Continuous separation device and method for marine microorganisms in high-pressure environment

Abstract

The invention discloses a continuous separation device and method for marine microorganisms in a high-pressure environment, and relates to the technical field of online separation of marine microorganisms. The method comprises the steps that a microorganism enrichment unit is arranged for microorganism enrichment, and a nutrient solution needed by culture is provided through a multi-sequence nutrient solution supply unit; the gas injection unit is used for providing culture gas or inert gas required by culture, so that the pressure in the microorganism enrichment unit is consistent with the pressure value of microorganisms in the ocean in situ, and the effectiveness of enrichment culture is ensured; after the enrichment culture is performed for a period of time, pumping the microbial bacterial liquid into a multi-stage separation unit, and completing multi-stage separation of cells with different sizes in the microbial bacterial liquid, so as to obtain target microbial cells; and finally, transferring the target microbial cells to a monitoring culture unit for culture and fixation, and carrying out cell characteristic detection by utilizing an external multi-omics unit. The device can continuously separate marine microorganisms with different sizes in a marine in-situ high-pressure environment and then carry out characteristic detection, is simple to operate, improves the separation efficiency, is accurate in detection result, and fits the actual environment.

Description

Technical Field

[0001] The present invention relates to the technical field of online separation of marine microorganisms, and more specifically, to a device and method for continuous separation of marine microorganisms under a high pressure environment. Background Art

[0002] Marine microorganisms are simple organisms that grow and reproduce in the marine environment, are tiny in size, single-celled or multi-celled with relatively simple individual structures, or even have no cell structure. The ocean where marine microorganisms live is the largest and oldest habitat on Earth, occupying 71% of the Earth's area and with an average depth of 3.8 km. Due to the uneven distribution of land and sea on Earth, the marine habitat is actually composed of many different macrohabitats, and different habitat conditions breed different microbial populations.

[0003] At present, microbial enrichment and separation experiments have been carried out under high-pressure and normal-pressure environments. Marine microorganisms are regulated by pressure factors. Enrichment and separation under normal-pressure environments can only obtain some pressure-resistant microorganisms, and cannot obtain the original unique indigenous microorganisms of the deep sea. However, after enriching and culturing microorganisms under high-pressure environments, they are generally separated by streaking, limiting dilution, etc. In addition, if the characteristics of microorganisms are to be detected, such as gene transcription of microorganisms, the environmental pressure needs to be changed, and the results obtained are greatly deviated from the actual environment. Summary of the invention

[0004] In order to overcome the defects of the above-mentioned existing high-pressure enrichment and cultivation process of marine microorganisms, which requires pressure release to separate and detect the characteristics of microorganisms of different sizes, and the inaccurate detection results, the present invention provides a continuous separation device and method for marine microorganisms under a high-pressure environment. The device and method can continuously separate marine microorganisms of different sizes under a high-pressure environment and then perform characteristic detection. The device and method have simple operation, improved separation efficiency, accurate detection results, and are in line with the actual environment.

[0005] In order to solve the above technical problems, the technical solution of the present invention is as follows:

[0006] The present invention discloses a continuous separation device for marine microorganisms under high pressure environment, comprising: a microorganism enrichment unit, a gas injection unit, a multi-sequence nutrient solution supply unit, a multi-stage separation unit, a monitoring and cultivation unit and a central control unit;

[0007] The air inlet end of the microorganism enrichment unit is connected to the air outlet end of the gas injection unit, and the liquid inlet end of the microorganism enrichment unit is connected to the liquid outlet end of the multi-sequence nutrient solution supply unit;

[0008] The input end of the multi-stage separation unit is connected to the liquid outlet end of the microorganism enrichment unit, and the output end of the multi-stage separation unit is connected to the input end of the monitoring and cultivation unit;

[0009] The data output ends of the microorganism enrichment unit and the monitoring and cultivation unit are connected to the data input end of the central control unit, and the control ends of the microorganism enrichment unit, the gas injection unit, the multi-sequence nutrient solution supply unit, the multi-stage separation unit, and the monitoring and cultivation unit are connected to the control end of the central control unit.

[0010] The present invention sets up a microorganism enrichment unit to enrich microorganisms, uses a multi-sequence nutrient solution supply unit to provide the microorganism enrichment unit with nutrient solution required for microorganism culture, uses a gas injection unit to provide the microorganism enrichment unit with culture gas or inert gas required for microorganism culture, and makes the pressure in the microorganism enrichment unit consistent with the pressure value of the microorganism in situ in the ocean, thereby ensuring the effectiveness of enrichment culture; after a period of enrichment culture, the microorganism liquid is pumped into a multi-stage separation unit to complete the multi-stage separation of cells of different sizes of the microorganism liquid to obtain target microorganism cells; finally, the target microorganism cells are transferred to a monitoring culture unit to culture and fix the target microorganism, and the cell characteristics are detected using an external multi-omics unit.

[0011] Preferably, the multi-stage separation unit comprises a first filter, a microorganism separator, a second filter, a first separation filter, a second separation filter and a multi-channel single cell sorter;

[0012] The first filter, the microorganism separator, the second filter, and the first separation filter are connected in sequence; the first output end of the first separation filter is connected to the first input end of the multi-channel single cell sorter, the second output end of the first separation filter is connected to the input end of the second separation filter, the first output end of the second separation filter is connected to the second input end of the multi-channel single cell sorter, and the second output end of the second separation filter is connected to the third input end of the multi-channel single cell sorter;

[0013] The input end of the first filter serves as the input end of the multi-stage separation unit and is connected to the liquid outlet end of the microorganism enrichment unit. The first output end, the second output end, and the third output end of the multi-channel single cell sorter all serve as the output ends of the multi-stage separation unit and are connected to the input end of the monitoring and culture unit.

[0014] The microbial solution is pumped into the first filter to filter out large particles of impurities to obtain a first-stage filtered microbial solution, which enters the microbial separator for dilution and separation to obtain a separated diluted microbial solution; the separated diluted microbial solution enters the second filter to filter out sediment to obtain a second-stage filtered microbial solution; the filter diameter of the first separation filter is larger than the filter diameter of the second separation filter, the second-stage filtered microbial solution first enters the first separation filter for first-stage separation, the microbial solution that does not pass through enters the first input end of the multi-channel single cell sorter through the first output end of the first separation filter; the microbial solution that passes through enters the second filter through the second output end of the first separation filter. The filter performs secondary separation, the microbial solution that does not pass through enters the second input end of the multi-channel single cell sorter through the first output end of the second separation filter, and the microbial solution that passes through enters the third input end of the multi-channel single cell sorter through the second output end of the second separation filter, completing the multi-stage separation of cells of different sizes of the microbial solution; after entering the multi-channel single cell sorter, the state and type of the single cells entering from each input end are observed through the window on the multi-channel single cell sorter to determine whether the target microbial cells are obtained. If the target microbial cells are not obtained, the microbial solution of the first-stage filtration in the microbial separator is updated, and the above steps are repeated until the target microbial cells are obtained.

[0015] Preferably, the first filter comprises a first filter frame and a first filter membrane, and the surface of the first filter frame is covered with the first filter membrane;

[0016] The second filter comprises a second filter frame and a second filter membrane, and the surface of the second filter frame is covered with the second filter membrane;

[0017] The filtration diameter of the second filtration membrane is smaller than the filtration diameter of the first filtration membrane.

[0018] The first filter is used to filter out large particle impurities, such as humus, in the process of enrichment and cultivation of microbial liquid, and the second filter is used to filter out sediments larger than the diameter of the microorganisms in the separated diluted microbial liquid.

[0019] Preferably, the first separation filter comprises a first protective shell and a first filter screen, and the first filter screen is arranged inside the first protective shell;

[0020] The second separation filter comprises a second protective shell and a second filter screen, and the second filter screen is arranged inside the second protective shell;

[0021] The filtering diameter of the second filter net is smaller than the filtering diameter of the first filter net.

[0022] The first separation filter and the second separation filter are used to separate cells of different diameters. Since the filtration diameter of the second filter is smaller than the filtration diameter of the first filter, the microbial solution of the secondary filtration first enters the first separation filter for primary separation. The cell size of the microbial solution that has not passed through the first filter is the largest and enters the multi-channel single cell sorter through the first output end of the first separation filter; the microbial solution that has passed through the first filter enters the second separation filter through the second output end of the first separation filter for secondary separation. The cell size of the microbial solution that has not passed through the second filter is the second largest and enters the multi-channel single cell sorter through the first output end of the second separation filter; the cell size of the microbial solution that has passed through the second filter is the smallest and enters the multi-channel single cell sorter through the second output end of the second separation filter.

[0023] Preferably, the microorganism separator comprises a separation chamber, a diluent storage chamber, a filtration pump, a magnet and magnetic beads;

[0024] The magnetic beads are arranged inside the separation chamber, and the magnet is arranged on the lower surface of the separation chamber, corresponding to the position of the magnetic beads;

[0025] The water outlet end of the suction filter pump is suspended in the air, the water inlet end of the suction filter pump extends into the interior of the separation bin, and the control end of the suction filter pump is connected to the control end of the central control unit;

[0026] The diluent storage bin is provided with diluent, and the diluent storage bin is connected to the upper surface of the separation bin through a pipeline.

[0027] After the primary filtered microbial solution enters the separation chamber in the microbial separator, the diluent storage chamber is opened, and the diluent enters the separation chamber to dilute the primary filtered microbial solution according to a preset ratio to obtain the diluted microbial solution. The diluent is mainly used to dilute the solution and protect the cells from damage, and can provide certain nutrients; the function of the magnet and magnetic beads is to drive the diluted microbial solution in the separation chamber to rotate, separate the sediment and the cell-bound part, and obtain the separated microbial diluted solution. The function of the suction pump is to use the suction pump to extract and discharge the primary filtered microbial solution in the separation chamber when the target microbial cells are not obtained in the multi-channel single cell sorter, update the primary filtered microbial solution in the separation chamber, and perform the separation process again.

[0028] Preferably, the monitoring culture unit comprises a culture chamber, a support frame, a plurality of culture cups, a handle, an environmental indicator sensor, a first injection pump and a fixing liquid container;

[0029] The environmental indicator sensor, the support frame and the plurality of culture cups are arranged in the culture chamber, and the data output end of the environmental indicator sensor is connected to the data input end of the central control unit;

[0030] The plurality of culture cups are evenly arranged on the upper surface of the support frame along the circumference; the handle is arranged vertically, one end of which is connected to the lower surface of the support frame, and the other end of which extends from the bottom surface of the culture chamber;

[0031] The upper surface of the culture chamber is provided with a first through hole and a second through hole, and the positions of the first through hole and the second through hole correspond to any two culture cups;

[0032] The first output end, the second output end and the third output end of the multi-channel single cell sorter are connected to the first through hole; the output end of the fixed liquid container is connected to one end of the first injection pump, and the other end of the first injection pump is connected to the second through hole.

[0033] Preferably, the environmental indicator sensor includes an environmental pressure sensor, an environmental temperature sensor and a gas concentration detector;

[0034] The culture chamber is cylindrical, the support frame is circular, and several culture cups are evenly arranged on the upper surface of the support frame along the circumference. The handle is vertically connected to the lower surface of the support frame. The upper surface of the culture chamber is provided with a first through hole and a second through hole, which correspond to any two culture cups respectively. When in use, the output end of the multi-channel single-cell sorter corresponding to the target microbial cells is opened, and enters the culture cup through the first through hole for culture; the handle is turned to make the next culture cup correspond to the first through hole; after a period of culture, the handle is turned to make the culture cup containing the cultured target microbial cells correspond to the second through hole, and the first injection pump is controlled to start so that the fixative enters the culture cup to complete the fixation of the cultured target microbial cells and obtain a fixed sample.

[0035] Preferably, the microorganism enrichment unit comprises a microorganism enrichment kettle, a magnetic stirrer, a vent valve, a liquid inlet valve, a pressure sensor, a temperature sensor, a water bath and an air inlet valve;

[0036] The side wall of the microorganism enrichment kettle is provided with a liquid outlet, and the liquid outlet serves as the liquid outlet of the microorganism enrichment unit and is connected to the input end of the first filter;

[0037] The microorganism enrichment kettle is arranged in a water bath, a magnetic stirrer is arranged on the lower surface of the microorganism enrichment kettle, and the control ends of the water bath and the magnetic stirrer are connected to the control end of the central control unit;

[0038] One end of the vent valve is connected to the upper surface of the microorganism enrichment kettle, the other end of the vent valve is suspended in the air, and the control end of the vent valve is connected to the control end of the central control unit;

[0039] One end of the air inlet valve is connected to the upper surface of the microorganism enrichment kettle, the other end of the air inlet valve is connected to the gas outlet end of the gas injection unit as the air inlet end of the microorganism enrichment unit, and the control end of the air inlet valve is connected to the control end of the central control unit;

[0040] One end of the liquid inlet valve is connected to the side wall of the microorganism enrichment kettle, the other end of the liquid inlet valve is connected to the liquid outlet of the multi-sequence nutrient solution supply unit as the liquid inlet end of the microorganism enrichment unit, and the control end of the liquid inlet valve is connected to the control end of the central control unit;

[0041] The temperature sensor and the pressure sensor are both arranged on the upper surface of the microorganism enrichment kettle, and the data output ends of the temperature sensor and the pressure sensor are both connected to the data input end of the central control unit.

[0042] The upper surface of the microbial enrichment kettle is detachable, which is convenient for sterilization and placement of substrates to be cultured; a magnetic stirrer is set on the lower surface to enhance the mass transfer process in the microbial enrichment kettle and increase the energy and nutrient utilization efficiency of the microbial culture. The pressure sensor and temperature sensor are used to monitor the pressure and temperature changes in the microbial enrichment kettle in real time, the vent valve is used to reduce the real-time pressure in the microbial enrichment kettle, and the water bath is used to maintain the constant temperature in the microbial enrichment kettle. The air inlet valve and liquid inlet valve are used to control the entry of culture gas or inert gas and nutrient solution respectively, so that the microbial culture is always in the best growth environment.

[0043] Preferably, the gas injection unit comprises a booster pump, an air compressor, a gas storage tank and a pressure regulating valve connected in sequence;

[0044] The other end of the pressure regulating valve is connected to the other end of the air inlet valve as the outlet end of the gas injection unit; the control end of the booster pump, the air compressor and the pressure regulating valve is connected to the control end of the central control unit;

[0045] The gas storage tank is filled with culture gas or inert gas.

[0046] Preferably, the multi-sequence nutrient solution supply unit comprises a plurality of nutrient bottles, a plurality of liquid outlet valves and a second liquid injection pump;

[0047] The liquid outlet of each nutrient bottle is connected to one end of a liquid outlet valve, and the other end of each liquid outlet valve is connected to one end of a second liquid injection pump. The other end of the second liquid injection pump serves as the liquid outlet end of the multi-sequence nutrient solution supply unit and is connected to the other end of the liquid inlet valve; the control end of each liquid outlet valve and the control end of the second liquid injection pump are connected to the control end of the central control unit;

[0048] A nutrient solution is arranged in each of the nutrient bottles.

[0049] The present invention also provides a method for continuous separation of marine microorganisms under high pressure, which is applied to the above-mentioned continuous separation device, comprising:

[0050] S1: sterilizing the microorganism enrichment unit, the multi-stage separation unit and the monitoring culture unit; adding the substrate to be cultured into the microorganism enrichment kettle, adding the culture gas or inert gas into the gas storage tank, and adding the nutrient solution into the nutrient bottle;

[0051] S2: Control the real-time temperature of the water bath; control the opening of the air inlet valve, adjust the real-time pressure of the culture gas or inert gas output by the gas injection unit, and input it into the microorganism enrichment kettle; control the opening of the liquid inlet valve, select the nutrient solution, control the opening of the corresponding liquid outlet valve and the second liquid injection pump, input it into the microorganism enrichment kettle, and form a microbial bacterial liquid;

[0052] S3: Controlling the magnetic stirrer to stir the microbial solution, and after a first preset time, controlling the injection pump to pump the microbial solution into the first filter to filter out large particles of impurities, obtain a first-stage filtered microbial solution, and enter the separation chamber of the microbial separator;

[0053] S4: Control the dilution liquid storage bin to open, and the dilution liquid enters the separation bin to dilute the primary filtered microbial solution according to a preset ratio to obtain a diluted microbial solution; control the magnet to open, and the magnetic beads drive the diluted microbial solution in the separation bin to rotate, to obtain a separated diluted microbial solution;

[0054] S5: the separated diluted microbial solution enters the second filter to filter out sediments to obtain a secondary filtered microbial solution;

[0055] S6: The microbial liquid after secondary filtration enters the first separation filter, and the microbial liquid that has not passed through the first filter enters the first input end of the multi-channel single cell sorter through the first output end of the first separation filter; the microbial liquid that has passed through the first filter enters the second filter through the second output end of the first separation filter, and the microbial liquid that has not passed through the second filter enters the second input end of the multi-channel single cell sorter through the first output end of the second separation filter, and the microbial liquid that has passed through the second filter enters the third input end of the multi-channel single cell sorter through the second output end of the second separation filter;

[0056] S7: Observe the state and type of single cells entering each input port through the window on the multi-channel single cell sorter to determine whether the target microbial cells are obtained; if not, execute step S8; otherwise, execute step S9;

[0057] S8: Control the filtration pump to start, extract and discharge the microbial liquid in the separation chamber, and repeat steps S3-S7;

[0058] S9: Control the output end of the multi-channel single cell sorter corresponding to the target microbial cells to open, turn the handle to make the culture cup correspond to the first through hole, and the target microbial cells enter the culture cup for culture; after the second preset time, turn the handle to make the culture cup correspond to the second through hole, control the first injection pump to start, and make the fixative enter the culture cup, so as to complete the fixation of the cultured target microbial cells and obtain a fixed sample;

[0059] S10: transferring the fixed sample to an external multi-omics unit for cell characteristic detection.

[0060] Preferably, the external multi-omics unit comprises an automated DNA extraction instrument and a single-cell sequencer.

[0061] The automatic DNA extraction instrument and single-cell sequencer can obtain the genetic characteristics of a single microorganism, thereby improving the detection efficiency and genome cognition of the isolated single microorganism.

[0062] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0063] The present invention sets a microorganism enrichment unit for microorganism enrichment, uses a multi-sequence nutrient solution supply unit to provide the microorganism enrichment unit with the nutrient solution required for microorganism culture, uses a gas injection unit to provide the microorganism enrichment unit with the culture gas or inert gas required for microorganism culture, and makes the pressure in the microorganism enrichment unit consistent with the pressure value of the microorganism in the ocean situ, thereby ensuring the effectiveness of enrichment culture; after a period of enrichment culture, the microorganism liquid is pumped into a multi-stage separation unit to complete the multi-stage separation of cells of different sizes of the microorganism liquid to obtain target microorganism cells; finally, the target microorganism cells are transferred to a monitoring culture unit for culture and fixation of the target microorganism, and an external multi-omics unit is used to detect cell characteristics. The present invention can continuously separate marine microorganisms of different sizes and then perform characteristic detection in an ocean situ high-pressure environment, is simple to operate, improves separation efficiency, and has accurate detection results that fit the actual environment. BRIEF DESCRIPTION OF THE DRAWINGS

[0064] Figure 1 This is a schematic diagram of the structure of a continuous separation device for marine microorganisms under high pressure environment described in Example 1;

[0065] Figure 2 This is a schematic diagram of the structure of a continuous separation device for marine microorganisms under high pressure environment described in Example 2;

[0066] Figure 3 It is a schematic diagram of the structure of the microbial separator described in Example 2;

[0067] Figure 4This is a schematic diagram of the distribution of the culture cups described in Example 2 on the support frame;

[0068] Figure 5 This is a flow chart of a method for continuous separation of marine microorganisms under high pressure environment described in Example 3;

[0069] In the figure, 1-microorganism enrichment unit, 2-multi-stage separation unit, 3-monitoring culture unit, 4-gas injection unit, 5-multi-sequence nutrient solution supply unit, 6-central control unit, 11-microorganism enrichment kettle, 12-magnetic stirrer, 13-vent valve, 14-vent valve, 15-pressure sensor, 16-temperature sensor, 17-water bath, 18-intake valve, 21-first filter, 22-microorganism separator, 23-second filter, 24-first separation filter, 25-first Second separation filter, 26-multi-channel single cell sorter, 31-culture chamber, 32-support frame, 33-culture cup, 34-handle, 35-environmental indicator sensor, 36-first injection pump, 37-fixed liquid container, 41-boosting pump, 42-air compressor, 43-gas storage tank, 44-pressure regulating valve, 51-nutrient bottle, 52-liquid outlet valve, 53-second injection pump, 221-separation chamber, 222-dilution liquid storage chamber, 223-suction pump, 224-magnetic device, 225-magnetic beads. DETAILED DESCRIPTION

[0070] The drawings are for illustrative purposes only and should not be construed as limiting the present patent;

[0071] In order to better illustrate the present embodiment, some parts in the drawings may be omitted, enlarged or reduced, and do not represent the size of the actual product;

[0072] It is understandable to those skilled in the art that some well-known structures and their descriptions may be omitted in the drawings.

[0073] The technical solution of the present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0074] Example 1

[0075] This embodiment discloses a continuous separation device for marine microorganisms under high pressure environment, such as Figure 1 As shown, it includes: a microorganism enrichment unit 1, a gas injection unit 4, a multi-sequence nutrient solution supply unit 5, a multi-stage separation unit 2, a monitoring and cultivation unit 3 and a central control unit 6;

[0076] The air inlet end of the microorganism enrichment unit 1 is connected to the air outlet end of the gas injection unit 4, and the liquid inlet end of the microorganism enrichment unit 1 is connected to the liquid outlet end of the multi-sequence nutrient solution supply unit 5;

[0077] The input end of the multi-stage separation unit 2 is connected to the liquid outlet end of the microorganism enrichment unit 1, and the output end of the multi-stage separation unit 2 is connected to the input end of the monitoring and cultivation unit 3;

[0078] The data output ends of the microorganism enrichment unit 1 and the monitoring and cultivation unit 3 are connected to the data input end of the central control unit 6, and the control ends of the microorganism enrichment unit 1, the gas injection unit 4, the multi-sequence nutrient solution supply unit 5, the multi-stage separation unit 2, and the monitoring and cultivation unit 3 are connected to the control end of the central control unit 6.

[0079] In the specific implementation process, the present embodiment sets a microorganism enrichment unit 1 for microorganism enrichment, uses a multi-sequence nutrient solution supply unit 5 to provide the microorganism enrichment unit 1 with the nutrient solution required for microorganism culture, uses a gas injection unit 4 to provide the microorganism enrichment unit 1 with the culture gas or inert gas required for microorganism culture, and makes the pressure in the microorganism enrichment unit 1 consistent with the pressure value of the microorganism in situ in the ocean, thereby ensuring the effectiveness of the enrichment culture; after a period of enrichment culture, the microbial liquid is pumped into the multi-stage separation unit 2 to complete the multi-stage separation of cells of different sizes in the microorganism liquid to obtain target microbial cells; finally, the target microbial cells are transferred to the monitoring culture unit 3 for culture and fixation of the target microorganism, and the cell characteristics are detected using an external multi-omics unit.

[0080] Example 2

[0081] The present invention discloses a continuous separation device for marine microorganisms under high pressure environment, comprising: a microorganism enrichment unit 1, a gas injection unit 4, a multi-sequence nutrient solution supply unit 5, a multi-stage separation unit 2, a monitoring and cultivation unit 3 and a central control unit 6;

[0082] The air inlet end of the microorganism enrichment unit 1 is connected to the air outlet end of the gas injection unit 4, and the liquid inlet end of the microorganism enrichment unit 1 is connected to the liquid outlet end of the multi-sequence nutrient solution supply unit 5;

[0083] The input end of the multi-stage separation unit 2 is connected to the liquid outlet end of the microorganism enrichment unit 1, and the output end of the multi-stage separation unit 2 is connected to the input end of the monitoring and cultivation unit 3;

[0084] The microorganism enrichment unit 1 includes a microorganism enrichment kettle 11, a magnetic stirrer 12, a vent valve 13, a liquid inlet valve 14, a pressure sensor 15, a temperature sensor 16, a water bath 17 and an air inlet valve 18;

[0085] The side wall of the microorganism enrichment kettle 11 is provided with a liquid outlet, and the liquid outlet serves as the liquid outlet of the microorganism enrichment unit 1 and is connected to the input end of the first filter 21;

[0086] The microorganism enrichment kettle 11 is arranged in a water bath 17, a magnetic stirrer 12 is arranged on the lower surface of the microorganism enrichment kettle 11, and the control end of the water bath 17 and the magnetic stirrer 12 is connected to the control end of the central control unit 6;

[0087] One end of the vent valve 13 is connected to the upper surface of the microorganism enrichment kettle 11, and the other end of the vent valve 13 is suspended in the air. The control end of the vent valve 13 is connected to the control end of the central control unit 6;

[0088] One end of the air inlet valve 18 is connected to the upper surface of the microorganism enrichment kettle 11, and the other end of the air inlet valve 18 is connected to the gas outlet end of the gas injection unit 4 as the air inlet end of the microorganism enrichment unit 1, and the control end of the air inlet valve 18 is connected to the control end of the central control unit 6;

[0089] One end of the liquid inlet valve 14 is connected to the side wall of the microorganism enrichment kettle 11, and the other end of the liquid inlet valve 14 is connected to the liquid outlet of the multi-sequence nutrient solution supply unit 5 as the liquid inlet end of the microorganism enrichment unit 1, and the control end of the liquid inlet valve 14 is connected to the control end of the central control unit 6;

[0090] The temperature sensor 16 and the pressure sensor 15 are both arranged on the upper surface of the microorganism enrichment kettle 11, and the data output ends of the temperature sensor 16 and the pressure sensor 15 are both connected to the data input end of the central control unit 6;

[0091] The upper surface of the microorganism enrichment kettle 11 is detachable, which is convenient for sterilization and placing the substrate to be cultured; a magnetic stirrer 12 is set on the lower surface to enhance the mass transfer process in the microorganism enrichment kettle 11 and increase the energy and nutrient utilization efficiency of the microorganism liquid. The pressure sensor 15 and the temperature sensor 16 are used to monitor the pressure and temperature changes in the microorganism enrichment kettle 11 in real time, the vent valve 13 is used to reduce the real-time pressure in the microorganism enrichment kettle 11, and the water bath 17 is used to maintain the constant temperature state in the microorganism enrichment kettle 11. The air inlet valve 18 and the liquid inlet valve 14 are used to control the entry of culture gas or inert gas and nutrient solution respectively, so that the microorganism liquid is always in the best growth environment.

[0092] The gas injection unit 4 includes a booster pump 41, an air compressor 42, a gas storage tank 43 and a pressure regulating valve 44 connected in sequence;

[0093] The other end of the pressure regulating valve 44 is connected to the other end of the air inlet valve 18 as the outlet end of the gas injection unit 4; the control end of the booster pump 41, the air compressor 42, and the pressure regulating valve 44 are connected to the control end of the central control unit 6;

[0094] The gas storage tank 43 is filled with culture gas or inert gas.

[0095] The multi-sequence nutrient solution supply unit 5 includes a plurality of nutrient bottles 51, a plurality of liquid outlet valves 52 and a second liquid injection pump 53;

[0096] The liquid outlet of each nutrient bottle 51 is connected to one end of a liquid outlet valve 52, and the other end of each liquid outlet valve 52 is connected to one end of a second liquid injection pump 53. The other end of the second liquid injection pump 53 serves as the liquid outlet end of the multi-sequence nutrient solution supply unit 5 and is connected to the other end of the liquid inlet valve 14. The control end of each liquid outlet valve 52 and the control end of the second liquid injection pump 53 are connected to the control end of the central control unit 6.

[0097] Each of the nutrient bottles 51 is provided with a nutrient solution.

[0098] The multi-stage separation unit 2 includes a first filter 21, a microorganism separator 22, a second filter 23, a first separation filter 24, a second separation filter 25 and a multi-channel single cell sorter 26;

[0099] The first filter 21, the microorganism separator 22, the second filter 23, and the first separation filter 24 are connected in sequence; the first output end of the first separation filter 24 is connected to the first input end of the multi-channel single cell sorter 26, the second output end of the first separation filter 24 is connected to the input end of the second separation filter 25, the first output end of the second separation filter 25 is connected to the second input end of the multi-channel single cell sorter 26, and the second output end of the second separation filter 25 is connected to the third input end of the multi-channel single cell sorter 26;

[0100] The input end of the first filter 21 is used as the input end of the multi-stage separation unit 2, and is connected to the liquid outlet of the microorganism enrichment unit 1. The first output end, the second output end, and the third output end of the multi-channel single cell sorter 26 are used as the output ends of the multi-stage separation unit 2, and are connected to the input end of the monitoring culture unit 3.

[0101] The first filter 21 includes a first filter frame and a first filter membrane, and the surface of the first filter frame is covered with the first filter membrane;

[0102] The second filter 23 includes a second filter frame and a second filter membrane, and the surface of the second filter frame is covered with the second filter membrane;

[0103] The filtration diameter of the second filtration membrane is smaller than the filtration diameter of the first filtration membrane;

[0104] The first separation filter 24 includes a first protective shell and a first filter screen, and the first filter screen is arranged inside the first protective shell;

[0105] The second separation filter 25 includes a second protective shell and a second filter screen, and the second filter screen is arranged inside the second protective shell;

[0106] The filtering diameter of the second filter screen is smaller than the filtering diameter of the first filter screen;

[0107] like Figure 3 As shown, the microorganism separator 22 includes a separation chamber 221, a diluent storage chamber 222, a filtration pump 223, a magnet 224 and magnetic beads 225;

[0108] The magnetic beads 225 are arranged inside the separation chamber 221, and the magnet 224 is arranged on the lower surface of the separation chamber 221, corresponding to the position of the magnetic beads 225;

[0109] The water outlet end of the suction filter pump 223 is suspended in the air, the water inlet end of the suction filter pump 223 extends into the interior of the separation chamber 221, and the control end of the suction filter pump 223 is connected to the control end of the central control unit 6;

[0110] The diluent storage bin 222 is provided with diluent, and the diluent storage bin 222 is connected to the upper surface of the separation bin 221 through a pipeline.

[0111] The microbial liquid is pumped into the first filter 21 to filter out large particles of impurities, such as humus, to obtain a first-stage filtered microbial liquid, which enters the microbial separator 22 for dilution and separation. After the first-stage filtered microbial liquid enters the separation chamber 221 in the microbial separator 22, the diluent storage chamber 222 is opened, and the diluent enters the separation chamber 221 to dilute the first-stage filtered microbial liquid according to a preset ratio to obtain a diluted microbial liquid. The diluent is mainly used to dilute the liquid and protect cells from damage, and can provide certain nutrients. The function of the magnet 224 and the magnetic beads 225 is to drive the diluted microbial liquid in the separation chamber 221 to rotate. , the sediment and the cell-bound part are separated to obtain a separated microbial diluted bacterial solution; the separated microbial diluted bacterial solution enters the second filter 23 to filter out sediments larger than the diameter of the microorganisms in the separated microbial diluted bacterial solution to obtain a secondary filtered microbial bacterial solution; the filtration diameter of the first separation filter 24 is larger than the filtration diameter of the second separation filter 25, and the secondary filtered microbial bacterial solution first enters the first separation filter 24 for primary separation, and the microbial bacterial solution that does not pass through enters the first input end of the multi-channel single cell sorter 26 through the first output end of the first separation filter 24; the microbial bacterial solution that passes through the first separation filter 24 The second output end of the filter 24 enters the second filter 23 for secondary separation, the microbial solution that does not pass through enters the second input end of the multi-channel single cell sorter 26 through the first output end of the second separation filter 25, and the microbial solution that passes through enters the third input end of the multi-channel single cell sorter 26 through the second output end of the second separation filter 25, completing the multi-stage separation of cells of different sizes of the microbial solution; the first separation filter 24 and the second separation filter 25 are used to separate cells of different diameters. Since the filtration diameter of the second filter 252 is smaller than the filtration diameter of the first filter 242, the microbial solution filtered at the second level first enters the first separation filter 24 A primary separation is performed, and the cell size of the microbial liquid that has not passed through the first filter 242 is the largest, and it enters the multi-channel single cell sorter 26 through the first output end of the first separation filter 24; the microbial liquid that has passed through a filter enters the second separation filter 25 through the second output end of the first separation filter 24 for secondary separation, and the cell size of the microbial liquid that has not passed through the second filter 252 is the second largest, and it enters the multi-channel single cell sorter 26 through the first output end of the second separation filter 25; the cell size of the microbial liquid that has passed through the second filter 252 is the smallest, and it enters the multi-channel single cell sorter 26 through the second output end of the second separation filter 25.After entering the multi-channel single-cell sorter 26, the state and type of the single cells entering from each input port are observed through the window on the multi-channel single-cell sorter 26 to determine whether the target microbial cells are obtained. If the target microbial cells are not obtained, the primary filtered microbial liquid in the separation chamber 221 is extracted and discharged by the filtration pump 223, and the primary filtered microbial liquid in the microbial separator 22 is updated, and the above steps are repeated until the target microbial cells are obtained.

[0112] The monitoring and culture unit 3 includes a culture chamber 31, a support frame 32, a plurality of culture cups 33, a handle 34, an environmental indicator sensor 35, a first injection pump 36 and a fixed liquid container 37;

[0113] The environmental indicator sensor 35, the support frame 32 and the plurality of culture cups 33 are arranged in the culture chamber 31, and the data output end of the environmental indicator sensor 35 is connected to the data input end of the central control unit 6;

[0114] like Figure 4 As shown, the plurality of culture cups 33 are evenly arranged on the upper surface of the support frame 32 along the circumference; the handle 34 is vertically arranged, one end of which is connected to the lower surface of the support frame 32, and the other end extends from the bottom surface of the culture chamber 31;

[0115] The upper surface of the culture chamber 31 is provided with a first through hole and a second through hole, and the positions of the first through hole and the second through hole correspond to any two culture cups 33;

[0116] The first output end, the second output end, and the third output end of the multi-channel single cell sorter 26 are connected to the first through hole; the output end of the fixed liquid container 37 is connected to one end of the first injection pump 36, and the other end of the first injection pump 36 is connected to the second through hole;

[0117] The environmental indicator sensor 35 includes an environmental pressure sensor, an environmental temperature sensor and a gas concentration detector;

[0118] The culture chamber 31 is cylindrical, the support frame 32 is circular, and a number of culture cups 33 are evenly arranged on the upper surface of the support frame 32 along the circumference. The handle 34 is vertically connected to the lower surface of the support frame 32. The upper surface of the culture chamber 31 is provided with a first through hole and a second through hole, which correspond to any two culture cups 33 respectively; when in use, the output end of the multi-channel single-cell sorter 26 corresponding to the target microbial cells is opened, and enters the culture cup 33 through the first through hole for culture; the handle 34 is turned to make the next culture cup 33 correspond to the first through hole; after a period of culture, the handle 34 is turned to make the culture cup 33 containing the cultured target microbial cells correspond to the second through hole, and the first injection pump 36 is controlled to start, so that the fixative enters the culture cup 33, and the fixation of the cultured target microbial cells is completed to obtain a fixed sample.

[0119] This embodiment can continuously separate marine microorganisms of different sizes in an in-situ high-pressure environment in the ocean and then perform characteristic detection. The operation is simple, the separation efficiency is improved, the detection results are accurate, and they are in line with the actual environment.

[0120] Example 3

[0121] This embodiment provides a method for continuous separation of marine microorganisms under high pressure, which is applied to the continuous separation device described in Embodiment 1 or 2, such as Figure 3 As shown, including:

[0122] S1: sterilizing the microorganism enrichment unit, the multi-stage separation unit and the monitoring culture unit; adding the substrate to be cultured into the microorganism enrichment kettle, adding the culture gas or inert gas into the gas storage tank, and adding the nutrient solution into the nutrient bottle;

[0123] S2: Control the real-time temperature of the water bath; control the opening of the air inlet valve, adjust the real-time pressure of the culture gas or inert gas output by the gas injection unit, and input it into the microorganism enrichment kettle; control the opening of the liquid inlet valve, select the nutrient solution, control the opening of the corresponding liquid outlet valve and the second liquid injection pump, input it into the microorganism enrichment kettle, and form a microbial bacterial liquid;

[0124] S3: Controlling the magnetic stirrer to stir the microbial solution, and after a first preset time, controlling the injection pump to pump the microbial solution into the first filter to filter out large particles of impurities, obtain a first-stage filtered microbial solution, and enter the separation chamber of the microbial separator;

[0125] S4: Control the dilution liquid storage bin to open, and the dilution liquid enters the separation bin to dilute the primary filtered microbial solution according to a preset ratio to obtain a diluted microbial solution; control the magnet to open, and the magnetic beads drive the diluted microbial solution in the separation bin to rotate, to obtain a separated diluted microbial solution;

[0126] S5: the separated diluted microbial solution enters the second filter to filter out sediments to obtain a secondary filtered microbial solution;

[0127] S6: The microbial liquid after secondary filtration enters the first separation filter, and the microbial liquid that has not passed through the first filter enters the first input end of the multi-channel single cell sorter through the first output end of the first separation filter; the microbial liquid that has passed through the first filter enters the second filter through the second output end of the first separation filter, and the microbial liquid that has not passed through the second filter enters the second input end of the multi-channel single cell sorter through the first output end of the second separation filter, and the microbial liquid that has passed through the second filter enters the third input end of the multi-channel single cell sorter through the second output end of the second separation filter;

[0128] S7: Observe the state and type of single cells entering each input port through the window on the multi-channel single cell sorter to determine whether the target microbial cells are obtained; if not, execute step S8; otherwise, execute step S9;

[0129] S8: Control the filtration pump to start, extract and discharge the microbial liquid in the separation chamber, and repeat steps S3-S7;

[0130] S9: Control the output end of the multi-channel single cell sorter corresponding to the target microbial cells to open, turn the handle to make the culture cup correspond to the first through hole, and the target microbial cells enter the culture cup for culture; after the second preset time, turn the handle to make the culture cup correspond to the second through hole, control the first injection pump to start, and make the fixative enter the culture cup, so as to complete the fixation of the cultured target microbial cells and obtain a fixed sample;

[0131] S10: transferring the fixed sample to an external multi-omics unit for cell characteristic detection.

[0132] The external multi-omics unit includes an automated DNA extraction instrument and a single-cell sequencer.

[0133] The automatic DNA extraction instrument and single-cell sequencer can obtain the genetic characteristics of a single microorganism, thereby improving the detection efficiency and genome cognition of the isolated single microorganism.

[0134] The same or similar reference numerals correspond to the same or similar components;

[0135] The terms used in the drawings to describe positional relationships are only used for illustrative purposes and should not be construed as limiting this patent;

[0136] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the embodiments here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A continuous separation device for marine microorganisms under high pressure environment, characterized in that: include: A microorganism enrichment unit (1), a multi-stage separation unit (2), a monitoring and cultivation unit (3), a gas injection unit (4), a multi-sequence nutrient solution supply unit (5) and a central control unit (6); The air inlet end of the microorganism enrichment unit (1) is connected to the air outlet end of the gas injection unit (4), and the liquid inlet end of the microorganism enrichment unit (1) is connected to the liquid outlet end of the multi-sequence nutrient solution supply unit (5); The input end of the multi-stage separation unit (2) is connected to the liquid outlet end of the microorganism enrichment unit (1), and the output end of the multi-stage separation unit (2) is connected to the input end of the monitoring and cultivation unit (3); The data output ends of the microorganism enrichment unit (1) and the monitoring and cultivation unit (3) are connected to the data input end of the central control unit (6), and the control ends of the microorganism enrichment unit (1), the gas injection unit (4), the multi-sequence nutrient solution supply unit (5), the multi-stage separation unit (2), and the monitoring and cultivation unit (3) are connected to the control end of the central control unit (6).

2. The continuous separation device for marine microorganisms under high pressure environment according to claim 1, characterized in that: The multi-stage separation unit (2) comprises a first filter (21), a microorganism separator (22), a second filter (23), a first separation filter (24), a second separation filter (25) and a multi-channel single cell sorter (26); The first filter (21), the microorganism separator (22), the second filter (23), and the first separation filter (24) are connected in sequence; the first output end of the first separation filter (24) is connected to the first input end of the multi-channel single cell sorter (26), the second output end of the first separation filter (24) is connected to the input end of the second separation filter (25), the first output end of the second separation filter (25) is connected to the second input end of the multi-channel single cell sorter (26), and the second output end of the second separation filter (25) is connected to the third input end of the multi-channel single cell sorter (26); The input end of the first filter (21) serves as the input end of the multi-stage separation unit (2) and is connected to the liquid outlet end of the microorganism enrichment unit (1); the first output end, the second output end, and the third output end of the multi-channel single cell sorter (26) all serve as the output ends of the multi-stage separation unit (2) and are connected to the input end of the monitoring culture unit (3).

3. The continuous separation device for marine microorganisms under high pressure environment according to claim 2, characterized in that: The first filter (21) comprises a first filter frame and a first filter membrane, and the surface of the first filter frame is covered with the first filter membrane; The second filter (23) comprises a second filter frame and a second filter membrane, and the surface of the second filter frame is covered with the second filter membrane; The filtration diameter of the second filtration membrane is smaller than the filtration diameter of the first filtration membrane.

4. The continuous separation device for marine microorganisms under high pressure environment according to claim 2, characterized in that: The first separation filter (24) comprises a first protective shell and a first filter screen, wherein the first filter screen is arranged inside the first protective shell; The second separation filter (25) comprises a second protective shell and a second filter screen, wherein the second filter screen is arranged inside the second protective shell; The filtering diameter of the second filter net is smaller than the filtering diameter of the first filter net.

5. The continuous separation device for marine microorganisms under high pressure environment according to claim 2, characterized in that: The microorganism separator (22) comprises a separation chamber (221), a diluent storage chamber (222), a filtration pump (223), a magnetizer (224) and magnetic beads (225); The magnetic beads (225) are arranged inside the separation chamber (221), and the magnet (224) is arranged on the lower surface of the separation chamber (221) corresponding to the position of the magnetic beads (225); The water outlet end of the suction filter pump (223) is suspended in the air, the water inlet end of the suction filter pump (223) extends into the interior of the separation chamber (221), and the control end of the suction filter pump (223) is connected to the control end of the central control unit (6); The diluent storage bin (222) contains diluent, and the diluent storage bin (222) is connected to the upper surface of the separation bin (221) via a pipeline.

6. The continuous separation device for marine microorganisms under high pressure environment according to any one of claims 1 to 5, characterized in that: The monitoring culture unit (3) comprises a culture chamber (31), a support frame (32), a plurality of culture cups (33), a handle (34), an environmental indicator sensor (35), a first injection pump (36) and a fixed liquid container (37); The environmental indicator sensor (35), the support frame (32) and the plurality of culture cups (33) are arranged in the culture chamber (31), and the data output end of the environmental indicator sensor (35) is connected to the data input end of the central control unit (6); The plurality of culture cups (33) are evenly arranged on the upper surface of the support frame (32) along the circumference; the handle (34) is vertically arranged, one end of which is connected to the lower surface of the support frame (32) and the other end of which extends from the bottom surface of the culture chamber (31); The upper surface of the culture chamber (31) is provided with a first through hole and a second through hole, and the positions of the first through hole and the second through hole correspond to any two culture cups (33); The first output end, the second output end, and the third output end of the multi-channel single cell sorter (26) are connected to the first through hole; the output end of the fixed liquid container (37) is connected to one end of the first injection pump (36), and the other end of the first injection pump (36) is connected to the second through hole.

7. The continuous separation device for marine microorganisms under high pressure environment according to claim 6, characterized in that: The microorganism enrichment unit (1) comprises a microorganism enrichment kettle (11), a magnetic stirrer (12), a vent valve (13), a liquid inlet valve (14), a pressure sensor (15), a temperature sensor (16), a water bath (17) and an air inlet valve (18); The side wall of the microorganism enrichment kettle (11) is provided with a liquid outlet, and the liquid outlet serves as the liquid outlet of the microorganism enrichment unit (1) and is connected to the input end of the first filter (21); The microorganism enrichment kettle (11) is arranged in a water bath (17), a magnetic stirrer (12) is arranged on the lower surface of the microorganism enrichment kettle (11), and the control ends of the water bath (17) and the magnetic stirrer (12) are connected to the control end of the central control unit (6); One end of the vent valve (13) is connected to the upper surface of the microorganism enrichment kettle (11), the other end of the vent valve (13) is suspended in the air, and the control end of the vent valve (13) is connected to the control end of the central control unit (6); One end of the air inlet valve (18) is connected to the upper surface of the microorganism enrichment kettle (11), the other end of the air inlet valve (18) is connected to the gas outlet end of the gas injection unit (4) as the air inlet end of the microorganism enrichment unit (1), and the control end of the air inlet valve (18) is connected to the control end of the central control unit (6); One end of the liquid inlet valve (14) is connected to the side wall of the microorganism enrichment kettle (11), the other end of the liquid inlet valve (14) is connected to the liquid outlet of the multi-sequence nutrient solution supply unit (5) as the liquid inlet end of the microorganism enrichment unit (1), and the control end of the liquid inlet valve (14) is connected to the control end of the central control unit (6); The temperature sensor (16) and the pressure sensor (15) are both arranged on the upper surface of the microorganism enrichment kettle (11), and the data output ends of the temperature sensor (16) and the pressure sensor (15) are both connected to the data input end of the central control unit (6).

8. The continuous separation device for marine microorganisms under high pressure environment according to claim 6, characterized in that: The gas injection unit (4) comprises a booster pump (41), an air compressor (42), a gas storage tank (43) and a pressure regulating valve (44) which are connected in sequence; The other end of the pressure regulating valve (44) is connected to the other end of the air inlet valve (18) as the air outlet end of the gas injection unit (4); the control ends of the booster pump (41), the air compressor (42) and the pressure regulating valve (44) are connected to the control end of the central control unit (6); The gas storage tank (43) is filled with culture gas or inert gas.

9. The continuous separation device for marine microorganisms under high pressure environment according to claim 6, characterized in that: The multi-sequence nutrient solution supply unit (5) comprises a plurality of nutrient bottles (51), a plurality of liquid outlet valves (52) and a second liquid injection pump (53); The liquid outlet of each of the nutrient bottles (51) is connected to one end of a corresponding liquid outlet valve (52); the other end of each of the liquid outlet valves (52) is connected to one end of a second liquid injection pump (53); the other end of the second liquid injection pump (53) serves as the liquid outlet of the multi-sequence nutrient solution supply unit (5) and is connected to the other end of the liquid inlet valve (14); the control end of each liquid outlet valve (52) and the control end of the second liquid injection pump (53) are connected to the control end of the central control unit (6); A nutrient solution is arranged in each of the nutrient bottles (51).

10. A method for continuous separation of marine microorganisms under high pressure, applied to the continuous separation device according to any one of claims 1 to 9, characterized in that: include: S1: sterilizing the microorganism enrichment unit, the multi-stage separation unit and the monitoring culture unit; adding the substrate to be cultured into the microorganism enrichment kettle, adding the culture gas or inert gas into the gas storage tank, and adding the nutrient solution into the nutrient bottle; S2: Control the real-time temperature of the water bath; control the opening of the air inlet valve, adjust the real-time pressure of the culture gas or inert gas output by the gas injection unit, and input it into the microorganism enrichment kettle; control the opening of the liquid inlet valve, select the nutrient solution, control the opening of the corresponding liquid outlet valve and the second liquid injection pump, input it into the microorganism enrichment kettle, and form a microbial bacterial liquid; S3: Controlling the magnetic stirrer to stir the microbial solution, and after a first preset time, controlling the injection pump to pump the microbial solution into the first filter to filter out large particles of impurities, obtain a first-stage filtered microbial solution, and enter the separation chamber of the microbial separator; S4: Control the dilution liquid storage bin to open, and the dilution liquid enters the separation bin to dilute the primary filtered microbial solution according to a preset ratio to obtain a diluted microbial solution; control the magnet to open, and the magnetic beads drive the diluted microbial solution in the separation bin to rotate, to obtain a separated diluted microbial solution; S5: the separated diluted microbial solution enters the second filter to filter out sediments to obtain a secondary filtered microbial solution; S6: The microbial liquid after secondary filtration enters the first separation filter, and the microbial liquid that has not passed through the first filter enters the first input end of the multi-channel single cell sorter through the first output end of the first separation filter; the microbial liquid that has passed through the first filter enters the second filter through the second output end of the first separation filter, and the microbial liquid that has not passed through the second filter enters the second input end of the multi-channel single cell sorter through the first output end of the second separation filter, and the microbial liquid that has passed through the second filter enters the third input end of the multi-channel single cell sorter through the second output end of the second separation filter; S7: Observe the state and type of single cells entering from each input port through the window on the multi-channel single cell sorter to determine whether the target microbial cells are obtained; If not obtained, execute step S8, otherwise, execute step S9; S8: Control the filtration pump to start, extract and discharge the microbial liquid in the separation chamber, and repeat steps S3-S7; S9: Control the output end of the multi-channel single cell sorter corresponding to the target microbial cells to open, turn the handle to make the culture cup correspond to the first through hole, and the target microbial cells enter the culture cup for culture; after the second preset time, turn the handle to make the culture cup correspond to the second through hole, control the first injection pump to start, and make the fixative enter the culture cup, so as to complete the fixation of the cultured target microbial cells and obtain a fixed sample; S10: transferring the fixed sample to an external multi-omics unit for cell characteristic detection.

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