

[0059] In one embodiment, the microorganism cultivation system may further include a control terminal 10. The control terminal 10 is connected to each subsystem in communication. The control terminal 10 is used to control each subsystem 20. In this embodiment, the control terminal 10 has a human-computer interaction device for human-computer interaction. The human-computer interaction device includes a display and an input device, and the input device is used to receive the user's input operation, which may include a mouse, a keyboard, a touch screen, etc. The user can control each subsystem 20 through the control terminal 10. The control terminal 10 can be any device with data processing function and human-computer interaction function, such as various types of computers.

[0060] The subsystem 20 is used to cultivate microorganisms under the control of the control terminal 10. That is, the hardware composition of the microorganism cultivation system is mainly the subsystem 20.

[0061] The control terminal 10 is electrically connected to the injection pump 30, so that the injection pump 30 can be controlled. In this embodiment, the control terminal 10 controls multiple subsystems 20. Considering that various pumps and valves need to be driven for control, as well as the need to control multiple subsystems 20, each subsystem also includes a controller (slave). The controller is used to communicate with the control terminal 10, receive control instructions from the control terminal 10, and control the subsystem accordingly. That is, the control terminal 10 controls the pumps, valves and other devices in the subsystem 20 through the controller.

[0062] The control terminal 10 is electrically connected to the switching valve 33 , so that the switching valve 33 can be controlled to switch. Similarly, in this embodiment, the control terminal 10 controls the switching valve 33 through a controller.

[0063] In this embodiment, the culture medium providing device 25 includes: a culture medium container 251 and a peristaltic pump 252 .

[0064] The culture medium container 251 is used to hold the culture medium and is connected to the culture container 24 via a peristaltic pump 252 .

[0065] The peristaltic pump 252 is used to extract the culture medium from the culture medium container 251 and output the extracted culture medium to the culture container 24. The control terminal 10 is electrically connected to the peristaltic pump 252, so as to control the peristaltic pump 252. Similarly, in this embodiment, the control terminal 10 controls the peristaltic pump 252 through the controller.

[0066] In order to balance the pressure, the culture medium container 251 is also connected to the atmosphere through the filter component 38. The filter component 38 is used to filter the air, and in this embodiment, a filter membrane is used.

[0067] The detection device 22 is used to detect the concentration of the liquid in the culture container 24, and the concentration reflects the number of microorganisms, and the two are proportional. The control terminal 10 is electrically connected to the detection device 22 and receives the concentration detected by the detection device 22. In this embodiment, the control terminal 10 receives the concentration detected by the detection device 22 through the controller.

[0068] The control terminal 10 is used to receive the control parameters of the subsystem input by the user, and control the operation of the subsystem according to the input control parameters. When there are multiple subsystems, the user can input control parameters for each subsystem respectively, so that different subsystems can work under different control parameters and conduct different experiments, thereby improving the efficiency of the experiment. From the above content, it can be seen that in the microbial culture system of the utility model, the injection pump 30 can selectively absorb the solution, growth inhibitor, inducer and cleaning agent in the culture container 24, and automatically carry out microbial culture under the control of the control terminal 10, such as automatically carrying out continuous evolution experiments, with a high degree of automation, which improves the experimental efficiency.

[0069] The control parameters may include a set concentration, and the control terminal 10 may control the peristaltic pump 252 through a preset PID algorithm so that the concentration detected by the detection device is maintained at the set concentration.

[0070] The control terminal 10 is electrically connected to the liquid level maintaining device 39 and can control the liquid level maintaining device 39. The liquid level maintaining device 39 is also used to suck external air into the culture container 24 when the liquid level in the culture container 24 does not exceed a preset height. In this way, the volume of the solution in the culture container 24 can be kept stable and the oxygen content in the culture container 24 can be guaranteed.

[0071] Please also read Figure 3 The liquid level maintaining device 39 includes a vacuum pump 35. The suction end of the vacuum pump 35 is connected to the culture container 24 through a first liquid pipette 391. The first liquid pipette 391 extends into the culture container 24, and the height of the bottom end of the first liquid pipette 391 is a preset height. This method is simple and reliable to maintain the stability of the volume of the microbial solution.

[0072] The culture container 24 can be connected to the atmosphere through a filter element 38, such as Figure 3 As shown, when the vacuum pump 35 is working, on the one hand, it can absorb excess microbial solution, and on the other hand, it can suck external air into the culture container 24 to maintain the oxygen content, thus killing two birds with one stone.

[0073] Considering that the air filtered by the membrane may contain pollutants such as dust and microorganisms, which may affect the purity of the experiment, and directly extracting air from the external environment may cause fluctuations in the humidity and temperature of the microbial solution (culture solution), affecting the growth environment of the microorganisms. Therefore, this embodiment further filters the air through water, and allows the air entering the culture container 24 to maintain a certain temperature and humidity. Figure 2 As shown, in this embodiment, the liquid level maintaining device 39 further comprises a water storage container 26 for storing water. The "water" here can be ultrapure water.

[0074] The water storage container 26 is connected to the culture container 24. As shown in the figure, most of the connections in this article are through pipeline connections (liquid circuit connections). An air inlet pipe 261 is provided in the water storage container 26, one end of the air inlet pipe 261 is connected to the atmosphere through a filter component 38, and the other end of the air inlet pipe 261 extends below the water surface. In this way, driven by the vacuum pump 35, the external air passes through the filter membrane 38, then passes through ultrapure water, and then enters the culture container 24. Ultrapure water can further remove microorganisms and particulate matter that may remain in the air, thereby improving the cleanliness of the air entering the culture container 24. Since the air passes through ultrapure water first, the fluctuations directly introduced from the external environment can be reduced, and the stability of the culture solution can be maintained. The intervention of ultrapure water can reduce the potential impact of harmful substances in the air on microorganisms and reduce stress reactions.

[0075] The subsystem may further include a waste liquid container 36, which is connected to a channel port of the switching valve 33 and is also connected to the discharge end of the vacuum pump 35. The switching valve 33 can be switched to allow the syringe pump 30 to communicate with the waste liquid container 36, thereby discharging the waste liquid into the waste liquid container 36.

[0076] The liquid level maintaining device 39 also includes a three-way valve 37. The suction end of the vacuum pump 35 is connected to the outlet end I of the three-way valve 37, the first inlet end II of the three-way valve 37 is connected to the culture container 24 through a first liquid suction pipe 391, and the second inlet end III of the three-way valve 37 is connected to the culture container 24 through a second liquid suction pipe 392. The second liquid suction pipe 392 extends into the bottom of the culture container 24. The three-way valve 37 selectively connects one of the first inlet end II and the second inlet end III to the outlet end I. During the experiment, the three-way valve 37 connects the first inlet end II with the outlet end I, so that the vacuum pump 35 maintains the liquid level and oxygen content of the microbial solution. After the experiment, the three-way valve 37 connects the second inlet end III with the outlet end I, and the vacuum pump 35 pumps the microbial solution into the waste liquid container 36. The control terminal 10 is electrically connected to the vacuum pump 35 and the three-way valve 37 to control the vacuum pump 35 and the three-way valve 37, for example, the vacuum pump 35 and the three-way valve 37 are controlled by a controller. The three-way valve 37 may be a solenoid valve, which is convenient for the control terminal 10 to control.

[0077] The control terminal 10 controls the speed of the vacuum pump 35 to adjust the oxygen supply of the microorganisms. Specifically, the control parameter can be the oxygen supply. The control terminal 10 matches the speed corresponding to the oxygen supply input by the user according to the preset correspondence between the oxygen supply and the speed, thereby controlling the vacuum pump 35 to work at the matched speed, so that the oxygen supply of the microorganisms meets the user's expectations.

[0078] The subsystem also includes a fourth container 31 for holding an isolation liquid, and the isolation liquid 31 is used to isolate the liquid and air in other containers. Isolating the liquid in other containers can prevent the injection pump 30 from being contaminated, and isolating the air can prevent the accuracy of the injection pump 30 in conveying liquid from being affected by the air, and the accuracy is high. In this embodiment, the isolation liquid 31 is oil. The fourth container 31 is connected to a channel port of the switching valve 33, and the switching valve 33 can be switched to connect the injection pump 30 with the fourth container 31, so as to extract the isolation liquid 31. The injection pump 30 fills the pipeline of the injection pump 30 and the switching valve 33 with the isolation liquid by sucking the isolation liquid in the fourth container 31, so that the injection pump 30 can suck the liquid in other containers through the cooperation of the switching valve 33.

[0079] The subsystem also includes a stirring device 21 for stirring the liquid (microorganism solution) in the culture container 24. In existing stirring devices, the rotating part is usually inside the bottle, and the motor and other electrical appliances are outside the bottle. This requires the drive shaft and other components to pass through the bottle. Considering the sealing requirements, the bottle and the stirring device can only be made into a whole, which is very inconvenient to clean and prone to sealing problems. This embodiment does not use a mechanical transmission mechanism, so the culture container 24 and the stirring device 21 are separate and independent of each other, which makes disassembly and cleaning very convenient, and there is no need to consider sealing issues. Specifically, if Figure 4As shown, the stirring device 21 includes a motor 211, a rotating member 212 and a driven member 213. The rotating member 212 and the driven member 213 are both magnetic devices, at least one of which is a magnet. The magnet can be a permanent magnet or an electromagnet, as long as it can generate a magnetic field, so that the two can affect each other through the magnetic field. The driven member 213 is used to be placed in the culture container 24, and usually its density is greater than that of water, and it sinks to the bottom of the microbial solution. The motor 211 is used to drive the rotating member 212 to rotate, and when the rotating member 212 rotates, the driven member 213 is driven to rotate through the magnetic field. In this embodiment, the magnetic poles at both ends of the rotating member 212 are opposite.

[0080] The control parameter may include a stirring speed, and the control terminal 10 controls the motor 211 according to the stirring speed, so that the motor 211 works at the stirring speed. In this way, the user can flexibly set the stirring speed.

[0081] The subsystem also includes a thermostat 23 for maintaining a constant temperature of the culture container. The control parameters may include a growth temperature. In this way, after the user inputs the growth temperature, the control terminal 10 controls the thermostat 23 to heat the culture container 24 according to the growth temperature input by the user, so that the temperature of the culture container 24 is maintained at the growth temperature, which is convenient for the microorganisms to reproduce at a constant temperature. Different microorganisms may have different optimal growth temperatures. Therefore, the control terminal 10 may pre-store the optimal growth temperatures of different microorganisms. The control terminal 10 is also used to receive the type of microorganism input by the user, find the optimal growth temperature corresponding to the microorganism type in the pre-stored optimal growth temperatures of different microorganisms, and display the optimal growth temperature corresponding to the microorganism type, so that the user can set the growth temperature. Provide a reference. The control terminal 10 controls the thermostat 23 to heat the culture container 24. Specifically, the thermostat 23 can be controlled to heat the culture container 24 based on a preset PID algorithm, and the temperature can be accurately controlled to remain at the growth temperature (with an error of about 0.3 degrees Celsius).

[0082] The control parameters may also include a first concentration threshold. The control terminal 10 controls the subsystem to work according to the first concentration threshold input by the user, such as Figure 5 As shown, the following steps are included:

[0083] Step 1: The control terminal 10 determines whether the concentration detected by the detection device 22 is greater than the first concentration threshold. If so, step 2 is executed. If the concentration detected by the detection device 22 is not greater than the first concentration threshold, no action is taken and the microorganisms are allowed to grow on their own.

[0084] Step 2: When the concentration detected by the detection device 22 is higher than the first concentration threshold, the control terminal 10 controls the injection pump 30 and the switching valve 33 to inject the growth inhibitor into the culture container 24. Specifically, the control terminal 10 controls the switching valve 33 to connect the channel port connected to the first container 31 with the common channel port, so that the injection pump 30 is connected to the first container 31, thereby controlling the injection pump 30 to absorb the growth inhibitor from the first container 31 and store it in the liquid storage ring 29, and then controls the switching valve 33 to connect the channel port connected to the culture container 24 with the common channel port, so that the injection pump 30 is connected to the culture container 24, and controls the injection pump 30 to inject the growth inhibitor in the liquid storage ring 29 into the culture container 24. The liquid in other containers is injected into the culture container 24, and the liquid in the culture container 24 is injected into the sampling container 28, and the operation process of the injection pump 30 and the switching valve 33 is similar to this. Taking the evolution experiment as an example, the user only needs to set the first concentration threshold of the microorganism. After the liquid concentration in the culture container 24 is higher than the first concentration threshold, antibiotics will be automatically added without the need for manual operation by the user. After multiple antibiotic screenings, microorganisms that have evolved antibiotic-resistant genes can multiply in large numbers, thereby obtaining the desired samples.

[0085] When the concentration detected by the detection device 22 is higher than the first concentration threshold, the control terminal 10 also controls the culture medium providing device 25 to provide culture medium to the culture container 24, specifically, controls the peristaltic pump 252 to work, and the peristaltic pump 252 extracts the culture medium from the culture medium container 251 and outputs the extracted culture medium to the culture container 24. Injecting the culture medium into the culture container 24 can not only dilute the liquid in the culture container 24, but also provide nutrition to the microorganisms, so that the microorganisms with drug resistance can reproduce as soon as possible after being screened by antibiotics. After adding the culture medium, as described above, the excess bacterial liquid in the culture container 24 will be discharged into the waste liquid bottle 36.

[0086] There are many ways to detect the concentration of the liquid in the culture container 24 by the detection device 22. In this embodiment, the detection device 22 specifically detects the absorbance (OD value) of the liquid in the culture container 24, and uses the absorbance to reflect the concentration. When the concentration of the bacterial liquid increases, the light intensity passing through the culture container 24 will become weaker, and the OD value will increase. The detection device 22 can accurately measure the OD value of the bacterial liquid through a calibration experiment, and can also accurately correspond to the concentration.

[0087] like Figure 6 As shown, the detection device 22 of this embodiment includes a laser light source 221, a beam splitter 222, a first light intensity sensor 223, a second light intensity sensor 224 and a processor. The three dotted arrows in the figure indicate the installation positions of the beam splitter 222, the first light intensity sensor 223 and the second light intensity sensor 224 respectively.

[0088] The laser light source 221 is used to emit a laser beam.

[0089] The beam splitter 222 is disposed on the optical path of the laser beam, and is used to split the laser beam into a first sub-laser beam and a second sub-laser beam with different directions, wherein the first sub-laser beam is directed toward the culture container 24 .

[0090] The first light intensity sensor 223 is used to detect the light intensity of the first sub-laser beam after passing through the culture container 24.

[0091] The second light intensity sensor 224 is used to detect the light intensity of the second laser sub-beam. The light intensity of the second laser sub-beam and the first laser sub-beam respectively represent the light intensity of the laser from the laser light source 221 and the light intensity of the laser after passing through the microbial solution, so the two light intensities can be used to calculate the absorbance.

[0092] The processor is used to calculate the absorbance according to the light intensity detected by the first light intensity sensor 223 and the second light intensity sensor 224 .

[0093] Accordingly, in the above content, the concentration detected by the detection device 22 is specifically the detected absorbance, and the first concentration threshold is specifically the first absorbance threshold. Figure 5 The concentration in the process can be concretized as absorbance, and the first concentration threshold can be concretized as the first absorbance threshold.

[0094] The control terminal 10 is also used to calculate the growth rate of the microorganism based on a plurality of different concentrations and their corresponding times. Specifically, the control parameter may also include a second concentration threshold (such as a second absorbance threshold), and the second concentration threshold is less than the first concentration threshold. The control terminal 10 records the corresponding times t1 and t2 when the concentration detected by the detection device 22 reaches the first concentration threshold M1 and the second concentration threshold M2, respectively, and calculates the growth rate based on M1, M2, t1 and t2. In this embodiment, the growth rate is characterized by the doubling time Tau, and the growth rate Tau is calculated specifically according to the following formula:

[0095] Tau=(t1-t2) / [(l gM1-l gM2) / l g2];

[0096] If the concentration is expressed in terms of absorbance, M1 is the first absorbance threshold, and M2 is the second absorbance threshold. Both the concentration threshold and the absorbance threshold representing the concentration can be set by the user, so that the user can set it according to different microorganisms and different experimental purposes, which is very convenient. In this embodiment, the first absorbance threshold is 0.7 and the second absorbance threshold is 0.5.

[0097] Each time antibiotics are added, microorganisms are screened, and the growth rate is calculated during the recovery of the microbial population. The control terminal 10 can generate a trend graph of growth rate changes over time based on the calculated multiple growth rates and their corresponding times, that is, generate a growth rate change curve, such as Figure 7 As shown, the trend graph is displayed. Figure 7 It can be clearly seen that the growth rate of the microorganisms is increasing. The control terminal 10 can also display the change curves of the growth rates of each subsystem in the same trend graph, so that the differences in the growth rates of each subsystem can be compared at the same time coordinate. In this case, it is suitable for the user to set the same variable for different subsystems, so as to understand the image of the variable on the growth rate of the microorganism by comparing the growth rates of each subsystem.

[0098] When the growth rate reaches the preset threshold, the control terminal 10 controls the subsystem to end the experiment, for example, the control subsystem performs sampling and then cleaning. Specifically, the control terminal 10 controls the switching valve 33 to connect the channel port connected to the culture container 24 with the common channel port, so that the injection pump 30 is connected to the culture container 24, and the injection pump 30 is controlled to absorb the liquid in the culture container 24 and store it in the liquid storage ring 29. Then, the switching valve 33 is controlled to connect the channel port connected to the sampling container 28 with the common channel port, so that the injection pump 30 is connected to the sampling container 28, and the injection pump 30 is controlled to inject the absorbed liquid (liquid stored in the liquid storage ring 29) into the sampling container 28, thereby completing the sampling. It can be seen that the user only needs to set the control parameters, and the entire system can automatically run based on the control parameters until the experiment is completed, which is very efficient.

[0099] The subsystem is cleaned after sampling, specifically, the control terminal 10 controls the three-way valve 37 to connect its outlet port I with the second inlet port III, and controls the vacuum pump 35 to discharge the remaining liquid in the culture container 24 into the waste liquid container 36. Then, the control terminal 10 controls the switching valve 33 to connect the channel port connected to the third container 34 with the common channel port, so that the injection pump 30 is connected to the third container 34, and the injection pump 30 is controlled to absorb the cleaning liquid in the third container 34 and store it in the liquid storage ring 29. Then, the switching valve 33 is controlled to connect the channel port connected to the culture container 24 with the common channel port, so that the injection pump 30 is connected to the culture container 24, and the injection pump 30 is controlled to inject the absorbed cleaning liquid into the culture container 24 for cleaning. After cleaning, the vacuum pump 35 is controlled to work to discharge the liquid in the culture container 24 into the waste liquid container 36.

[0100] The user can also input the sampling conditions into the control terminal 10, and the control terminal 10 performs sampling according to the sampling conditions. For example, the sampling in the aforementioned content corresponds to the sampling condition that the growth rate reaches a preset threshold. If the sampling condition input by the user is: sampling at the sampling concentration input by the user, the control terminal 10 controls the subsystem to sample when the concentration detected by the detection device reaches the sampling concentration. The sampling process has been described in the aforementioned content and will not be repeated here. If the sampling condition input by the user is: sampling at the sampling time input by the user, the control terminal 10 uses the sampling time as the time interval to periodically control the subsystem to sample.

[0101] Directed evolution is a powerful method in molecular bioengineering. It is used to modify the function, structure and stability of proteins in order to enhance or expand protein activity. It has extremely important application value in academic research and biotechnology. Traditional directed evolution relies heavily on random mutation of genes in vitro, which requires the use of error-prone PCR or random oligonucleotide libraries. This process is often labor-intensive, expensive, time-consuming and labor-intensive.

[0102] Ideal continuous directed evolution is a high-frequency mutation within the target gene range, which can simultaneously achieve gene diversification and corresponding screening in cells. This technology will significantly enhance the in-depth study of the evolution of biological macromolecules. There are already a variety of in vivo targeted high-frequency mutation methods based on different principles in the prior art. These methods achieve the continuous introduction of mutations in the specified region of the gene, thus realizing the ideal strategy of directed and continuous mutagenesis. However, the experimental system used to implement these methods requires researchers to manually replace the culture medium at a fixed time every day to achieve continuous growth and rapid division of microorganisms in the logarithmic phase. The experimental cycle is long, often taking several days to weeks, which is a large workload and takes up a lot of time of scientific researchers.

[0103] As can be seen from the above contents, the microorganism cultivation system provided by the present invention can improve the automation level of microorganism cultivation, especially microorganism evolution, thereby improving efficiency.

[0104] The above specific examples are used to illustrate the present invention, which are only used to help understand the present invention and are not intended to limit the present invention. For those skilled in the art of the present invention, some simple deductions, deformations or substitutions can be made based on the idea of ​​the present invention.

Claims

1. A microorganism cultivation system, characterized in that: It comprises at least one subsystem; the subsystem comprises: Culture vessels, used to provide a place for microorganisms to evolve; A sampling container, used for holding the sample solution drawn from the culture container by the syringe pump; A first container for containing a growth inhibitor; A third container is used to hold a cleaning solution; Liquid storage ring; Syringe pump; A switching valve having a common channel port and a plurality of channel ports, wherein the injection pump is connected to the common channel port of the switching valve through the liquid storage ring, and the culture container, the sampling container, the first container and the third container are respectively connected to different channel ports of the switching valve; the switching valve is used to selectively connect one of the plurality of channel ports to the common channel port; A culture medium supply device, used for supplying culture medium to the culture container; The liquid level maintaining device is used to suck away excess liquid when the liquid level in the culture container exceeds a preset height so that the liquid level in the culture container does not exceed the preset height.

2. The system according to claim 1, characterized in that The liquid level maintaining device is also used to suck external air into the culture container when the liquid level in the culture container does not exceed a preset height.

3. The system according to claim 2, characterized in that The liquid level maintaining device comprises: A vacuum pump; the suction end of the vacuum pump is connected to the culture container through a first pipette; the first pipette extends into the culture container, and the height of the bottom end of the first pipette is the preset height.

4. The system according to claim 3, characterized in that The liquid level maintaining device also includes: A water storage container is used for storing water; the water storage container is connected to the culture container; an air inlet pipe is arranged in the water storage container, one end of the air inlet pipe is connected to the atmosphere through a filter component, and the other end of the air inlet pipe extends below the water surface.

5. The system according to claim 3 or 4, characterized in that The subsystem further comprises a waste liquid container, which is connected to a channel port of the switching valve and is also connected to a discharge end of the vacuum pump; The liquid level maintaining device also includes a three-way valve; the suction end of the vacuum pump is connected to the outlet end of the three-way valve, the first inlet end of the three-way valve is connected to the culture container through a first pipette, the second inlet end of the three-way valve is connected to the culture container through a second pipette, and the second pipette extends into the bottom of the culture container; the three-way valve selectively connects one of the first inlet end and the second inlet end to the outlet end.

6. The system according to claim 1, wherein: The subsystem also includes a fourth container for holding an isolation liquid, and the isolation liquid is used to isolate the liquid and air in other containers; the fourth container is connected to a channel opening of the switching valve; the injection pump sucks the isolation liquid in the fourth container so that the pipelines of the injection pump and the switching valve are filled with the isolation liquid.

7. The system according to claim 1, characterized in that The subsystem also includes a stirring device for stirring the liquid in the culture container; the stirring device includes a motor, a rotating member and a driven member; the rotating member and the driven member are both magnetic devices, at least one of which is a magnet; the driven member is used to be placed in the culture container, and the motor is used to drive the rotating member to rotate, and when the rotating member rotates, the driven member is driven to rotate through the magnetic field.

8. The system of claim 1, wherein: The subsystem also includes: a detection device for detecting the absorbance of the liquid in the culture container; and / or, A constant temperature device is used to keep the temperature of the culture container constant.

9. The system according to claim 1, characterized in that The culture medium providing device comprises: A culture medium container, used for containing culture medium; Peristaltic pumps; The culture medium container is connected to the culture container via the peristaltic pump, and the peristaltic pump is used to extract the culture medium from the culture medium container and output the extracted culture medium to the culture container.

10. The system according to claim 1, wherein: Also includes: A control terminal, used to receive control parameters of the subsystem input by a user, and control the operation of the subsystem according to the input control parameters; and / or, The second container is used to hold the inducer; the second container is connected to a channel opening of the switching valve.

Images