Small low-temperature plasma microorganism mutation breeding device

Abstract

The invention provides a small low-temperature plasma microorganism mutation breeding device which comprises a shell, a mutagenesis cavity is formed in the shell, a sample carrying disc is rotationally arranged in the mutagenesis cavity and driven to rotate through a servo motor arranged in the mutagenesis cavity, a centrifugal tube circular clamping support is further arranged below the sample carrying disc, and the centrifugal tube circular clamping support is arranged in the shell. A centrifugal tube circular clamping support is arranged in the mutagenesis cavity and used for clamping a centrifugal tube, an ultrasonic washing tray is arranged below the centrifugal tube circular clamping support, a composite discharge electrode is arranged above the sample carrying tray in the mutagenesis cavity, and an ultraviolet lamp tube is further arranged in the mutagenesis cavity. According to the method, the plasma with higher intensity and higher density can be generated at normal temperature and normal pressure, the assistance of conditions such as a vacuum environment and rare gas is not needed, continuous mutagenesis can be carried out on a large scale, and the mutagenesis success rate is higher; harmful gas generated in an experiment can be filtered and degraded through the arranged composite filtering module, and then the harmful gas is discharged after reaching a concentration range acceptable by a human body, so that the probability of bacterial contamination is reduced.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial breeding devices, and in particular relates to a small low-temperature plasma microbial mutation breeding device. Background Art

[0002] In the modern bio-industry, high-quality microorganisms occupy a core position. High-quality microorganisms have broad application prospects in various fields. They can not only solve many practical problems, but also provide important technical support for achieving sustainable development. How to use efficient biological mutagenesis technology to achieve the optimization and iteration of microorganisms has always been a difficult problem. Conventional biological mutagenesis methods have various problems, such as high randomness and low efficiency of mutagenesis, environmental pollution of chemical mutagenesis reagents and health problems for users, physical mutagenesis requires expensive equipment and is easy to produce tolerance, and some genetic engineering biotechnology and other mutagenesis methods have complex processes, high technical barriers, and difficult operations. They are even restricted by ethics and regulations, and may cause ecological and safety problems. Therefore, conventional mutagenesis methods can no longer adapt to the development of modern industrial microorganisms. There is an urgent need for a new mutagenesis breeding method with high mutagenesis efficiency, low technical barriers, green environmental protection, and no need for expensive equipment.

[0003] Plasma is the fourth state of matter, consisting of a large number of free electrons, ions and neutral particles. It forms plasma by applying high energy to the gas to ionize gas atoms or molecules. Plasma contains a variety of high-energy particles, such as free radicals, electrons, ions and ultraviolet photons. These particles are highly active and can interact with biological molecules to trigger a series of chemical and physical reactions. When high-energy particles in plasma come into contact with microbial cells, they damage the DNA in the cells. After the DNA is damaged, the cells will start their own repair mechanism. However, the repair process is not always accurate and may introduce errors, leading to gene mutations. These mutations may change the physiological characteristics of microorganisms and produce strains with excellent characteristics. After plasma treatment, the microbial population will produce a large number of gene mutations. After screening and cultivation, high-quality strains can be obtained. Plasma mutagenesis breeding technology overcomes many shortcomings of traditional methods and has the advantages of high efficiency, safety, non-contact, non-genetic modification, wide applicability, low-temperature operation, automation and diversity. It is a new microbial breeding technology with great potential.

[0004] At present, the application of plasma technology in biological breeding is still rare. The reasons are as follows: plasma is difficult to form at room temperature and pressure, and special conditions such as high pressure and high temperature are required; biological experiments are particular, and mutagenesis experiments in the external environment will easily cause the samples to be contaminated with bacteria, resulting in experimental failure; only a few experimental samples can be added each time for mutagenesis, and too many operations of putting in and taking out each time the mutagenesis is completed are extremely easy to be infected by external impurities, resulting in overall experimental failure; the current plasma is generated under air conditions, and due to the presence of high-energy electrons, it collides with O2, N2 and other substances in the air to form ozone, nitrogen oxides, etc. Chemicals, superoxides and hydroxides, etc. First of all, these substances will irritate the human respiratory system, causing symptoms such as coughing, sore throat, shortness of breath and decreased lung function, and will irritate the skin and eyes, which is extremely harmful to the user. Secondly, high concentrations of harmful gases will destroy the experimental environment and even kill the induced cells. The plasma mutagenesis devices currently on the market generally have the need for heat dissipation and a rare gas environment, which means that the equipment also needs external water cooling, air cooling, gas cylinders, air intake devices and other peripherals, which makes the whole set of equipment large in size and difficult to use.

[0005] Based on this, a small-scale low-temperature plasma microbial mutagenesis breeding device was proposed. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide a small low-temperature plasma microbial mutation breeding device in view of the deficiencies of the above-mentioned prior art to solve the problems raised in the above-mentioned background technology.

[0007] In order to solve the above technical problems, the technical solution adopted by the present invention is: a small low-temperature plasma microbial mutation breeding device, comprising a shell, wherein a mutagenesis cavity is arranged in the shell;

[0008] A sample carrier plate is rotatably arranged in the mutagenesis chamber, and the sample carrier plate is driven to rotate by a servo motor arranged in the mutagenesis chamber. A circular centrifuge tube holder is also arranged below the sample carrier plate, and the circular centrifuge tube holder is used to clamp the centrifuge tube. An ultrasonic washing tray is arranged below the circular centrifuge tube holder, and an ultrasonic elution vibrator is arranged on the bottom surface of the ultrasonic washing tray.

[0009] A composite discharge electrode is arranged above the sample carrier plate in the mutagenesis chamber, and an ultraviolet lamp is also arranged in the mutagenesis chamber, and the disinfection operation is completed by the ultraviolet lamp.

[0010] As a further explanation of the present invention, a vent is provided below the mutagenesis chamber, a filter element is installed in the middle layer of the vent, a ducted vortex fan is connected above the mutagenesis chamber, the air inlet of the ducted vortex fan is connected to the mutagenesis chamber, the air outlet of the ducted vortex fan is connected to the exhaust port, and a composite filter module is provided in the exhaust port.

[0011] As a further illustration of the present invention, the composite discharge electrode is a columnar structure with a square protrusion at the tip, and the composite discharge electrode is connected to a high-voltage module, and the high-voltage module is used to provide a high-voltage point to generate plasma.

[0012] As a further explanation of the present invention, the sample carrier plate is a circular plate with a diameter of 100 mm, on which ten cones are evenly distributed. There is a groove with a size of 10 mm in the middle of the cone, which can be inserted with a circular stainless steel sheet with a diameter of 10 mm during the experiment. The circular stainless steel sheet can be loaded with the bacterial solution that needs to be induced to mutate.

[0013] As a further illustration of the present invention, PBS buffer for eluting bacteria is added to the centrifuge tube, and after the loading plate buckle on the loading plate round table is triggered and released, the circular stainless steel sheet falls into the centrifuge tube. After the centrifuge tube is manually closed, the circular clip of the centrifuge tube is pressed down into the ultrasonic washing tray to elute the induced bacteria from the circular stainless steel sheet.

[0014] As a further illustration of the present invention, the servo motor drives the sample carrier plate to rotate during the experiment. After completing the mutagenesis of the bacterial solution in one stainless steel sheet, the servo motor will rotate to the next sample to continue the mutagenesis, so that the plasma generated by the composite discharge electrode can evenly treat each circular stainless steel sheet.

[0015] As a further illustration of the present invention, a microprocessor, a position sensor, a temperature sensor, a humidity sensor and a harmful gas concentration sensor are also arranged in the mutagenesis chamber;

[0016] Among them, the microprocessor is respectively connected with the position sensor, temperature sensor, humidity sensor and harmful gas concentration sensor by signal, the position sensor is connected with the servo motor, the temperature sensor, humidity sensor and harmful gas concentration sensor are all fixed in the mutagenesis chamber, the temperature sensor and humidity sensor are used to detect the temperature and humidity environment in the mutagenesis chamber, and the gas concentration sensor detects the harmful gas in the mutagenesis chamber in real time.

[0017] As a further explanation of the present invention, a perspective window, a display screen 2, operation buttons and a switch key are also provided on the outside of the shell, and an incandescent lamp tube for lighting is also provided inside the shell.

[0018] Compared with the prior art, the present invention has the following advantages:

[0019] 1. The present invention can generate plasma with high intensity and high density at normal temperature and pressure, without the assistance of vacuum environment and rare gas conditions, and can carry out continuous mutagenesis in large quantities with high mutagenesis success rate. The harmful gases generated in the experiment can be filtered and degraded through the composite filtration module and discharged after reaching the concentration range acceptable to the human body, thereby reducing the probability of bacterial infection.

[0020] 2. The device of the present invention is small in size and has low power, especially the core part of the device, which is a high-voltage power supply for generating plasma, and has a power of no more than 50 watts. There is no need to add bulky air-cooling, water-cooling or other heat dissipation devices. The device is simple and easy to operate. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is a schematic diagram of the overall structure of the present invention;

[0022] Figure 2 It is a schematic diagram of the internal structure of the present invention;

[0023] Figure 3 It is a schematic diagram of the installation of the duct type vortex fan of the present invention;

[0024] Figure 4 It is a schematic diagram of the installation of the filter element of the present invention;

[0025] Figure 5 It is a schematic diagram of the installation of the composite filtration module of the present invention;

[0026] Figure 6 It is a schematic diagram of the structure of the object carrier disk truncated table of the present invention.

[0027] Description of reference numerals:

[0028] 1-perspective window; 2-display screen; 3-operation button; 4-power button; 5-housing; 6-ultraviolet lamp tube; 7-incandescent lamp tube; 8-compound discharge electrode; 9-sample carrier plate; 10-circular holder for centrifuge tube; 11-centrifuge tube; 12-ultrasonic elution plate; 13-ultrasonic elution vibrator; 14-high-voltage module; 15-servo motor; 16-gas pipeline; 17-filter element; 18-compound filter module; 19-carrier plate round table; 20-carrier plate load plate buckle; 21-ducted vortex blower. DETAILED DESCRIPTION

[0029] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] like Figure 1-6 As shown, the present invention provides a technical solution: a small low-temperature plasma microbial mutation breeding device, comprising a shell 5, wherein a mutagenesis cavity is arranged in the shell 5;

[0031] A sample carrier plate 9 is rotatably arranged in the mutagenesis chamber, and the sample carrier plate 9 is driven to rotate by a servo motor 15 arranged in the mutagenesis chamber. A circular centrifuge tube holder 10 is also arranged below the sample carrier plate 9, and the circular centrifuge tube holder 10 is used to clamp a centrifuge tube 11. An ultrasonic washing tray 12 is arranged below the circular centrifuge tube holder 10, and an ultrasonic elution vibrator 13 is arranged on the bottom surface of the ultrasonic washing tray 12.

[0032] The sample carrier plate 9 is a disc with a diameter of 100 mm, on which ten cones are evenly distributed. There is a groove with a size of 10 mm in the middle of the cone, into which a circular stainless steel sheet with a diameter of 10 mm can be inserted during the experiment, and the bacterial solution that needs to be induced can be loaded on the circular stainless steel sheet.

[0033] PBS buffer for eluting bacteria is added to the centrifuge tube 11. After the carrier plate loading sheet buckle 20 on the carrier plate truncated table 19 is triggered and released, the circular stainless steel sheet falls into the centrifuge tube 11. After the centrifuge tube 11 is manually closed, the circular centrifuge tube holder 10 is pressed down into the ultrasonic washing tray 12 to elute the induced bacteria from the circular stainless steel sheet.

[0034] The servo motor 15 drives the sample carrier plate 9 to rotate during the experiment. After completing the mutagenesis of the bacterial solution in one stainless steel sheet, the servo motor 15 will rotate to the next sample to continue the mutagenesis, so that the plasma generated by the composite discharge electrode 8 can evenly process each circular stainless steel sheet.

[0035] A composite discharge electrode 8 is arranged above the sample carrier plate 9 in the mutagenesis chamber. The composite discharge electrode 8 is a columnar structure with a square protrusion at the tip. The composite discharge electrode 8 is connected to a high-voltage module 14, and the high-voltage module 14 is used to provide a high-voltage point to generate plasma.

[0036] An ultraviolet lamp 6 is also provided in the mutagenesis chamber, and the disinfection operation is completed by the ultraviolet lamp 6.

[0037] A vent is provided below the mutagenesis chamber, a filter element 17 is installed in the middle layer of the vent, a duct-type vortex fan 21 is connected above the mutagenesis chamber, an air inlet of the duct-type vortex fan 21 is connected to the mutagenesis chamber, an air outlet of the duct-type vortex fan 21 is connected to an exhaust port, and a composite filter module 18 is provided in the exhaust port.

[0038] The mutagenesis chamber is also provided with a microprocessor, a position sensor, a temperature sensor, a humidity sensor and a harmful gas concentration sensor;

[0039] Among them, the microprocessor is respectively connected with the position sensor, temperature sensor, humidity sensor and harmful gas concentration sensor by signal, the position sensor is connected with the servo motor, the temperature sensor, humidity sensor and harmful gas concentration sensor are all fixed in the mutagenesis chamber, the temperature sensor and humidity sensor are used to detect the temperature and humidity environment in the mutagenesis chamber, and the gas concentration sensor detects the harmful gas in the mutagenesis chamber in real time.

[0040] The outer side of the housing 5 is also provided with a perspective window 1, a display screen 2, an operation button 3 and a switch key 4, and the inside of the housing 5 is also provided with an incandescent lamp tube 7 for lighting.

[0041] In summary, when in use, the device is started, the display screen 2 lights up, and the ultraviolet lamp 6 in the mutagenesis chamber is started to disinfect the mutagenesis chamber by ultraviolet irradiation for 30 minutes to ensure a sterile environment in the chamber. After the disinfection is completed, the ultraviolet lamp 6 will automatically turn off.

[0042] Then use a pipette to evenly distribute 10 microliters or 20 microliters of the prepared microbial bacterial solution on the stainless steel load sheet. The surface of the stainless steel load sheet has an arc, and the surface tension will prevent the loaded bacterial solution from leaking out. After the bacterial solution of the stainless steel load sheet is dried or nearly dried, the stainless steel load sheet is placed in the depression of the loading plate truncated table 19 and stuck to ensure that the sample is firmly fixed and will not move or tilt during the experiment. Prepare ten sterilized centrifuge tubes 11 with a capacity of 5 ml, open the centrifuge tube cover, add an appropriate amount of PBS buffer for subsequent elution of the sample bacterial solution, and then manually insert it into the centrifuge tube circular card holder 10 below the sample loading plate 9, adjust the position, and make it close to the bottom surface of the sample loading plate 9, so that the loading plate loading plate buckle 20 can collect the sample of the stainless steel load sheet into the centrifuge tube 11 after the experiment is completed, and then add sterile water to the ultrasonic elution plate 12 below for ultrasonic elution, which is used for subsequent ultrasonic elution of the bacterial solution.

[0043] Next, the mutagenesis mode required for the experiment is set by operating the button 3 in conjunction with the microprocessor, such as continuous mutagenesis for 5 seconds, 10 seconds, or mutagenesis for 5 seconds followed by a 5-second interval and then mutagenesis for 10 seconds, such as a power value of 10 watts or 12 watts, etc., to ensure that the mode is appropriate. Different microorganisms have different requirements for the required plasma mutagenesis, and appropriate parameters are set according to specific experimental requirements. For example, for some bacteria, 10 watts of mutagenesis for 30 seconds will achieve a lethality of 90%, which is the best mutagenesis time, while for some molds, it may be necessary to control it at about 50 seconds and 12 watts to achieve the best mutagenesis effect. After the mutagenesis mode and parameters are set, the high-voltage module 14 is started, and the composite discharge electrode 8 begins to generate glow discharge plasma in the mutagenesis chamber, and the plasma contacts the bacterial solution on the stainless steel load sheet to start the mutagenesis process. Low-temperature glow plasma has high-energy electrons and active particles, which can interact with the DNA of microorganisms to cause gene mutations.

[0044] 1-perspective window; 2-display screen; 3-operation button; 4-power button; 5-housing; 6-ultraviolet lamp tube; 7-incandescent lamp tube; 8-compound discharge electrode; 9-sample carrier plate; 10-circular holder for centrifuge tube; 11-centrifuge tube; 12-ultrasonic elution plate; 13-ultrasonic elution vibrator; 14-high-voltage module; 15-servo motor; 16-gas pipeline; 17-filter element; 18-compound filter module; 19-carrier plate round table; 20-carrier plate load plate buckle; 21-ducted vortex blower.

[0045] During the mutagenesis process, the servo motor 15 controls the rotation of the sample carrier plate 9 so that the bacterial solution on each stainless steel carrier plate is uniformly exposed to the plasma. By adjusting the distance between the sample carrier plate and the composite discharge electrode 8, the discharge state can be optimized, thereby improving the mutagenesis efficiency.

[0046] For example, when performing bacterial mutagenesis, the sample tray 9 can be adjusted to a position closer to the composite discharge electrode 8 to enhance the effect of plasma. For some microorganisms that are more sensitive to environmental changes, the distance between the sample tray 9 and the composite discharge electrode 8 can be appropriately adjusted to reduce damage to the microorganisms. After each stainless steel load sheet is processed, the position sensor detects the position of the sample tray 9 and sends a signal to the microprocessor to control the servo motor 15 to rotate to the next sample.

[0047] After the mutagenesis is completed, the system will detect the concentration of harmful gases in the mutagenesis chamber. When the concentration exceeds the threshold that the human body can tolerate, the ducted vortex fan 21 will be started to extract the gas in the mutagenesis chamber. After being processed by the composite filtration module, it reaches the cleanliness standard and is discharged into the air. The concentration of harmful gases in the chamber is within the threshold that the human body can tolerate. At this time, the device door can be opened freely, and the loading plate load sheet buckle is manually triggered to make the mutated stainless steel loading plate fall into the centrifuge tube 11 below which is pre-filled with PBS buffer. After covering the centrifuge tube 11, the circular holder 10 of the centrifuge tube is pressed down to the ultrasonic elution plate 12 filled with sterile water, the chamber door of the mutagenesis chamber is closed, and the ultrasonic elution mode is set. The duration can be selected, such as 30 seconds, 60 seconds, etc., and the ultrasonic elution vibrator 13 is used to control the ultrasonic elution plate 12 to perform the elution operation.

[0048] After elution, the sample is diluted to an appropriate concentration, and a culture medium suitable for the microorganism is prepared. It is evenly spread on the culture medium with a spreading stick, and its growth, morphological changes and mutation rate are recorded to complete the processing.

[0049] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0050] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A small low-temperature plasma microbial mutation breeding device, characterized in that: It comprises a housing (5), wherein a mutagenesis cavity is arranged in the housing (5); A sample carrier plate (9) is rotatably arranged in the mutagenesis chamber, and the sample carrier plate (9) is driven to rotate by a servo motor (15) arranged in the mutagenesis chamber. A circular centrifuge tube holder (10) is also arranged below the sample carrier plate (9), and the circular centrifuge tube holder (10) is used to clamp a centrifuge tube (11). An ultrasonic washing tray (12) is arranged below the circular centrifuge tube holder (10), and an ultrasonic elution vibrator (13) is arranged on the bottom surface of the ultrasonic washing tray (12); A composite discharge electrode (8) is arranged above the sample carrier plate (9) in the mutagenesis chamber, and an ultraviolet lamp tube (6) is also arranged in the mutagenesis chamber, and disinfection is performed by the ultraviolet lamp tube (6).

2. A small low-temperature plasma microbial mutation breeding device according to claim 1, characterized in that: A vent is provided below the mutagenesis chamber, a filter element (17) is installed in the middle layer of the vent, a duct-type vortex fan (21) is connected to the top of the mutagenesis chamber, an air inlet of the duct-type vortex fan (21) is connected to the mutagenesis chamber, an air outlet of the duct-type vortex fan (21) is connected to an exhaust port, and a composite filter module (18) is provided in the exhaust port.

3. A small low-temperature plasma microbial mutation breeding device according to claim 1, characterized in that: The composite discharge electrode (8) is a columnar structure with a square protrusion at the tip. The composite discharge electrode (8) is connected to a high-voltage module (14). The high-voltage module (14) is used to provide a high-voltage point to generate plasma.

4. A small low-temperature plasma microbial mutation breeding device according to claim 1, characterized in that: The sample carrier plate (9) is a circular plate with a diameter of 100 mm. Ten truncated cones are evenly distributed on the circular plate. A groove with a size of 10 mm is provided in the middle of the truncated cone. During the experiment, a circular stainless steel sheet with a diameter of 10 mm can be inserted. The bacterial liquid that needs to be induced can be loaded on the circular stainless steel sheet.

5. A small low-temperature plasma microbial mutation breeding device according to claim 4, characterized in that: PBS buffer for eluting bacteria is added to the centrifuge tube (11), and after the carrier plate loading sheet buckle (20) on the carrier plate truncated table (19) is triggered and released, the circular stainless steel sheet falls into the centrifuge tube (11), and after the centrifuge tube (11) is manually closed, the circular centrifuge tube holder (10) is pressed down into the ultrasonic washing tray (12) to elute the induced bacteria from the circular stainless steel sheet.

6. A small low-temperature plasma microbial mutation breeding device according to claim 5, characterized in that: The servo motor (15) drives the sample carrier plate (9) to rotate during the experiment. After completing the mutagenesis of the bacterial solution in one stainless steel sheet, the servo motor (15) will rotate to the next sample to continue the mutagenesis, so that the plasma generated by the composite discharge electrode (8) can be evenly processed to each circular stainless steel sheet.

7. A small low-temperature plasma microbial mutation breeding device according to claim 1, characterized in that: The mutagenesis chamber is also provided with a microprocessor, a position sensor, a temperature sensor, a humidity sensor and a harmful gas concentration sensor; Among them, the microprocessor is respectively connected with the position sensor, temperature sensor, humidity sensor and harmful gas concentration sensor by signal, the position sensor is connected with the servo motor, the temperature sensor, humidity sensor and harmful gas concentration sensor are all fixed in the mutagenesis chamber, the temperature sensor and humidity sensor are used to detect the temperature and humidity environment in the mutagenesis chamber, and the gas concentration sensor detects the harmful gas in the mutagenesis chamber in real time.

8. The small low-temperature plasma microbial mutation breeding device according to claim 1, characterized in that: The outer side of the housing (5) is also provided with a perspective window (1), a display screen (2), an operation button (3) and a switch key (4), and the inside of the housing (5) is also provided with an incandescent lamp tube (7) for lighting.

Images