A swing type visual high-pressure microbial batch culture device and method

Abstract

The application provides a swing type visual high-pressure microbial batch culture device, which comprises a culture system and a swing system, wherein the culture system comprises a culture cabin, visual observation windows are arranged on two sides of the culture cabin, the swing system comprises a reduction motor, a main body support and a digital time control instrument, the main body support is used for fixing the culture cabin, the reduction motor drives the main body support to rotate, and the culture cabin swings uniformly, the culture cabin is driven by the swing system to swing by ±90 degrees or ±180 degrees, problems that organisms are prone to sedimentation, gathering and local nutrient exhaustion during long-time culture can be avoided, and the visual observation windows made of sapphire can be used for observing the culture condition in the culture cabin in real time.

Description

Technical Field

[0001] This invention relates to the field of microbial culture technology, and in particular to a swing-type visual high-pressure microbial batch culture device and method. Background Technology

[0002] Deep-sea habitats are among the most biodiverse ecosystems on Earth, and deep-sea microorganisms play an indispensable role in them. They participate in energy and material cycles within deep-sea ecosystems. Current research on deep-sea microorganisms focuses on biodiversity, biotechnological potential, ecosystem function, and adaptability to extreme environments. A deeper understanding of the life processes of these microorganisms in extreme environments and their contributions to Earth's ecosystems is of great significance.

[0003] Multi-omics methods can be used to analyze thousands of quantitative measurement data to gain insights at the molecular level. However, before omics analysis, the target microorganisms need to be cultured under deep-sea simulated conditions. Existing technologies for microbial culture include single-pot culture and batch culture, but most of them use static culture. During long-term culture, there are problems such as easy sedimentation, aggregation, and local nutrient depletion of organisms. At the same time, it is impossible to observe the culture status of microorganisms during the culture process. Summary of the Invention

[0004] To address the aforementioned problems, this invention proposes a swing-type visual high-pressure microbial batch culture device and method.

[0005] This invention is achieved through the following technical solution: This invention proposes a swing-type visual high-pressure microbial batch culture device, comprising a culture system and a swing system, wherein: The culture system includes a culture chamber with visual observation windows on both sides. The swaying system includes a geared motor, a main support frame, and a digital display time control instrument. The main support frame is used to fix the culture chamber, and the geared motor drives the main support frame to rotate, causing the culture chamber to sway at a uniform speed.

[0006] Furthermore, the culture chamber includes a high-pressure chamber body, an upper flange, and a lower flange. The upper flange and the lower flange are fixed to the top and bottom of the high-pressure chamber body, respectively, and form a culture cavity inside the high-pressure chamber body. A test tube rack is provided inside the culture cavity, and culture test tubes are provided on the test tube rack.

[0007] Furthermore, both the upper flange and the lower flange are provided with multiple connection ports, which communicate with the culture chamber. The upper flange is provided with a sensor mounting slot, which is a blind slot extending along the axial direction of the upper flange.

[0008] Furthermore, it also includes a pressurization system, which includes a constant pressure pump. One end of the constant pressure pump is connected to an explosion-proof valve, and the explosion-proof valve is connected to the connection port on the lower flange to the inside of the culture chamber.

[0009] Furthermore, it also includes a monitoring system, which includes a temperature sensor and a pressure sensor. The temperature sensor is installed inside the sensor mounting groove of the upper flange, and the pressure sensor is installed inside the culture chamber through the connection port of the upper flange. The temperature sensor and the pressure sensor are connected to a digital display time control instrument. A drain valve is connected to one connection port of the lower flange, and a vent valve is connected to one connection port of the upper flange.

[0010] Furthermore, it also includes a temperature control system, which includes a water jacket located on the outside of the culture chamber and connected to a constant temperature water bath system to form a circulation loop.

[0011] Furthermore, the water jacket is a spiral circulation structure that surrounds the outside of the culture chamber, and the outermost part of the water jacket is covered with a heat insulation cover.

[0012] Furthermore, it also includes a sampling device, which is equipped with a fixing clamp and a lead screw. The fixing clamp is controlled by a pneumatic clamping knob to fix the culture tube, and the lead screw pushes the pistons on both sides of the test tube upward under the drive of the control panel, thereby opening the test tube.

[0013] Furthermore, a swing-type visualized high-pressure microbial batch culture method includes the following steps: S1. Turn on the constant temperature water bath system, set the experimental temperature, and maintain constant temperature control in the culture chamber; S2. Sterilize the culture tubes used in the experiment, fill the microbial samples in an anaerobic glove box, and place the culture tubes on the test tube rack; S3. Open the upper flange and install the test tube rack and cushioning cotton for buffering, then re-secure the upper flange; S4. Connect the constant pressure pump, drain valve, pressure sensor, temperature sensor and explosion-proof valve, and seal the culture chamber; S5. Open the vent valve and constant pressure pump on the upper flange, set the pressure value and run the constant pressure pump. When water flows out in streams at the outlet of the vent valve, close the vent valve and apply a constant pressure to the culture chamber to the set pressure. S6. Perform microbial culture and thoroughly mix the sample solution using a shaking device; S7. After the culture experiment is completed, set the constant pressure pump to 0.1MPa and start it. Open the drain valve and the vent valve, and then open the upper flange to take out the test tube rack. S8. Place the culture tube into the sampling device and clamp it, then push the pistons on both sides of the culture tube to move and pour out the sample.

[0014] The beneficial effects of this invention are: (1) The swing-type visual high-pressure microbial batch culture proposed in this invention uses a swing system to drive the culture chamber to swing at ±90 degrees or ±180 degrees, which can avoid the problems of easy sedimentation, aggregation and local nutrient depletion of organisms during long-term culture. At the same time, the sapphire material visualization observation window can be used to observe the culture situation inside the culture chamber in real time.

[0015] (2) The swing-type visual high-pressure microbial batch culture device proposed in this invention uses a water jacket and a constant temperature water bath system to form a circulation loop, which can achieve constant temperature control of the culture chamber and prevent temperature changes from affecting the experimental results.

[0016] (3) The main body of the swing-type visual high-pressure microbial batch culture device proposed in this invention can be moved by rollers. Compared with the existing large-volume high-pressure biological culture device, it is not limited by weight and size, and can be moved to different locations, thus having greater flexibility. Attached Figure Description

[0017] Figure 1 This is an overall structural diagram of the swing-type visual high-pressure microbial batch culture device of the present invention; Figure 2 This is a structural diagram of the sampling device of the swing-type visual high-pressure microbial batch culture device of the present invention; In the diagram: 1. Digital display time control instrument; 2. Constant pressure pump; 3. Gear motor; 4. Main support; 5. Visual observation window; 6. Water jacket; 7. High pressure chamber body; 8. Upper flange; 9. Lower flange; 10. Pressure sensor; 11. Temperature sensor; 12. Drain valve; 13. Connection port; 14. Fixing clamp; 15. Screw; 16. Constant temperature water bath system; 17. Culture tube; 18. Sensor mounting slot; 19. Venting valve. The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0018] To more clearly and completely illustrate the technical solution of the present invention, the present invention will be further described below with reference to the accompanying drawings.

[0019] Please refer to Figure 1-2 This invention proposes a swing-type visual high-pressure microbial batch culture device, comprising a culture system and a swing system, wherein: The culture system includes a culture chamber with visual observation windows 5 on both sides. The swaying system includes a geared motor 3, a main support 4, and a digital display time control instrument 1. The main support 4 is used to fix the culture chamber, and the geared motor 3 drives the main support 4 to rotate, so that the culture chamber sways at a uniform speed.

[0020] In a specific embodiment, symmetrically arranged visual observation windows 5 are provided on both sides of the culture chamber. The visual observation windows 5 are made of sapphire material. A swing system is used to drive the culture chamber to swing, so that the liquid inside the culture chamber is fully mixed during the culture process. The swing system includes a geared motor 3, a main support 4, and a digital display time control instrument 1. The culture chamber is set on the main support 4. The geared motor 3 drives the main support 4 to rotate ±90 degrees or ±180 degrees. The geared motor 3 is controlled by the digital display time control instrument 1. This invention uses a swing system to drive the culture chamber to rotate ±90 degrees or ±180 degrees, which can avoid the problems of easy sedimentation, aggregation, and local nutrient depletion of organisms during long-term culture. At the same time, the visual observation windows 5 made of sapphire material can be used to observe the culture status inside the culture chamber in real time.

[0021] Furthermore, the culture chamber includes a high-pressure chamber body 7, an upper flange 8, and a lower flange 9. The upper flange 8 and the lower flange 9 are fixed to the top and bottom of the high-pressure chamber body 7, and form a culture chamber inside the high-pressure chamber body 7. A test tube rack is provided inside the culture chamber, and culture test tubes 17 are provided on the test tube rack.

[0022] In a specific embodiment, bolt holes are provided on the upper flange 8 and the lower flange 9 and they are fixed to the upper and lower parts of the high-pressure chamber body 7 by screws, forming a culture chamber between the high-pressure chamber body 7. The culture test tube 17 is inserted into the culture chamber for culture. The culture test tube 17 can be made of polytetrafluoroethylene or glass depending on the actual situation.

[0023] Furthermore, both the upper flange 8 and the lower flange 9 are provided with multiple connection ports 13, which are connected to the culture chamber. The upper flange 8 is provided with a sensor mounting groove 18, which is a blind groove extending along the axial direction of the upper flange.

[0024] In a specific embodiment, the connection port 13 is used for multiple components such as the pressure sensor 10, the constant pressure pump 2, and the drain valve 12, and the sensor mounting slot 18 is used to connect the temperature sensor 11.

[0025] Furthermore, it also includes a pressurization system, which includes a constant pressure pump 2. One end of the constant pressure pump 2 is connected to an explosion-proof valve, and the explosion-proof valve is connected to the connection port 13 on the lower flange 9 to the interior of the culture chamber.

[0026] In a specific implementation, the constant pressure pump 2 pressurizes the inside of the culture chamber, and the pressure is automatically controlled by the PLC to maintain the working pressure of 0-60MPa; the connection port 13 of the upper flange 8 and the constant pressure pump 2 are opened, the pressure value is set, and when water flows out in streams from the outlet of the vent valve 19, the vent valve 19 is closed to apply pressure to the culture chamber, and the pressurization automatically stops after the set pressure is reached.

[0027] Furthermore, it also includes a monitoring system, which includes a temperature sensor 11 and a pressure sensor 10. The temperature sensor 11 is disposed inside the sensor mounting groove 18 of the upper flange, and the pressure sensor 10 is disposed inside the culture chamber through the connection port 13 of the upper flange 8. The temperature sensor 11 and the pressure sensor 10 are connected to a digital display time control instrument. A drain valve 12 is connected to one connection port 13 of the lower flange 9, and a vent valve 19 is connected to one connection port 13 of the upper flange 8.

[0028] In a specific embodiment, temperature sensor 11 is used to monitor the temperature inside the culture chamber, pressure sensor 10 is used to monitor the pressure inside the culture chamber, and the temperature sensor 11 and pressure sensor 10 display the results through a digital display time control instrument. Drain valve 12 and vent valve 19 are used to discharge liquid and gas, respectively.

[0029] Furthermore, it also includes a temperature control system, which includes a water jacket 6, which is disposed on the outside of the culture chamber and is connected to a constant temperature water bath system 16 to form a circulation loop.

[0030] In a specific implementation, the water jacket 6 is used to insulate the culture chamber. The water jacket 6 is made of 316 stainless steel. After the constant temperature water bath system 16 is turned on, liquid enters the water jacket 6 and flows back to the constant temperature water bath system 16 from the outlet of the water jacket 6, forming a circulation. The water jacket 6 keeps the temperature of the culture chamber between -20℃ and 90℃. The water jacket can improve the temperature stability of the culture chamber.

[0031] Furthermore, the water jacket 6 is a spiral circulation structure that surrounds the outside of the culture chamber, and the outermost part of the water jacket 6 is covered with a heat insulation cover.

[0032] In a specific implementation, the water jacket 6 is a spiral structure wound around the outermost part of the culture chamber, and the outermost part of the water jacket 6 is covered with an insulation layer to further improve the insulation effect.

[0033] Furthermore, it also includes a sampling device, which is equipped with a fixing clamp 14 and a lead screw 15. The fixing clamp 14 is controlled by a pneumatic clamping knob to fix the culture tube 17. The lead screw 15 is driven by the control panel to push the piston inside the test tube upward and open the culture tube 17.

[0034] In a specific embodiment, the fixing clamp 14 is located above the sampling device, and the lead screw 15 is located below the fixing clamp 14. The sampling device drives the fixing clamp 14 to fix the culture tube 17 first, and then the lead screw 15 pushes the piston inside the tube to move, opening the culture tube 17.

[0035] Furthermore, a swing-type visualized high-pressure microbial batch culture method includes the following steps: S1. Turn on the constant temperature water bath system 16, set the experimental temperature, and control the temperature of the culture chamber. S2. Sterilize the culture tube 17 used in the experiment, fill the microbial sample in the anaerobic glove box, and put the culture tube 17 onto the test tube rack; S3. Open the upper flange 8 and install the test tube rack and the cushioning cotton for buffering (the cushioning cotton is used to prevent the culture test tube 17 from breaking due to impact), and then re-fix the upper flange 8; S4. Connect the constant pressure pump 2, drain valve 12, pressure sensor 10, temperature sensor 11 and explosion-proof valve, and seal the culture chamber; S5. Open the vent valve 19 of the upper flange 8, set the pressure value and run the constant pressure pump 2. When water flows out in streams at the outlet of the vent valve 19, close the vent valve 19 and apply a constant pressure to the culture chamber to the set pressure. S6. Perform microbial culture and thoroughly mix the sample solution using a shaking device; S7. After the culture experiment is completed, set the constant pressure pump 2 to 0.1MPa and start it. Open the drain valve 12 and the vent valve, and then open the upper flange 8 to take out the test tube rack. S8. Insert the culture tube 17 into the sampling device and clamp it, and push the pistons on both sides of the culture tube 17 to move and pour out the sample.

[0036] Of course, the present invention may have many other embodiments. Based on this embodiment, other embodiments obtained by those skilled in the art without any creative effort are all within the scope of protection of the present invention.

Claims

1. A swing-type visual high-pressure microbial batch culture device, characterized in that, This includes a culture system and a swing system, wherein: The culture system includes a culture chamber with visual observation windows on both sides. The swaying system includes a geared motor, a main support frame, and a digital display time control instrument. The main support frame is used to fix the culture chamber, and the geared motor drives the main support frame to rotate, causing the culture chamber to sway at a uniform speed.

2. The swing-type visual high-pressure microbial batch culture device according to claim 1, characterized in that, The culture chamber includes a high-pressure chamber body, an upper flange, and a lower flange. The upper flange and the lower flange are fixed to the top and bottom of the high-pressure chamber body, respectively, and form a culture cavity inside the high-pressure chamber body. A test tube rack is provided inside the culture cavity, and culture test tubes are placed on the test tube rack.

3. The swing-type visual high-pressure microbial batch culture device according to claim 2, characterized in that, Both the upper flange and the lower flange are provided with multiple connection ports, which are connected to the culture chamber. The upper flange is provided with a sensor mounting slot, which is a blind slot extending along the axial direction of the upper flange.

4. The swing-type visual high-pressure microbial batch culture device according to claim 3, characterized in that, It also includes a pressurization system, which includes a constant pressure pump. One end of the constant pressure pump is connected to an explosion-proof valve, and the explosion-proof valve is connected to the connection port on the lower flange to the inside of the culture chamber.

5. The swing-type visual high-pressure microbial batch culture device according to claim 3, characterized in that, It also includes a monitoring system, which includes a temperature sensor and a pressure sensor. The temperature sensor is installed in the sensor mounting slot of the upper flange, and the pressure sensor is installed inside the culture chamber through the connection port of the upper flange. The temperature sensor and the pressure sensor are connected to a digital display time control instrument. A drain valve is connected to one connection port of the lower flange, and a vent valve is connected to one connection port of the upper flange.

6. The swing-type visual high-pressure microbial batch culture device according to claim 1, characterized in that, It also includes a temperature control system, which includes a water jacket located on the outside of the culture chamber. The water jacket is connected to a constant temperature water bath system and forms a circulation loop.

7. The swing-type visual high-pressure microbial batch culture device according to claim 6, characterized in that, The water jacket is a spiral circulation structure that surrounds the outside of the culture chamber, and the outermost part of the water jacket is covered with a heat insulation cover.

8. The swing-type visual high-pressure microbial batch culture device according to claim 2, characterized in that, It also includes a sampling device, which is equipped with a fixing clamp and a screw. The fixing clamp is controlled by a pneumatic clamping knob to fix the culture tube, and the screw pushes the pistons on both sides of the test tube upward under the drive of the control panel, thereby opening the test tube.

9. A swing-type visual high-pressure microbial batch culture method, characterized in that, The device, applied to the swing-type visual high-pressure microbial batch culture apparatus as described in any one of claims 1-8, includes the following steps: S1. Turn on the water bath system, set the experimental temperature and start the circulation to maintain a constant temperature in the culture chamber; S2. Sterilize the culture tubes used in the experiment, fill the microbial samples in an anaerobic glove box, and install the culture tubes on the test tube rack; S3. Open the upper flange and install the test tube rack and cushioning cotton for buffering, then re-secure the upper flange; S4. Connect the constant pressure pump, drain valve, pressure sensor, temperature sensor and explosion-proof valve, and seal the culture chamber; S5. Open the vent valve on the upper flange, set the pressure value and run the constant pressure pump. When water flows out in streams at the outlet of the vent valve, close the vent valve and apply a constant pressure to the culture chamber to the set pressure. S6. Perform microbial culture and thoroughly mix the sample solution using a shaking device; S7. After the culture experiment is completed, set the constant pressure pump to 0.1MPa and start it. Open the drain valve and the vent valve, and then open the upper flange to take out the test tube rack. S8. Place the culture tube into the sampling device and clamp it, then push the pistons on both sides of the culture tube to move and pour out the sample.

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