Micro-droplet culture device

Abstract

The utility model provides a micro-droplet culture device, belongs to the field of droplet microfluidics, and aims to solve the problem that an existing micro-droplet culture device cannot effectively avoid coagulation and huddling phenomena of strains, the micro-droplet culture device comprises a device shell, a first mounting rack and a second mounting rack are fixed in the device shell, and a first pinch valve, a second pinch valve, a third pinch valve, a fourth pinch valve, a fifth pinch valve, a sixth pinch valve, a seventh pinch valve, an eighth pinch valve, a ninth pinch valve and a tenth pinch valve are fixed on the second mounting frame. The first pinch valve to the ninth pinch valve are sequentially opened or closed, driving force is provided in cooperation with the oil pump injection pump and the liquid pump injection pump, liquid drops can reciprocate in the positive and negative directions in the culture process, it can be guaranteed that microorganisms can be evenly distributed in the liquid drops in the liquid drop culture process, and the liquid drop culture efficiency is improved. Therefore, the influence on subsequent liquid drop detection work caused by liquid drop coagulation can be effectively avoided.

Description

TECHNICAL FIELD

[0001] The utility model relates to the field of droplet microfluidics, in particular to a microdroplet culture device. BACKGROUND

[0002] The development of the microbial industry is closely related to our daily life, and affects our life, health and environment. The microbial industry has occupied a pivotal position in the food, medicine, energy and environmental protection industries. Microbial breeding is a very important part of the microbial industry, including strain library construction, screening and culture.

[0003] In recent years, with the development of microfluidic technology, microbial breeding has entered a new era. Microfluidic technology has been applied to the microbial breeding industry and has achieved good results in microbial cell culture and screening. For example, a microliter-scale single cell droplet generation and culture method and device are disclosed in patent CN115521882A. However, in the process of preparing droplets, the water phase can be squeezed and divided into multiple initial droplets by the oil phase in the two oil phase flow channels. According to the prior art, the droplets formed by the oil and water phases are prone to contact and fusion during movement. The surfactant can only maintain the stability of nanoliter-scale and below droplets, but cannot prevent the mutual fusion of microliter-scale droplets. Therefore, the culture in a closed container cannot meet the oxygen demand. Moreover, the existing culture methods are static culture in the pipeline, which can only be applied to environmental biological samples, samples with no special requirements for the consistency of detection results of the same batch of droplets, and strains with good dispersion and not easy to aggregate. However, for strains that are easy to aggregate and clump, the detection of droplets may be affected by the aggregation of bacterial bodies, resulting in inconsistent detection results of the same batch of droplets, which may not meet the requirements. Therefore, we have improved the microdroplet culture device. SUMMARY

[0004] The purpose of the utility model is to solve the problem that the existing microdroplet culture device cannot prevent the mutual fusion of microliter-scale droplets and cannot effectively prevent the aggregation and clumping of strains.

[0005] To achieve the above purpose, the utility model provides the following technical scheme:

[0006] The microdroplet culture device is used to improve the above problems.

[0007] The application is as follows:

[0008] The device shell is internally provided with an oil pump injection pump, a liquid pump injection pump and a gas pump injection pump, the first mounting frame and the second mounting frame are fixed in the device shell, the first clamp pipe valve, the second clamp pipe valve, the third clamp pipe valve, the fourth clamp pipe valve, the fifth clamp pipe valve, the sixth clamp pipe valve, the seventh clamp pipe valve, the eighth clamp pipe valve, the ninth clamp pipe valve and the tenth clamp pipe valve are fixed on the second mounting frame, the tail valve is fixed on the first mounting frame, the bubble removal oil bottle, the sample injection bottle and the waste liquid outlet are mounted on the first mounting frame, the drop collection module and the clamp jaw are mounted on the inner bottom end face of the device shell, and the hole plate placement position is fixed on the inner bottom end face of the device shell.

[0009] As a preferred technical scheme of the present application, the oil pump injection pump, the liquid pump injection pump and the gas pump injection pump are uniformly distributed in the device shell.

[0010] As a preferred technical scheme of the present application, the first mounting frame is provided with a culture bin, the heater is mounted in the culture bin, the fan is mounted on the top of the heater, the temperature sensor is fixed in the culture bin, and the first drop disc, the second drop disc, the third drop disc and the fourth drop disc are mounted in the culture bin.

[0011] As a preferred technical scheme of the present application, the first clamp pipe valve, the second clamp pipe valve, the third clamp pipe valve, the fourth clamp pipe valve, the fifth clamp pipe valve, the sixth clamp pipe valve, the seventh clamp pipe valve, the eighth clamp pipe valve, the ninth clamp pipe valve and the tenth clamp pipe valve are of the same structure size.

[0012] As a preferred technical scheme of the present application, the first clamp pipe valve, the second clamp pipe valve, the third clamp pipe valve, the fourth clamp pipe valve, the fifth clamp pipe valve, the sixth clamp pipe valve, the seventh clamp pipe valve, the eighth clamp pipe valve, the ninth clamp pipe valve and the tenth clamp pipe valve are uniformly distributed on the second mounting frame.

[0013] As a preferred technical scheme of the present application, the heater, the fan and the temperature sensor are arranged at the inner center part of the culture bin.

[0014] Compared with the prior art, the present application has the following beneficial effects:

[0015] In the scheme of the present application:

[0016] 1. The gas pump injection pump is arranged, the gas phase is generated by the gas pump injection pump, and the gas phase serves as the intermediate phase of the oil phase and the liquid phase, each liquid drop is wrapped by the oil phase on both sides, and each liquid drop wrapped by the oil phase is spaced apart by the gas phase; since each liquid drop is spaced apart by the gas phase in the middle, fusion between the liquid drops can be avoided, so that the stability of the liquid drops is increased.

[0017] 2. By sequentially opening or closing the first to ninth pinch valves, cooperating with the oil pump injection pump and the liquid pump injection pump to provide driving force, the droplets can realize reciprocating motion in positive and reverse directions during cultivation, so that the microorganisms can be uniformly distributed in the droplets during droplet cultivation, thereby effectively avoiding the influence of droplet aggregation on subsequent droplet detection work. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 The overall structure schematic diagram of the micro-droplet cultivation device provided in the application is shown in the figure.

[0019] Figure 2 The first mounting bracket and the second mounting bracket connection structure schematic diagram of the micro-droplet cultivation device provided in the application is shown in the figure.

[0020] Figure 3 The first pinch valve structure schematic diagram of the micro-droplet cultivation device provided in the application is shown in the figure.

[0021] Figure 4 The Figure 3 The enlarged structure schematic diagram of the micro-droplet cultivation device provided in the application is shown in the figure.

[0022] In the figure, 1 is a device shell, 2 is an oil pump injection pump, 3 is a liquid pump injection pump, 4 is a gas pump injection pump, 5 is a fan, 6 is a first mounting bracket, 7 is a second mounting bracket, 8 is a first pinch valve, 9 is a second pinch valve, 10 is a third pinch valve, 11 is a fourth pinch valve, 12 is a fifth pinch valve, 13 is a sixth pinch valve, 14 is a seventh pinch valve, 15 is an eighth pinch valve, 16 is a ninth pinch valve, 17 is a tenth pinch valve, 18 is a tail valve, 19 is a bubble removal oil bottle, 20 is a sample feeding bottle, 21 is a waste liquid outlet, 22 is a first droplet disc, 23 is a second droplet disc, 24 is a third droplet disc, 25 is a fourth droplet disc, 26 is a temperature sensor, 27 is a droplet collection module, 28 is a clamping jaw, 29 is a hole plate placement position, 30 is a cultivation bin, and 31 is a heater. DETAILED DESCRIPTION

[0023] In order to make the purpose, technical scheme and advantages of the embodiments of the present application more clear, the technical scheme in the embodiments of the present application will be described clearly and completely below in conjunction with the drawings. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments.

[0024] Therefore, the following detailed description of the embodiments of the present application is not intended to limit the scope of the claimed application, but only represents some embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0025] It should be noted that, unless otherwise specified, the embodiments and features and technical solutions in the present invention can be combined with each other.

[0026] It should be noted that similar labels and letters in the following figures indicate similar items. Therefore, once an item is defined in one figure, it does not need to be further defined and explained in subsequent figures.

[0027] In the description of this utility model, it should be noted that the terms "upper," "lower," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, or the orientation or positional relationship commonly used when the product is in use, or the orientation or positional relationship commonly understood by those skilled in the art. These terms are only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation on this utility model. In addition, the terms "first," "second," etc., are only used to distinguish descriptions and should not be construed as indicating or implying relative importance.

[0028] Example 1:

[0029] like Figures 1-3 As shown, this embodiment proposes a microdroplet culture device, including a device housing 1. An oil pump injection pump 2, a liquid pump injection pump 3, and a gas pump injection pump 4 are installed inside the device housing 1. A first mounting bracket 6 and a second mounting bracket 7 are fixed inside the device housing 1. A first clamp valve 8, a second clamp valve 9, a third clamp valve 10, a fourth clamp valve 11, a fifth clamp valve 12, a sixth clamp valve 13, a seventh clamp valve 14, an eighth clamp valve 15, a ninth clamp valve 16, and a tenth clamp valve 17 are fixed on the second mounting bracket 7. A tail valve 18 is fixed on the first mounting bracket 6. An air bubble removal oil bottle 19, a sample inlet bottle 20, and a waste liquid outlet 21 are installed on the first mounting bracket 6. A droplet collection module 27 and a clamp 28 are installed on the inner bottom surface of the device housing 1. A perforated plate placement position 29 is fixed on the inner bottom surface of the device housing 1.

[0030] Example 2:

[0031] The solution in Example 1 will be further described below with reference to its specific working method.

[0032] like Figure 3As shown, in a preferred embodiment, based on the above method, the oil pump injection pump 2, the liquid pump injection pump 3, and the air pump injection pump 4 are evenly distributed inside the device housing 1. The air pump injection pump 4 generates a gas phase, which serves as an intermediate phase between the oil phase and the liquid phase. Each droplet is surrounded by the oil phase on both sides, and each droplet surrounded by the oil phase is separated by the gas phase. Since each droplet is separated by the gas phase, the merging of droplets can be avoided, thereby increasing the stability of the droplets.

[0033] like Figure 4 As shown, in a preferred embodiment, based on the above method, a culture chamber 30 is further installed on the first mounting frame 6. A heater 31 is installed inside the culture chamber 30, and a fan 5 is installed on the top of the heater 31. A first drip tray 22, a second drip tray 23, a third drip tray 24, and a fourth drip tray 25 are installed inside the culture chamber 30. A temperature sensor 26 is fixed inside the culture chamber 30. The heater 31, the fan 5, and the temperature sensor 26 are all located in the center of the culture chamber 30. During the movement of the droplets, the heater 31 and the fan 5 are turned on and off respectively to control the heating and heat dissipation inside the culture device. The temperature sensor 26 provides real-time temperature information, which can determine the working status of the heater 31 and the fan 5.

[0034] like Figure 2 and Figure 3 As shown, in a preferred embodiment, based on the above method, the first clamp valve 8, the second clamp valve 9, the third clamp valve 10, the fourth clamp valve 11, the fifth clamp valve 12, the sixth clamp valve 13, the seventh clamp valve 14, the eighth clamp valve 15, the ninth clamp valve 16, the tenth clamp valve 17, and the tail valve 18 have the same structural size. The first clamp valve 8, the second clamp valve 9, the third clamp valve 10, the fourth clamp valve 11, the fifth clamp valve 12, the sixth clamp valve 13, the seventh clamp valve 14, the eighth clamp valve 15, the ninth clamp valve 16, the tenth clamp valve 17, and the tail valve 18 have the same structural size. Seventh clamp valve 14, eighth clamp valve 15, ninth clamp valve 16, and tenth clamp valve 17 are evenly distributed on the second mounting bracket 7. By sequentially opening or closing the first clamp valve 8 to the ninth clamp valve 16, and with the driving force provided by the oil pump injection pump 2 and the liquid pump injection pump 3, the droplets can achieve reciprocating motion in both directions during the cultivation process. This ensures that the microorganisms are evenly distributed within the droplets during the cultivation process, thereby effectively preventing droplet aggregation that could affect subsequent droplet detection.

[0035] Specifically, when using this microdroplet culture device: First, the oil pump injection pump 2, liquid pump injection pump 3, and air pump injection pump 4 are all connected to different ports of the same cross-shaped connector via pipelines. The last port of the cross-shaped connector is connected to the first port of the T-shaped connector via a pipeline. The second port of the T-shaped connector is connected to another pipeline, and this pipeline is connected to the first waste liquid bottle via the tenth clamp valve 17. The ends of the pipelines connected to the first drop plate 22, the second drop plate 23, the third drop plate 24, and the fourth drop plate 25 converge into the detection pipeline via pipelines connected to the six-way connector. The detection pipeline is connected to the waste liquid pipe via the waste liquid scoop, and the waste liquid pipe is connected to the second waste liquid bottle.

[0036] The third port of the T-connector is connected in parallel to the first drip tray 22, the second drip tray 23, the third drip tray 24 and the fourth drip tray 25 via flexible tubing; and the flexible tubing connected to the first drip tray 22, the second drip tray 23, the third drip tray 24 and the fourth drip tray 25 is respectively inserted into the first clamp valve 8, the second clamp valve 9, the third clamp valve 10 and the fourth clamp valve 11.

[0037] Furthermore, the other ends of the first drip tray 22, the second drip tray 23, the third drip tray 24, and the fourth drip tray 25 are also connected to the second waste bottle in parallel via flexible tubes. The flexible tubes connected to the other ends of the first drip tray 22, the second drip tray 23, the third drip tray 24, and the fourth drip tray 25 are respectively inserted into the sixth clamp valve 13, the seventh clamp valve 14, the eighth clamp valve 15, and the ninth clamp valve 16; the tenth clamp valve 17 is inserted into the pipeline connected to the first waste bottle, and the tail valve 18 is inserted into the pipeline connected to the second waste bottle.

[0038] The first port of the cross-joint is used to introduce an oil phase that is immiscible with the sample, the second port is used to introduce the sample, and the third port is used to introduce gas. At this time, water-in-oil droplets are formed at the cross-joint with the sample. The droplets are separated by air bubbles. Before the droplets are formed, a portion of the oil phase is pushed into the pipeline. During forward movement, the first clamp valve 8 and the sixth clamp valve 13 are opened, the tail valve 18 is opened, and the other valves are closed. At this time, the passage of the first droplet tray 22 is opened, and the oil pump injection pump 2 pushes the droplets a certain distance from the cross-joint.

[0039] The first pinch valve 8 and the sixth pinch valve 13 are closed, and the tail valve 18 is closed; the second pinch valve 9 and the seventh pinch valve 14 are open, the tail valve 18 is open, and the other valves are closed. At this time, the passage of the second drip tray 23 is open, and the oil pump injection pump 2 pushes the droplet a certain distance from the cross joint; the second pinch valve 9 and the seventh pinch valve 14 are closed, and the tail valve 18 is closed; the third pinch valve 10 and the eighth pinch valve 15 are open, the tail valve 18 is open, and the other valves are closed. The passage of the third drip tray 24 is open, and the oil pump injection pump 2 pushes the droplet a certain distance from the cross joint; the third pinch valve 10 and the eighth pinch valve 15 are closed, and the tail valve 18 is closed; the fourth pinch valve 11 and the ninth pinch valve 16 are open, the tail valve 18 is open, and the other valves are closed. The passage of the fourth drip tray 25 is open, and the oil pump injection pump pushes the droplet a certain distance from the cross joint; the fourth pinch valve 11 and the ninth pinch valve 16 are closed, and the tail valve 18 is closed. The forward movement is completed.

[0040] During reverse movement, the first pinch valve 8 and the sixth pinch valve 13 open, the tenth pinch valve 17 opens, and the other valves close. The passage of the first drip tray 22 opens, and the liquid pump 3 pushes the droplet through the six-way connector. The first pinch valve 8 and the sixth pinch valve 13 close, and the tenth pinch valve 17 closes. The second pinch valve 9 and the seventh pinch valve 14 open, the tenth pinch valve 17 opens, and the other valves close. The passage of the second drip tray 23 opens, and the liquid pump 3 pushes the droplet through the six-way connector. The second pinch valve 9 and the seventh pinch valve 14 close, and the tenth pinch valve 17 closes. The third... Pinch valve 10 and eighth pinch valve 15 open, tenth pinch valve 17 opens, other valves close, the passage of the third drip tray 24 opens, and the liquid pump 3 pushes the droplet through the six-way connector; third pinch valve 10 and eighth pinch valve 15 close, and tenth pinch valve 17 closes; fourth pinch valve 11 and ninth pinch valve 16 open, tenth pinch valve 17 opens, other valves close, the passage of the fourth drip tray 25 opens, and the liquid pump 3 pushes the droplet through the six-way connector; fourth pinch valve 11 and ninth pinch valve 16 close, and tenth pinch valve 17 closes, completing the reverse movement.

[0041] During the droplet movement, the heater 31 and fan 5 are turned on and off respectively to control the heating and heat dissipation in the culture device. The temperature sensor 26 provides real-time temperature information to determine the working status of the heater 31 and fan 5. In addition, aerobic bacteria will consume bubbles during the reciprocating culture process, causing the bubbles to become smaller. The use of intermittent reciprocating culture can reduce bubble consumption.

[0042] The above embodiments are only used to illustrate the present utility model and are not intended to limit the technical solutions described in the present utility model. Although the present utility model has been described in detail with reference to the above embodiments, the present utility model is not limited to the specific embodiments described above. Therefore, any modifications or equivalent substitutions to the present utility model, as well as all technical solutions and improvements that do not depart from the spirit and scope of practicality, are covered within the scope of the claims of the present utility model.

Claims

1. A microdroplet cultivation device comprising a device housing (1), characterized in that, The device shell (1) is provided with an oil pump injection pump (2), a liquid pump injection pump (3) and a gas pump injection pump (4), the device shell (1) is fixedly provided with a first mounting frame (6) and a second mounting frame (7), the second mounting frame (7) is fixedly provided with a first pinch valve (8), a second pinch valve (9), a third pinch valve (10), a fourth pinch valve (11), a fifth pinch valve (12), a sixth pinch valve (13), a seventh pinch valve (14), an eighth pinch valve (15), a ninth pinch valve (16) and a tenth pinch valve (17), the first mounting frame (6) is fixedly provided with a tail valve (18), the first mounting frame (6) is provided with a bubble removal oil bottle (19), a sample injection bottle (20) and a waste liquid outlet (21), the inner bottom end surface of the device shell (1) is provided with a droplet collection module (27) and a clamping jaw (28), and the inner bottom end surface of the device shell (1) is fixedly provided with a hole plate placement position (29).

2. The microdroplet culture device of claim 1, wherein, The oil pump injection pump (2), the liquid pump injection pump (3) and the gas pump injection pump (4) are uniformly distributed in the device shell (1).

3. The microdroplet culture device of claim 1, wherein, The first mounting frame (6) is provided with a culture bin (30), the culture bin (30) is provided with a heater (31), the top of the heater (31) is provided with a fan (5), the culture bin (30) is fixedly provided with a temperature sensor (26), and the culture bin (30) is provided with a first droplet disc (22), a second droplet disc (23), a third droplet disc (24) and a fourth droplet disc (25).

4. The microdroplet culture device of claim 1, wherein, The first pinch valve (8), the second pinch valve (9), the third pinch valve (10), the fourth pinch valve (11), the fifth pinch valve (12), the sixth pinch valve (13), the seventh pinch valve (14), the eighth pinch valve (15), the ninth pinch valve (16) and the tenth pinch valve (17) are the same in size.

5. The microdroplet culture device of claim 1, wherein The first pinch valve (8), the second pinch valve (9), the third pinch valve (10), the fourth pinch valve (11), the fifth pinch valve (12), the sixth pinch valve (13), the seventh pinch valve (14), the eighth pinch valve (15), the ninth pinch valve (16) and the tenth pinch valve (17) are uniformly distributed on the second mounting frame (7).

6. The microdroplet culture device of claim 3, wherein, The heater (31), the fan (5) and the temperature sensor (26) are arranged at the central part inside the culture bin (30).

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