A multifunctional microchannel array droplet generator and methods of use thereof

Abstract

The application discloses a multifunctional microchannel array droplet generator, which comprises a sample barrel, a water phase channel is arranged in the middle of the sample barrel and penetrates the sample barrel from top to bottom, the water phase channel is used for containing a water phase or as a water phase inlet; a chip is bonded and connected between the bottom surface of the sample barrel and the upper end surface of the chip, a plurality of microchannel arrays are engraved around the outer periphery of the chip, a solution main flow channel is arranged in the middle of the chip and is in communication with the water phase channel, and a plurality of uniformly arranged solution auxiliary flow channels are arranged outward and diverge from the outer periphery of the circular solution main flow channel. The application can realize the method for high-throughput and accurate preparation of uniform droplets through handheld injection, injection pump injection or batch centrifugation, overcome the problem that the cost of the existing droplet generation device and control instrument is high, and can generate droplets at any time and anywhere, high-throughput droplet generation, a large number of parallel experiments can be carried out, and the demand of different application scenes can be met.

Description

TECHNICAL FIELD

[0001] The present application relates to a droplet generation technology, in particular to a multifunctional microchannel array droplet generator and a method thereof. BACKGROUND

[0002] Stable and uniform water-in-oil droplet micro-reactors have broad application prospects in the biomedical field such as single-cell sequencing, molecular diagnosis, and hydrogel microsphere synthesis. Microfluidic technology is a general method for forming uniform droplets by controlling fluid movement in microchannels. However, the existing microfluidic chips usually require additional expensive and bulky auxiliary equipment, and the operation method is relatively complex, thereby increasing the cost and labor cost, and limiting the application scenarios. Therefore, it is necessary to develop a simple and convenient multifunctional droplet generation method to reduce the cost and meet the use requirements of different application scenarios.

[0003] Therefore, the auxiliary equipment matched with the droplet generator in the prior art is expensive and has limited use environment, which cannot meet the technical requirements of use in different scenarios. SUMMARY

[0004] The present application aims to provide a multifunctional microchannel array droplet generator and a method thereof to solve the problems raised in the background.

[0005] To achieve the above-mentioned purpose, the present application provides the following technical scheme: a multifunctional microchannel array droplet generator, characterized in that it comprises:

[0006] A sample barrel body, a water phase channel is provided in the middle part of the sample barrel body, the water phase channel is used for containing water phase or as a water phase inlet;

[0007] A chip is bonded and connected between the upper end face of the chip and the bottom surface of the sample barrel body, a plurality of microchannel arrays are engraved around the outer periphery of the chip, a solution main flow channel is provided in the middle part of the chip and is in communication with the water phase channel, and a plurality of uniformly arranged solution auxiliary flow channels are provided on the outer periphery of the solution main flow channel.

[0008] Preferably, the microchannel array comprises protrusions arranged in sequence and spaced apart around the outer periphery of the chip, a microchannel is formed between any two adjacent protrusions, the end of the microchannel is trumpet-shaped, and the trumpet mouth of the microchannel is arranged outward.

[0009] Preferably, the upper end of the protrusion is higher than the upper end face of the chip, a clamping block is arranged at each corner of the six edges of the chip, a clamping groove is arranged on the bottom surface of the sample barrel body corresponding to the clamping block, and the clamping block is clamped into the clamping groove to realize connection.

[0010] The micro-channel and the bottom surface of the sample barrel form a gap for the water phase in the sample solution to disperse into the oil phase.

[0011] Preferably, the chip is made of silicon and has a hexagonal prism structure with six edges, each solution auxiliary flow channel is arranged at the middle of each edge and forms a merging groove for the sample solution to merge and flow between the rear end surface of the convex block and the solution auxiliary flow channel.

[0012] The sample barrel is made of plastic and has a hexagonal prism structure, and the shape of the sample barrel is matched with the chip.

[0013] Preferably, the water phase channel is provided with a water phase cup for containing the sample solution.

[0014] Preferably, the chip adopts the gradient emulsification principle, and the size of the generated droplets is matched with the shape and size of the micro-channel.

[0015] The application also discloses a use method of the multifunctional micro-channel array droplet generator.

[0016] The chip part of the droplet generator is immersed in a centrifuge tube containing the oil phase.

[0017] The sample solution is added to the water phase channel as the water phase.

[0018] The water phase in the water phase channel is extruded into the chip by the pressure generating device, the water phase diffuses outward along the solution main flow channel of the chip to each solution auxiliary flow channel, and then diffuses to the micro-channel array through the solution auxiliary flow channel, and then disperses into the oil phase in the centrifuge tube through the micro-channel array to generate droplets.

[0019] Preferably, the pressure generating device is a micro-syringe, the outlet end of the micro-syringe is connected to the water phase channel by holding the micro-syringe by hand, the water phase is extruded into the chip by the pressure generated by injection, and the droplets are generated by dispersing into the oil phase.

[0020] Preferably, the pressure generating device is an injection pump and a micro-syringe, the injection pump is connected to the micro-syringe, the outlet end of the micro-syringe is connected to the water phase channel, the water phase is extruded into the chip by the pressure generated by the injection pump to the micro-syringe, and the droplets are generated by dispersing into the oil phase.

[0021] Preferably, the pressure generating device is a centrifuge, a plurality of mounting slots are uniformly arranged on the centrifuge, the chip is immersed in the centrifugal tube containing oil to form a droplet generating device, and the water phase is added through the water phase channel, the droplet generating device is put into the mounting slots in batches, the centrifuge is started, and the water phase is thrown into the oil phase by the centrifugal force of the centrifuge to generate droplets in batches.

[0022] Compared with the prior art, the droplet generator of the application is simple and efficient, can realize the method of high-throughput and accurate preparation of uniform droplets through handheld injection, injection pump injection or batch centrifugation, overcome the problem of high cost of the existing droplet generating device and its control instrument, and can generate droplets anytime and anywhere, high-throughput droplet generation, a large number of parallel experiments can also be carried out, meet the needs of different application scenarios, and can ensure the uniformity of the size of the generated droplet particles, effectively improve the CV value of the size of the droplet particles. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 The structure diagram of the chip in the droplet generator of the application is shown in the figure.

[0024] Figure 2 The structure diagram of the sample barrel body in the droplet generator of the application is shown in the figure.

[0025] Figure 3 The structure diagram of the droplet generator of the application is shown in the figure.

[0026] Figure 4 The installation structure diagram of the centrifugal tube matched with the droplet generator in the generation of droplets of the application is shown in the figure.

[0027] Figure 5 The installation structure diagram of the water phase cup and the centrifugal tube matched with the droplet generator in the generation of droplets of the application is shown in the figure.

[0028] Figure 6 The structure diagram of the handheld microsyringe in the first embodiment of the application is shown in the figure.

[0029] Figure 7 The structure diagram of the injection pump in the second embodiment of the application is shown in the figure.

[0030] Figure 8 The structure diagram of the centrifugal pump in the third embodiment of the application is shown in the figure.

[0031] Figure 9 The droplet morphology produced by different experimenters through handheld injection in the first embodiment is shown in the figure.

[0032] Figure 10 The droplet morphology produced at different flow rates in the second embodiment is shown in the figure.

[0033] Figure 11 Droplet morphology produced for a second embodiment with a different oil phase;

[0034] Figure 12 Droplet morphology produced for a third embodiment with a different centrifugation rate. DETAILED DESCRIPTION

[0035] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by a person of ordinary skill in the art without creative effort fall within the protection scope of the present application.

[0036] Referring to Figures 1-3 The present application provides a multifunctional microchannel array droplet generator, comprising a sample barrel 2 and a chip 1, wherein the sample barrel 2 is preferably made of plastic material and has a hexagonal prism structure, and a water phase channel 4 is formed in the middle of the sample barrel 2 and penetrates the sample barrel 2 from top to bottom. The water phase channel 4 can be used to hold water phase or as a water phase inlet after being bonded with the chip 1, and the droplet generating device has higher integration.

[0037] In another embodiment, referring to Figure 5 A water phase cup 8 for holding sample solution can also be installed in the water phase channel 4, that is, the water phase cup is inserted into the water phase channel 4, and the upper end of the water phase cup 8 protrudes outward with an outer rim 801, and the diameter of the outer rim 801 is greater than the diameter of the water phase channel 4. Therefore, when the water phase cup 8 is inserted into the water phase channel 4, the outer rim 801 plays a role in limiting the position of the water phase cup 8, and at the same time can prevent the water phase cup 8 from falling into the water phase channel 4. The sample solution can also be referred to as water phase, and the water phase cup 8 can be assembled with a standard syringe port. The characteristics and innovations are portability, high integration, and matching with commonly used laboratory syringes and centrifuge tubes.

[0038] The upper end surface of the chip 1 is bonded and connected with the bottom surface of the sample barrel 2, and a plurality of microchannel arrays are engraved around the outer periphery of the chip 1. A solution main flow channel 31 is formed in the middle of the chip 1 and is in communication with the water phase channel 4. The sample solution flows into the solution main flow channel 3 through the water phase channel 4. A plurality of uniformly arranged solution auxiliary flow channels 31 are formed on the outer periphery of the circular solution main flow channel 3 and diverge outward. The sample solution is dispersed outward through the main flow channel 3 and flows into each solution auxiliary flow channel 31.

[0039] The micro-channel array comprises convex blocks 5 arranged in sequence and spaced apart around the outer periphery of the chip 1, and a micro-channel 6 is formed between any two adjacent convex blocks 5, the micro-channel 6 is used for flowing sample solution and forming generated droplets, the end of the micro-channel 6 is trumpet-shaped, and the trumpet mouth of the micro-channel 6 is outwardly arranged. The shape and size of the micro-channel 6 determine the size of the generated droplet particles, which can effectively ensure the uniformity of the droplet size, and effectively improve the CV value of the droplet particle size. The chip 1 adopts the gradient emulsification principle, the size of the generated droplet is matched with the shape and size of the micro-channel 6, and theoretically the droplet size is only related to the shape of the water phase channel micro-channel 6. Therefore, as long as the hydrophobic treatment inside the chip 1 is good, the size of the droplet will not be affected by the water phase flow rate or pressure.

[0040] The upper end of the convex block 5 is higher than the upper end surface of the chip 1, and the chip 1 is provided with a clamping block 71 at each corner of the four corners, and the bottom surface of the sample barrel body 2 is provided with a clamping groove corresponding to the clamping block 71, and the clamping block 71 is clamped into the clamping groove to realize connection; since the upper end surface of the convex block 5 is higher than the upper end surface of the chip 1 by a distance, after the chip 1 and the sample barrel body 2 are connected, the micro-channel 6 and the bottom surface of the sample barrel body 2 form a gap for the water phase in the sample solution to disperse into the oil phase, that is, a channel for forming droplets is formed.

[0041] In the present application, the chip 1 is made of silicon, the chip 1 is a hexagonal prism structure surrounded by six edges 7, the shape of the sample barrel body 2 is matched with the chip 1, each solution auxiliary flow channel 31 is arranged at the middle of each edge 7, and a confluence groove 32 for the confluence of sample solution is formed between the rear end surface of the convex block 5, the confluence groove 32 communicates with the solution auxiliary flow channel 31, so that the sample solution flows into the confluence groove 32 through each solution auxiliary flow channel 31, and then flows into the micro-channel 6 through the confluence groove 32, forming smooth flow of the water phase.

[0042] The design of the droplet generator of the device is based on the step emulsification principle, which is a preparation method for spontaneously forming droplets induced by Laplace pressure difference of geometric shape, mainly driven by interfacial tension, rather than high-energy shear stress system. Theoretically, the size of the droplet is independent of the flow rate of the continuous phase and the dispersed phase, so it has the characteristics of low shear, insensitivity to flow rate and simple structure easy to integrate. Therefore, the droplet generator of the device can produce droplets with uniform particles under different pressures and different rates.

[0043] Referring to the drawings Figure 4 - the drawings Figure 8 The present application also discloses a use method of the multifunctional micro-channel array droplet generator, which adopts the multifunctional micro-channel array droplet generator described above, and comprises the following steps:

[0044] The chip 1 of the droplet generator is partially immersed in a centrifuge tube 9 containing the oil phase, preferably a 1.5ml centrifuge tube;

[0045] The sample solution is added to the water phase channel 4 as the water phase;

[0046] The water phase in the water phase channel 4 is extruded into the chip 1 by the pressure generating device, and the water phase diffuses outward along the main flow channel 3 of the chip to each auxiliary flow channel 31, and then diffuses into each microchannel 6 of the microchannel array through the auxiliary flow channel 31, and then disperses into the oil phase in the centrifuge tube 9 through the microchannel array, and under the pressure of the pressure generating device, droplets are generated.

[0047] In the present application, the pressure generating device can be flexibly selected as needed, suitable for various environments, and also suitable for various extrusion pressures, and can generate droplets of basically uniform size.

[0048] Referring to Figure 6 , the pressure generating device is a microsyringe 81, and in use, the chip 1 and the like are immersed in the centrifuge tube 9 containing the oil, and the outlet end of the microsyringe 81 is connected to the water phase channel 4, and the water phase is extruded into the chip 1 by the pressure generated by the injection, and then sequentially passes through the main flow channel 3---auxiliary flow channel 31---converging groove 32---microchannel 6, and then disperses into the oil phase through the microchannel 6 to generate droplets.

[0049] The biggest innovation of using the handheld microsyringe 81 is that it can stably generate droplets in a handheld manner, without the need for a bulky auxiliary device (such as a syringe pump), and without the need to generate droplets in a fixed scene, while reducing the use cost.

[0050] Referring to Figure 9 It can be seen that different experimenters produce droplets by hand injection, and the droplet size is uniform, the error of different personnel is small, the operation is simple, anyone with any education can apply and operate, and it is self-explanatory without professional training, which greatly expands the application scene.

[0051] Referring to Figure 7 , the pressure generating device is a syringe pump 82 used in combination with the microsyringe 81, and the syringe pump 82 is connected to the microsyringe 81, so that the pressure source is not provided by hand, and the pressure is automatically applied to the microsyringe 81 by the syringe pump 82.

[0052] The outlet end of the microsyringe 81 is connected to the water phase channel 4, and the water phase is extruded into the chip 1 by the pressure generated by the syringe pump 82 on the microsyringe 81, and then sequentially passes through the main flow channel 3---auxiliary flow channel 31---converging groove 32---microchannel 6, and then disperses into the oil phase 91 in the centrifuge tube 9 through the microchannel 6 to generate droplets.

[0053] The biggest innovation of this embodiment is that it can work automatically without manual power supply, improving the intelligence and enabling high-throughput continuous droplet generation, with a maximum droplet generation rate of tens of mL / h.

[0054] Referring to Figure 10 It can be seen that the injection pump 82 can be used to continuously produce droplets at a high throughput. When the flow rate is low (less than 1000 μL / h), the size of the generated droplets is uniform. When the flow rate continues to increase, the size of the droplets increases slightly, and the uniformity of the size decreases slightly. It should be pointed out that the throughput of the formed droplets can be further improved by increasing the number of internal water phase channels of the chip 1 to meet the demand of large-scale production of droplets in industry.

[0055] In addition, the dispersed phase has good compatibility with mineral oil or fluorinated oil, as shown in Figure 11 When 7500 fluorinated oil is used as the dispersed phase, the uniformity of the droplets can also be ensured.

[0056] Referring to Figure 8 The pressure generating device is a centrifuge 83, and a plurality of installation slots 84 are uniformly arranged on the centrifuge 83. The installation slots 84 are used for inserting the packaged droplet generating device, and the chip 1 is immersed in the oil-containing centrifuge tube 9 to form a droplet generating device. The water phase is added through the water phase channel 4, and the droplet generating device is placed in the installation slot 84 in batches. The centrifuge 83 is started, and the water phase is thrown into the oil phase 91 by the centrifugal force of the centrifuge 83 to generate droplets in batches. The specific liquid flow is still through the main flow channel 3---the auxiliary flow channel 31---the merging groove 32---the microchannel 6 in sequence, and the microchannel 6 is dispersed into the oil phase 91 in the centrifuge tube 9 to generate droplets.

[0057] The biggest innovation of this embodiment is that it can generate droplets by means of the existing centrifuge in the laboratory, without the need to purchase additional matching devices, thereby saving the use cost. In addition, the experiment can be carried out in batches by the centrifuge, thereby saving the time cost and labor cost and improving the production efficiency.

[0058] The method of generating droplets by centrifugation can simultaneously produce a large number of droplets, as shown in Figure 12 When the centrifugal rate is 100-200 g, only 1 minute of centrifugation is needed, and 30 μL of water phase can completely generate droplets, and the uniformity and stability of the droplets can be ensured. This method can meet various parallel experiments, and greatly reduces the operation cost and time cost of the experiment.

[0059] In the above three embodiments, the embodiment in which the water phase cup 8 is installed in the water phase channel 4 is also applicable.

[0060] In summary, the application discloses a simple and efficient method for preparing uniform droplets with high throughput and precision, which can be realized by handheld injection, injection pump injection or batch centrifugation, and overcomes the problem of high cost of existing droplet generation devices and their matching instruments, and can generate droplets anytime and anywhere, high-throughput and fast generation of droplets, and can also perform a large number of parallel experiments, meet the needs of different application scenarios, and has practical application value in molecular diagnosis.

[0061] In the description of the present application, it should be noted that the terms "upper", "lower", "front", "back", "inner", "outer" and the like indicate the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, therefore cannot be understood as a limitation on the present application.

[0062] In the description of the present application, it should be noted that unless otherwise explicitly specified and limited, the terms "mounting", "connecting", "connecting" should be understood broadly, for example, it can be fixedly connected, or it can be detachably connected, or integrally connected, it can be mechanically connected, or it can be electrically connected, it can be directly connected, or it can be indirectly connected through an intermediate medium, it can be the communication inside two elements. For those skilled in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances.

[0063] The preferred embodiments of the present application are described above, but the present application is not limited to the above embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the purpose of the present application.

Claims

1. A multifunctional microchannel array droplet generator, characterized in that, comprising: a sample barrel (2) having a water phase channel (4) passing through the middle part of the sample barrel (2) from top to bottom, which is used to hold the water phase or as the water phase inlet; a chip (1) bonded to the bottom surface of the sample barrel (2), a plurality of microchannel arrays are engraved around the outer periphery of the chip (1), and a solution main flow channel (3) is provided in the middle part of the chip (1) and is in communication with the water phase channel (4), and a plurality of solution auxiliary flow channels (31) are evenly arranged and diverge outward from the outer periphery of the solution main flow channel (3), the microchannel array comprises a plurality of protrusions (5) arranged in sequence around the outer periphery of the chip (1), and a microchannel (6) is formed between any two adjacent protrusions (5), the end of the microchannel (6) is trumpet-shaped, and the trumpet mouth of the microchannel (6) is outwardly arranged, the upper end of the protrusion (5) is higher than the upper end surface of the chip (1), a clamping block (71) is arranged at each corner of the six edges of the chip (1), a clamping groove is arranged on the bottom surface of the sample barrel (2) corresponding to the clamping block (71), and the clamping block (71) is clamped into the clamping groove to achieve connection; the microchannel (6) and the bottom surface of the sample barrel (2) form a gap for the water phase in the sample solution to disperse into the oil phase. 2.The multifunctional microchannel array droplet generator according to claim 1, characterized in that, the chip (1) is made of silicon, the chip (1) is a hexagonal prism structure enclosed by six edges (7), each solution auxiliary flow channel (31) is arranged in the middle of each edge (7) and forms a merging groove (32) between the rear end surface of the protrusion (5) for the sample solution to flow, and the merging groove (32) is in communication with the solution auxiliary flow channel (31); the sample barrel (2) is made of plastic and has a hexagonal prism structure, and the shape of the sample barrel (2) is adapted to the shape of the chip (1). 3.The multifunctional microchannel array droplet generator according to claim 1, characterized in that, a water phase cup for holding a sample solution is installed in the water phase channel (4). 4.The multifunctional microchannel array droplet generator according to claim 2, characterized in that, the chip (1) adopts the principle of gradient emulsification, and the size of the generated droplets is adapted to the shape and size of the microchannel (6). 5.A method for using a multifunctional microchannel array droplet generator, characterized in that, the multifunctional microchannel array droplet generator according to any one of claims 1-4 is used, comprising the following steps: immersing the chip (1) part of the droplet generator into a centrifuge tube (9) containing an oil phase; adding a sample solution as a water phase to the water phase channel (4); The water phase in the water phase channel (4) is extruded into the chip (1) by the pressure generating device, and the water phase diffuses outward along the solution main flow channel (3) of the chip to each solution auxiliary flow channel (31), and then diffuses into the microchannel array through the solution auxiliary flow channel (31), and then disperses into the oil phase in the centrifuge tube (9) through the microchannel array, to generate droplets.

6. The method for using the multifunctional microchannel array droplet generator according to claim 5, wherein the pressure generating device is a microsyringe (81), the outlet end of the microsyringe is connected to the water phase channel (4) by hand-holding the microsyringe, and the water phase is extruded into the chip (1) by the pressure generated by injection and dispersed into the oil phase to generate droplets.

7. The method for using the multifunctional microchannel array droplet generator according to claim 5, wherein the pressure generating device is an injection pump (82) and a microsyringe (81), the injection pump (82) is connected to the microsyringe (81), the outlet end of the microsyringe (81) is connected to the water phase channel (4), and the water phase is extruded into the chip (1) by the pressure generated by the injection pump (82) on the microsyringe (81) and dispersed into the oil phase to generate droplets.

8. The method for using the multifunctional microchannel array droplet generator according to claim 5, wherein the pressure generating device is a centrifuge (83), a plurality of mounting slots (84) are uniformly arranged on the centrifuge (83), the chip (1) is immersed in the centrifuge tube (9) containing oil to form a droplet generating device, the water phase is added through the water phase channel (4), the droplet generating device is placed in the mounting slots (84) in batches, the centrifuge (83) is started, and the water phase is spun into the oil phase by the centrifugal force of the centrifuge (83) to generate droplets in batches. ​ ​ ​

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