Liquid drop generating device for low-temperature sample injection

Abstract

The utility model relates to the field of liquid drop microfluidics, in particular to a low-temperature sample injection liquid drop generating device. The utility model discloses a low-temperature sample injection liquid drop generation device which comprises a gas source, a pressure controller, a water-phase sample injection bottle, an oil-phase sample injection bottle, a water-phase sample injection pipe, an oil-phase sample injection pipe, a collecting bottle, a temperature control device and a liquid drop generation chip, and the liquid drop generation chip comprises at least one water-phase chamber, one oil-phase chamber and one sample discharge chamber. Temperature control devices are arranged at the sample introduction bottle and the collection bottle; the device is simple in structure, temperature adjustment can be carried out in the process from liquid drop generation to liquid drop collection completion, and the problem that reaction of liquid drops generated firstly and liquid drops generated later is not synchronous is solved, so that experimental errors are reduced, and experimental accuracy is improved.

Description

Technical Field

[0001] The utility model relates to the field of droplet microfluidics, and particularly to a droplet generating device with low-temperature sampling. Background Art

[0002] Based on the droplet microfluidics technology, an ultra-high-throughput single-cell sorting platform can generate a large number of micro-droplets per second. Cells are encapsulated in the micro-droplets, and biological and biochemical processes such as growth, lysis, metabolism, and reaction can be carried out. The cells are fully combined with the fluorescence sieves in the droplets to generate fluorescence signals of different intensities. Then, the micro-droplet sorting technology is used to sort the cells with low and high yields through the fluorescence signals, realizing the high-throughput of the sorting process.

[0003] Generally, after generating the droplets, further incubation is often required to enable a series of reactions inside the droplets for further screening. However, in the existing related technologies, due to the different droplet generation times, the droplets generated earlier have a longer growth or reaction time than the droplets generated later, and the variables cannot be controlled, thus affecting the accuracy of the experiment. Content of the Utility Model

[0004] To solve the above problems, the utility model provides a droplet generating device with low-temperature sampling, which includes a gas source, a pressure controller, an aqueous-phase sampling bottle, an oil-phase sampling bottle, an aqueous-phase sampling tube, an oil-phase sampling tube, a collection bottle, a temperature control device, a droplet generation chip, and a collection tube. The pressure controller is connected to the gas source to control the air pressure. The pressure controller and the aqueous-phase sampling bottle are connected through the aqueous-phase sampling tube. The oil-phase sampling bottle and the pressure controller are connected through the oil-phase sampling tube. The aqueous-phase sampling tube and the oil-phase sampling tube are respectively connected to the droplet generation chip, and are respectively used to introduce the aqueous-phase sample and the oil-phase sample into the droplet generation chip. The collection bottle and the droplet generation chip are connected through the collection tube. The droplets generated in the droplet generation chip flow into the collection bottle through the collection tube. The aqueous-phase sampling bottle, the oil-phase sampling bottle, and the collection bottle are placed in the temperature control device.

[0005] Further, the droplet generation chip includes at least one aqueous-phase chamber, at least one oil-phase chamber, and at least one sample discharge chamber. The aqueous-phase chamber is connected to the aqueous-phase sampling tube. The oil-phase chamber is connected to the oil-phase sampling tube. The sample discharge chamber is connected to the collection tube.

[0006] Further, an oil-phase pipeline, an aqueous-phase pipeline, and a sample discharge pipeline are arranged in the droplet generation chip. The oil-phase pipeline is connected to the oil-phase chamber. The aqueous-phase pipeline is connected to the aqueous-phase chamber. The end of the oil-phase pipeline is communicated with the end of the aqueous-phase pipeline, and then is connected to the sample discharge pipeline. The end of the sample discharge pipeline is connected to the sample discharge chamber.

[0007] Further, a serpentine structure is provided at the end of the oil phase pipeline, near the connection with the sample discharging pipeline.

[0008] Further, a serpentine structure is provided at the end of the water phase pipeline, near the connection with the sample discharging pipeline.

[0009] Further, a number of stoppers are provided in both the water phase chamber and the oil phase chamber.

[0010] Further, the temperature control device is a metal bath device.

[0011] The present utility model can control the temperature to be constant during the droplet generation process. When the temperature is set at 4°C or lower, it is not conducive to the growth or reaction of cells in the droplets. After all the droplets are generated and collected, appropriate environmental conditions are set to enable the cells in the droplets to start growing or reacting for subsequent experiments. In this way, the growth or reaction duration of the cells in the droplets can be controlled to be the same, increasing the reliability of the experiment. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The above and / or additional aspects and advantages of the present utility model will become apparent and be readily understood from the following description of the embodiments in conjunction with the accompanying drawings, in which:

[0013] Figure 1 is a schematic structural diagram of an embodiment of the present utility model;

[0014] Figure 2 is a schematic structural diagram of a droplet generation chip according to an embodiment of the present utility model;

[0015] Figure 3 is a partial enlarged view of an embodiment of the present utility model. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0016] The following describes in detail the embodiments of the present utility model. The examples of the embodiments are shown in the accompanying drawings, where the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary only for explaining the present utility model and should not be construed as limiting the present utility model.

[0017] In the description of the present utility model, it should be understood that the orientation or positional relationship indicated by the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", etc. is based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the present utility model and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, it should not be construed as a limitation to the present utility model. In addition, the terms "first" and "second" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly specifying the quantity of the indicated technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include one or more of the said features. In the description of the present utility model, "a plurality of" means two or more, unless otherwise specifically defined.

[0018] In the description of the present utility model, it should be noted that unless otherwise clearly specified and defined, the terms "installed", "connected" and "coupled" shall be understood in a broad sense. For example, it may be a fixed connection, a detachable connection or an integral connection; it may be a mechanical connection, an electrical connection or a connection capable of mutual communication; it may be directly connected or indirectly connected through an intermediate medium, and it may be the communication inside two elements or the interaction relationship between two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present utility model can be understood according to specific circumstances.

[0019] In the description of the present utility model, unless otherwise clearly specified and defined, the first feature being "on" or "under" the second feature may include the direct contact between the first and second features, or may include the situation where the first and second features are not in direct contact but in contact through additional features therebetween. Moreover, the first feature being "above", "over" and "on top of" the second feature includes that the first feature is directly above and obliquely above the second feature, or merely indicates that the horizontal height of the first feature is higher than that of the second feature. The first feature being "under", "beneath" and "underneath" the second feature includes that the first feature is directly below and obliquely below the second feature, or merely indicates that the horizontal height of the first feature is lower than that of the second feature.

[0020] The following disclosure provides many different embodiments or examples for implementing different structures of the present utility model. To simplify the disclosure of the present utility model, components and settings of specific examples are described below. Of course, they are only examples and are not intended to limit the present utility model. In addition, the present utility model may repeat reference numerals and / or reference letters in different examples. This repetition is for the purpose of simplification and clarity and does not itself indicate the relationship between various embodiments and / or settings discussed. In addition, the present utility model provides examples of various specific processes and materials, but those of ordinary skill in the art can be aware of the application of other processes and / or the use of other materials.

[0021] Referring to Figures 1-3 , a droplet generation device with low-temperature sampling according to an embodiment of the present utility model includes a gas source 1, a pressure controller 2, an aqueous-phase sampling bottle 3, an oil-phase sampling bottle 4, an aqueous-phase sampling tube 5, an oil-phase sampling tube 6, a collection bottle 7, a temperature control device 8, a droplet generation chip 9, and a collection tube 10.

[0022] The pressure controller 2 is connected to the gas source 1 to control the air pressure. In this embodiment, a common pump such as an injection pump, a peristaltic pump, a plunger pump, or a constant-pressure pump is selected as the pressure controller 2.

[0023] The pressure controller 2 and the aqueous-phase sampling bottle 3 are connected through the aqueous-phase sampling tube 5, and the oil-phase sampling bottle 4 and the pressure controller 2 are connected through the oil-phase sampling tube 6. The aqueous-phase sampling tube 5 and the oil-phase sampling tube 6 are respectively connected to the droplet generation chip 9 and are respectively used to introduce the aqueous-phase sample and the oil-phase sample into the droplet generation chip 9.

[0024] Specifically, the pressure controller 2 adjusts the flow rate of the gas to respectively push the liquids in the oil-phase sampling bottle 4 and the aqueous-phase sampling bottle 4 into the downstream oil-phase sampling tube 6 and aqueous-phase sampling tube 5, and thus into the droplet generation chip 9 for droplet generation operation.

[0025] The collection bottle 7 and the droplet generation chip 9 are connected through the collection tube 10. The droplets generated in the droplet generation chip 9 flow into the collection bottle 7 through the collection tube 10, thus completing the collection of the generated droplets.

[0026] The aqueous-phase sampling bottle 3, the oil-phase sampling bottle 4, and the collection bottle 7 are all placed in the temperature control device 8 to control the environmental temperature at 4°C or lower so that cells do not grow or react during the droplet generation process.

[0027] In the low-temperature sampling droplet generation device in the embodiments of the present utility model, by controlling the gas flow rate, the aqueous phase and the oil phase are pushed into the droplet generation chip 9 to generate "water-in-oil" droplets. The oil phase here is usually fluorinated oil, mineral oil, etc., and the aqueous phase is the cell solution, and the aqueous phase and the oil phase are immiscible. When it is not necessary for cells to grow or react during the droplet generation process, the temperature control device 8 can be used to control the ambient temperature at 4°C or below, so as to avoid affecting the experimental results due to the different generation times of different droplets during the droplet generation process.

[0028] Furthermore, the droplet generation chip 9 includes at least one aqueous phase chamber 91, at least one oil phase chamber 92, and at least one sample discharge chamber 93. The aqueous phase chamber 91 is connected to the aqueous phase sampling tube 5, the oil phase chamber 92 is connected to the oil phase sampling tube 6, and the sample discharge chamber 93 is connected to the collection tube 10.

[0029] As Figure 2 shown, an oil phase pipeline 95, a sample discharge chamber 94, and a sample discharge pipeline 96 are arranged in the droplet generation chip 9. The oil phase pipeline 95 is connected to the oil phase chamber 92, the sample discharge chamber 94 is connected to the aqueous phase chamber 91, the end of the oil phase pipeline 95 is communicated with the end of the sample discharge chamber 94, and then connected to the sample discharge pipeline 96. The end of the sample discharge pipeline 96 is connected to the sample discharge chamber 93.

[0030] As shown in the figure, the droplet generation chip 9 adopted in this embodiment is a "single aqueous phase" structure, that is, one kind of aqueous phase and one kind of oil phase act to generate droplets. In some cases, chips with a "multi-aqueous phase" or other structures can also be designed according to experimental requirements.

[0031] Specifically, the oil phase chamber 92 is connected to the oil phase pipeline 95, the aqueous phase chamber 91 is connected to the sample discharge chamber 94, the ends of the sample discharge chamber 94 and the oil phase pipeline 95 are communicated. After the oil phase and the aqueous phase form water-in-oil droplets at the connection point, they enter the sample discharge pipeline 96, flow into the sample discharge chamber 93 through the sample discharge pipeline 96, and finally enter the collection bottle 7.

[0032] Furthermore, a serpentine structure 98 is arranged at the end of the oil phase pipeline 95 close to the connection with the sample discharge pipeline 96.

[0033] Furthermore, a serpentine structure 98 is arranged at the end of the sample discharge chamber 94 close to the connection with the sample discharge pipeline 96.

[0034] As Figure 3 shown, the pipeline of the serpentine structure 98 is a folded-back pipeline, which can be U-shaped, S-shaped, etc. This design can make the fluid flow rate inside the pipeline stable and uniform, which is beneficial to the formation of droplets with consistent sizes.

[0035] Furthermore, a number of stoppers 97 are arranged in both the aqueous phase chamber 91 and the oil phase chamber 92.

[0036] The design of the stopper 97 first plays a filtering role, and secondly can support the chip chamber to prevent the chip substrate from deforming or collapsing.

[0037] Furthermore, the temperature control device 8 is a metal bath device.

[0038] In some embodiments, the temperature control device 8 can adopt a metal bath device, in which the aqueous sample injection bottle 3, the oil-phase sample injection bottle 4 and the collection bottle 7 are placed. The metal bath device has the advantages of accurate and uniform temperature control and can better achieve the temperature control effect.

[0039] Although the embodiments of the present invention have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principle and spirit of the present invention. The scope of the present invention is defined by the claims and their equivalents.

Claims

1. A droplet generating device for low temperature injection, characterized in that: It includes an air source, a pressure controller, an aqueous phase injection bottle, an oil phase injection bottle, an aqueous phase injection tube, an oil phase injection tube, a collecting bottle, a temperature control device, a droplet generation chip, and a collecting tube. The pressure controller is connected to the air source to control the air pressure. The pressure controller is connected to the aqueous phase injection bottle through the aqueous phase injection tube. The oil phase injection bottle is connected to the pressure controller through the oil phase injection tube. The aqueous phase injection tube and the oil phase injection tube are respectively connected to the droplet generation chip and are used to respectively inject aqueous phase samples and oil phase samples into the droplet generation chip. The collecting bottle is connected to the droplet generation chip through the collecting tube. Droplets generated in the droplet generation chip flow into the collecting bottle through the collecting tube. The aqueous phase injection bottle, the oil phase injection bottle, and the collecting bottle are placed in the temperature control device.

2. A droplet generating device for low temperature injection according to claim 1, characterized in that: The droplet generation chip comprises at least one water phase chamber, at least one oil phase chamber and at least one sample row chamber, wherein the water phase chamber is connected to the water phase injection tube, the oil phase chamber is connected to the oil phase injection tube, and the sample row chamber is connected to the collection tube.

3. A cryogenic droplet generating device according to claim 2, characterized in that: The droplet generation chip is provided with an oil phase pipeline, a water phase pipeline and a sample discharge pipeline, the oil phase pipeline is connected to the oil phase chamber, the water phase pipeline is connected to the water phase chamber, the oil phase pipeline is communicated with the end of the water phase pipeline, and then connected to the sample discharge pipeline, and the end of the sample discharge pipeline is connected to the sample discharge chamber.

4. A cryogenic droplet generating device according to claim 3, characterized in that: A serpentine structure is arranged at the end of the oil phase pipeline, near the connection with the sample discharge pipeline.

5. A cryogenic droplet generating device according to claim 3, characterized in that: A serpentine structure is arranged at the end of the water phase pipeline, near the connection with the sample discharge pipeline.

6. A cryogenic droplet generating device according to claim 2, characterized in that: A plurality of blocks are arranged in the water phase chamber and the oil phase chamber.

7. A droplet generating device for low temperature injection according to claim 1, characterized in that: The temperature control device is a metal bath device.

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