CN110624616A - Three-dimensional microfluidic device and method for high-throughput micro-droplet generation - Google Patents
Three-dimensional microfluidic device and method for high-throughput micro-droplet generation Download PDFInfo
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Abstract
A three-dimensional microfluidic device and method for generating high-flux micro-droplets, the device comprises a closed flow channel structure formed by bonding a bottom layer, a middle layer and an upper layer PDMS flow channel, wherein the closed flow channel structure is provided with a dispersed phase inlet joint, a continuous phase inlet joint and a collection outlet joint; fixing a three-dimensional microfluidic device on an objective table of a microscope, starting an injection pump, firstly adjusting a dispersion phase and a continuous phase to corresponding flow velocities through the injection pump, and shearing the dispersion phase into micro-droplets by the continuous phase at positions where T-shaped micro-droplets in the device generate a flow channel array; the invention can overcome the defect of slow micro-droplet generation rate of the current two-dimensional flow channel, realizes the generation of high-flux micro-droplets, has the integral structure of PDMS, is very firm after being mutually bonded, avoids the occurrence of cracking between the flow channels due to overlarge input fluid pressure, and has transparent PDMS and easier observation.
Description
Technical Field
The invention relates to the technical field of microfluidics, in particular to a three-dimensional microfluidic device and a method for generating high-flux micro-droplets.
Background
Droplet microfluidic platforms have become biological and chemicalImportant tools in the fields of science, medicine and the like are used as important technologies in a droplet microfluidic platform, a micro-droplet generation method is important, and different methods have great difference in droplet generation rate. At present, the generation method of micro-droplets mainly includes a co-flow method, a flow-type convergence method, a T-type flow channel method and a derivation method based on the above methods, and by adjusting the input pressure of a gas phase (dispersed phase) and a liquid phase (continuous phase), the gas phase can be rapidly cut into micro-bubbles by the liquid phase, and the liquid phase is usually oil and aqueous solution. For example, by the parallel configuration of the focusing microfluidic device and the T-type microchannel device, 10 can be produced in less than one hour11Bubbles (Jeong H, Chen Z, Yadavali S, et al, Large-scale production of compounds using matched microfluidics for influencing external reactions [ J]Lab on a Chip,2019,19, 665-. For example, CN109701430A discloses a method for controlling a T-type micro-fluidic chip to generate micro-bubbles by using a vibrating pipeline, which can realize the formation of a monodisperse micro-bubble sequence. CN 105688721 a discloses a microfluidic chip for generating spherical microbubbles, which can generate microbubbles with different diameters by adjusting the liquid flow rate. Chinese patent (publication No. CN 109908983A) is a microfluidic chip with a three-dimensional conical structure and used for micro-droplet high-proportion splitting extraction, and relates to a microfluidic chip with a three-dimensional conical structure. CN107519958A discloses a two-dimensional focusing microfluidic device. However, in the above methods, the microfluidic devices used are mostly two-dimensional flow channels, the generation rate thereof is limited to a certain extent, and the method is not suitable for the generation of large-flux micro-droplets, and particularly when large-scale material manufacturing is required, the generation rate is difficult to meet the requirement.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a three-dimensional microfluidic device and a method for generating high-flux micro-droplets, the high-flux generation of the micro-droplets is realized through the three-dimensional array arrangement of a T-shaped flow channel structure, the size of the micro-droplets can be controlled and adjusted through the input pressure of a disperse phase inlet and a continuous phase inlet, the device is made of a full PDMS material, the whole body is transparent, the three-dimensional observation is convenient, the bonding among the structures of the device is firm, the cracking is not easy to occur, and the device is not only suitable for the high-flux generation of the micro-droplets, but also suitable for the high-.
In order to achieve the purpose, the invention adopts the following technical scheme:
a three-dimensional microfluidic device for generating high-flux micro-droplets comprises a closed flow channel structure formed by bonding a bottom PDMS flow channel 400, a middle PDMS flow channel 500 and an upper PDMS flow channel 600, wherein the closed flow channel structure is provided with a dispersed phase inlet connector 100, a continuous phase inlet connector 300 and a collection outlet connector 200;
the bottom PDMS flow channel 400 includes two sets of T-type droplet generation flow channel arrays, where the first set of T-type droplet generation flow channel array includes a first dispersed phase flow channel 402, a first continuous phase flow channel 401, a second continuous phase flow channel 405, and a second dispersed phase flow channel 406, an inlet end of the first dispersed phase flow channel 402 intersects with an inlet end of the second dispersed phase flow channel 406 and is connected to a first dispersed phase inlet 407, an outlet end of the first dispersed phase flow channel 402 communicates with the middle of the first continuous phase flow channel 401, an inlet end of the first continuous phase flow channel 401 intersects with an inlet end of the second continuous phase flow channel 405 and is connected to the first continuous phase inlet 414, and an outlet end of the first continuous phase flow channel 401 is connected to the first delivery outlet 403; the outlet end of the second dispersed phase flow channel 406 is communicated with the middle part of the second continuous phase flow channel 405, and the outlet end of the second continuous phase flow channel 405 is connected with the second conveying outlet 404;
the second group of T-shaped micro-droplet generation flow channel array comprises a third dispersed phase flow channel 408, a third continuous phase flow channel 410, a fourth dispersed phase flow channel 412 and a fourth continuous phase flow channel 413, wherein the inlet end of the third dispersed phase flow channel 408 is connected with the first dispersed phase inlet 407 after meeting with the inlet end of the fourth dispersed phase flow channel 412, the outlet end of the third dispersed phase flow channel 408 is communicated with the middle part of the third continuous phase flow channel 410, the inlet end of the third continuous phase flow channel 410 is connected with the first continuous phase inlet 414 after meeting with the inlet end of the fourth continuous phase flow channel 413, and the outlet end of the third continuous phase flow channel 410 is connected with a third conveying outlet 409; the outlet end of the fourth dispersed phase flow channel 412 communicates with the middle of the fourth continuous phase flow channel 413, and the outlet end of the fourth continuous phase flow channel 413 is connected to the fourth delivery outlet 411.
The middle PDMS runner 500 includes two sets of T-shaped micro-droplet generation runner arrays, where the first set of T-shaped micro-droplet generation runner array includes a fifth dispersed phase runner 503, a fifth continuous phase runner 502, a sixth continuous phase runner 501 and a sixth dispersed phase runner 507, an inlet end of the fifth dispersed phase runner 503 is connected to the second dispersed phase inlet 508 after meeting with an inlet end of the fifth dispersed phase runner 507, an outlet end of the fifth dispersed phase runner 503 is connected to the middle of the fifth continuous phase runner 502, an inlet end of the fifth continuous phase runner 502 is connected to the second continuous phase inlet 516 after meeting with an inlet end of the sixth continuous phase runner 501, and an outlet end of the fifth continuous phase runner 502 is connected to the fifth delivery outlet 504; an outlet end of the sixth dispersed phase flow channel 507 is communicated with the middle part of the sixth continuous phase flow channel 501, an outlet end of the sixth continuous phase flow channel 501 is connected with a sixth delivery outlet 507, and a fifth delivery outlet 504 and a sixth delivery outlet 506 are converged at a seventh delivery outlet 505 through the flow channels;
the second group of T-shaped micro droplet generation flow channel arrays includes a seventh dispersed phase flow channel 509, a seventh continuous phase flow channel 515, an eighth continuous phase flow channel 514 and an eighth dispersed phase flow channel 513, an inlet end of the seventh dispersed phase flow channel 509 is connected to the second dispersed phase inlet 508 after meeting with an inlet end of the eighth dispersed phase flow channel 513, an outlet end of the seventh dispersed phase flow channel 509 is communicated with the middle of the seventh continuous phase flow channel 515, an inlet end of the seventh continuous phase flow channel 515 is connected to the second continuous phase inlet 516 after meeting with an inlet end of the eighth continuous phase flow channel 514, and an outlet end of the seventh continuous phase flow channel 515 is connected to the eighth delivery outlet 510; an outlet end of the eighth dispersed phase flow channel 513 is communicated with a middle portion of the eighth continuous phase flow channel 514, an outlet end of the eighth continuous phase flow channel 514 is connected with the ninth delivery outlet 512, and the eighth delivery outlet 510 and the ninth delivery outlet 512 are merged at the tenth delivery outlet 511 through the flow channel.
The top PDMS flow channel 600 comprises a third dispersed phase inlet 604, a third continuous phase inlet 601 and a micro-droplet collecting port 605, wherein the third dispersed phase inlet 604 is connected to the dispersed phase inlet connector 100, and the third continuous phase inlet 601 is connected to the continuous phase inlet connector 300; the inlet end of the micro-droplet collecting port 605 is connected to the outlet ends of the eleventh and twelfth delivery outlets 603 and 606 through the first and second delivery flow channels 602 and 607.
The height of the flow channel of the closed flow channel structure is 60 micrometers, and the height of the flow channel is equal to the height of the flow channel except for a fifth conveying outlet 504, a sixth conveying outlet 506, a seventh conveying outlet 505, an eighth conveying outlet 510, a ninth conveying outlet 512, a second dispersed phase inlet 508 and a tenth conveying outlet 511, wherein a second continuous phase inlet 516, a third dispersed phase inlet 604, a micro-droplet collecting port 605 and a third continuous phase inlet 601 are all through holes, the aperture of the through holes is the same, and all other inlets and outlets are non-through holes.
The relative position relationship among the bottom layer PDMS flow channel 400, the middle layer PDMS flow channel 500, and the upper layer PDMS flow channel 600 is as follows: the lower surface of the middle PDMS runner 500 without a runner is bonded to the upper surface of the bottom PDMS runner 400 with a runner, and the lower surface of the top PDMS runner 600 is bonded to the upper surface of the middle PDMS runner 500 with a runner; the central axes of the first dispersed phase inlet 407, the second dispersed phase inlet 508, the third dispersed phase inlet 604 and the dispersed phase inlet joint 100 are coaxial and are communicated with each other; the central axes of the first continuous phase inlet 414, the second continuous phase inlet 516, the third continuous phase inlet 601 and the continuous phase inlet connector 300 are coaxial and are communicated with each other; the first delivery outlet 403 and the fifth delivery outlet 504, the second delivery outlet 404 and the sixth delivery outlet 506, the third delivery outlet 409 and the eighth delivery outlet 510, the fourth delivery outlet 411 and the ninth delivery outlet 512, the seventh delivery outlet 505 and the eleventh delivery outlet 603, and the tenth delivery outlet 511 and the twelfth delivery outlet 606 are coaxial with each other and are communicated with each other.
The micro-droplet generation method for the three-dimensional micro-fluidic device for high-throughput micro-droplet generation comprises the following steps:
1) fixing the three-dimensional microfluidic device for high-flux micro-droplet generation on an objective table of a microscope, and ensuring that two groups of T-shaped micro-droplet generation flow channel array parts of each middle-layer PDMS flow channel 500 are positioned in a field of view of the microscope and have no inclination through objective observation;
2) respectively connecting a disperse phase inlet connector 100, a continuous phase inlet connector 300 and a collection outlet connector 200 with a disperse phase solution storage bottle, a continuous phase solution storage bottle and a micro-droplet collection container on a nitrogen pressure injection pump through Teflon catheters;
3) and starting the injection pump, adjusting the dispersed phase and the continuous phase to corresponding flow rates through the injection pump, and shearing the dispersed phase into micro droplets by the continuous phase at the positions where the T-shaped micro droplets generate the flow channel array.
Compared with the prior art, the invention has the following beneficial effects:
(1) the invention can overcome the defect of slow micro-droplet generation rate of the current two-dimensional flow channel, and the device can accumulate and amplify the T-shaped micro-droplet generation flow channel array generated by micro-droplets, thereby realizing the generation of high-flux micro-droplets.
(2) The structure of each layer of flow channel is PDMS, the flow channels are firmly bonded with each other, the occurrence of cracking caused by overlarge pressure of input fluid between the flow channels is avoided, and the PDMS is transparent in whole and is easier to observe.
(3) The device is not only suitable for high-pass generation of micro-droplets, but also can generate micro-bubbles in a high-pass manner when a dispersed phase is changed into a gas phase.
Drawings
Fig. 1 is an isometric view of a three-dimensional microfluidic device for high-throughput microdroplet generation according to the present invention.
Fig. 2 (a) is an isometric view of an underlying PDMS flow channel 400, and fig. (b) is a rear view of the underlying PDMS flow channel 400.
Fig. 3 (a) is an isometric view of the middle layer PDMS flow channel 500, and (b) is a rear view of the middle layer PDMS flow channel 500.
Fig. 4 is an isometric view of an upper PDMS flow channel 600.
Fig. 5 is a schematic diagram of micro-droplet generation for a three-dimensional microfluidic device for high-throughput micro-droplet generation.
Detailed Description
The present invention will be described in detail with reference to the accompanying drawings.
Referring to fig. 1, a three-dimensional microfluidic device for high-throughput micro-droplet generation includes a closed flow channel structure formed by bonding a bottom PDMS flow channel 400, a middle PDMS flow channel 500, and an upper PDMS flow channel 600, where the closed flow channel structure is provided with a dispersed phase inlet connector 100, a continuous phase inlet connector 300, and a collection outlet connector 200; the closed micro-channel system is used for accommodating dispersed phase solution and continuous phase solution samples, providing environment for generation of micro-droplets and conveying the generated droplets to a droplet collecting port;
referring to fig. 2, the bottom PDMS flow channel 400 includes two sets of T-shaped droplet generation flow channel arrays, where the first set of T-shaped droplet generation flow channel array includes a first dispersed phase flow channel 402, a first continuous phase flow channel 401, a second continuous phase flow channel 405, and a second dispersed phase flow channel 406, an inlet end of the first dispersed phase flow channel 402 is connected to an inlet end of the second dispersed phase flow channel 406 after meeting and then connected to a first dispersed phase inlet 407, an outlet end of the first dispersed phase flow channel 402 is communicated to a middle portion of the first continuous phase flow channel 401, an inlet end of the first continuous phase flow channel 401 is connected to an inlet end of the second continuous phase flow channel 405 after meeting and then connected to a first continuous phase inlet 414, and an outlet end of the first continuous phase flow channel 401 is connected to a first delivery outlet 403; the outlet end of the second dispersed phase flow channel 406 is communicated with the middle part of the second continuous phase flow channel 405, and the outlet end of the second continuous phase flow channel 405 is connected with the second conveying outlet 404;
the second group of T-shaped micro-droplet generation flow channel array comprises a third dispersed phase flow channel 408, a third continuous phase flow channel 410, a fourth dispersed phase flow channel 412 and a fourth continuous phase flow channel 413, wherein the inlet end of the third dispersed phase flow channel 408 is connected with the first dispersed phase inlet 407 after meeting with the inlet end of the fourth dispersed phase flow channel 412, the outlet end of the third dispersed phase flow channel 408 is communicated with the middle part of the third continuous phase flow channel 410, the inlet end of the third continuous phase flow channel 410 is connected with the first continuous phase inlet 414 after meeting with the inlet end of the fourth continuous phase flow channel 413, and the outlet end of the third continuous phase flow channel 410 is connected with a third conveying outlet 409; the outlet end of the fourth dispersed phase flow channel 412 communicates with the middle of the fourth continuous phase flow channel 413, and the outlet end of the fourth continuous phase flow channel 413 is connected to the fourth delivery outlet 411.
Referring to fig. 3, the middle PDMS flow channel 500 includes two sets of T-shaped micro droplet generation flow channel arrays, where the first set of T-shaped micro droplet generation flow channel array includes a fifth dispersed phase flow channel 503, a fifth continuous phase flow channel 502, a sixth continuous phase flow channel 501 and a sixth dispersed phase flow channel 507, an inlet end of the fifth dispersed phase flow channel 503 is connected to a second dispersed phase inlet 508 after meeting an inlet end of the fifth dispersed phase flow channel 507, an outlet end of the fifth dispersed phase flow channel 503 is connected to a middle portion of the fifth continuous phase flow channel 502, an inlet end of the fifth continuous phase flow channel 502 is connected to a second continuous phase inlet 516 after meeting an inlet end of the sixth continuous phase flow channel 501, and an outlet end of the fifth continuous phase flow channel 502 is connected to a fifth delivery outlet 504; an outlet end of the sixth dispersed phase flow channel 507 is communicated with the middle part of the sixth continuous phase flow channel 501, an outlet end of the sixth continuous phase flow channel 501 is connected with a sixth delivery outlet 507, and a fifth delivery outlet 504 and a sixth delivery outlet 506 are converged at a seventh delivery outlet 505 through the flow channels;
the second group of T-shaped micro droplet generation flow channel arrays includes a seventh dispersed phase flow channel 509, a seventh continuous phase flow channel 515, an eighth continuous phase flow channel 514 and an eighth dispersed phase flow channel 513, an inlet end of the seventh dispersed phase flow channel 509 is connected to the second dispersed phase inlet 508 after meeting with an inlet end of the eighth dispersed phase flow channel 513, an outlet end of the seventh dispersed phase flow channel 509 is communicated with the middle of the seventh continuous phase flow channel 515, an inlet end of the seventh continuous phase flow channel 515 is connected to the second continuous phase inlet 516 after meeting with an inlet end of the eighth continuous phase flow channel 514, and an outlet end of the seventh continuous phase flow channel 515 is connected to the eighth delivery outlet 510; an outlet end of the eighth dispersed phase flow channel 513 is communicated with a middle portion of the eighth continuous phase flow channel 514, an outlet end of the eighth continuous phase flow channel 514 is connected with the ninth delivery outlet 512, and the eighth delivery outlet 510 and the ninth delivery outlet 512 are merged at the tenth delivery outlet 511 through the flow channel.
Referring to fig. 4, the top PDMS flow channel 600 includes a third dispersed phase inlet 604, a third continuous phase inlet 601 and a micro-droplet collecting port 605, where the third dispersed phase inlet 604 is connected to the dispersed phase inlet connector 100, and the third continuous phase inlet 601 is connected to the continuous phase inlet connector 300; the inlet end of the micro-droplet collecting port 605 is connected to the outlet ends of the eleventh and twelfth delivery outlets 603 and 606 through the first and second delivery flow channels 602 and 607.
The liquid drop generating device is characterized by comprising four groups of T-shaped micro-liquid drop generating flow channel arrays so as to realize high-pass generation of liquid drops.
The height of the flow channel of the closed flow channel structure is 60 micrometers, and the height of the flow channel is equal to the height of the flow channel except for a fifth conveying outlet 504, a sixth conveying outlet 506, a seventh conveying outlet 505, an eighth conveying outlet 510, a ninth conveying outlet 512, a second dispersed phase inlet 508 and a tenth conveying outlet 511, wherein a second continuous phase inlet 516, a third dispersed phase inlet 604, a micro-droplet collecting port 605 and a third continuous phase inlet 601 are all through holes, the aperture of the through holes is the same, and all other inlets and outlets are non-through holes.
The closed micro-channel system is made of Polydimethylsiloxane (PDMS) with good light transmittance and biocompatibility, and optical monitoring and recording of the generation process of micro-droplets are facilitated.
The relative position relationship among the bottom layer PDMS flow channel 400, the middle layer PDMS flow channel 500, and the upper layer PDMS flow channel 600 is as follows: the lower surface of the middle PDMS runner 500 without a runner is bonded to the upper surface of the bottom PDMS runner 400 with a runner, and the lower surface of the top PDMS runner 600 is bonded to the upper surface of the middle PDMS runner 500 with a runner; the central axes of the first dispersed phase inlet 407, the second dispersed phase inlet 508, the third dispersed phase inlet 604 and the dispersed phase inlet joint 100 are coaxial and are communicated with each other, so that the dispersed phase can transmit and flow in the vertical direction in the three-dimensional flow channel; the central axes of the first continuous phase inlet 414, the second continuous phase inlet 516, the third continuous phase inlet 601 and the continuous phase inlet connector 300 are coaxial and are communicated with each other, so that the transmission flow of the continuous phase in the vertical direction in the three-dimensional flow channel is realized; the first delivery outlet 403 and the fifth delivery outlet 504, the second delivery outlet 404 and the sixth delivery outlet 506, the third delivery outlet 409 and the eighth delivery outlet 510, the fourth delivery outlet 411 and the ninth delivery outlet 512, the seventh delivery outlet 505 and the eleventh delivery outlet 603, and the tenth delivery outlet 511 and the twelfth delivery outlet 606 are respectively coaxial with each other in central axis, and are communicated with each other to realize the transmission of the generated micro-droplets, and finally delivered to the micro-droplet collection port 605 for collecting the micro-droplets.
The micro-droplet generation method for the three-dimensional micro-fluidic device for high-throughput micro-droplet generation comprises the following steps:
1) fixing the three-dimensional microfluidic device for high-flux micro-droplet generation on an objective table of a microscope, and ensuring that two groups of T-shaped micro-droplet generation flow channel array parts of each middle-layer PDMS flow channel 500 are positioned in a field of view of the microscope and have no inclination through objective observation;
2) respectively connecting a disperse phase inlet connector 100, a continuous phase inlet connector 300 and a collection outlet connector 200 with a disperse phase solution storage bottle, a continuous phase solution storage bottle and a micro-droplet collection container on a nitrogen pressure injection pump through Teflon catheters;
3) starting an injection pump, firstly adjusting the dispersed phase and the continuous phase to corresponding flow rates through the injection pump, continuously shearing the dispersed phase into micro-droplets at the positions of the T-shaped micro-droplet generation flow channel arrays through the fluid shearing force of the continuous phase so as to realize the generation of the micro-droplets, and then conveying the droplets to a droplet collection container through the connectors, the conveying ports, the flow channels and the collection outlet connector.
Referring to fig. 5, the generation process of the micro-droplets in the three-dimensional microfluidic device for high-throughput micro-droplet generation is: the continuous phase solution simultaneously enters a first continuous phase flow channel 401, a second continuous phase flow channel 405, a third continuous phase flow channel 410, a fourth continuous phase flow channel 413, a fifth continuous phase flow channel 502, a sixth continuous phase flow channel 501, a seventh continuous phase flow channel 515 and an eighth continuous phase flow channel 514 through holes of a first continuous phase inlet 414, a second continuous phase inlet 516 and a third continuous phase inlet 601, the dispersed phase solution simultaneously enters a first dispersed phase flow channel 402, a second dispersed phase flow channel 406, a third dispersed phase flow channel 408, a fourth dispersed phase flow channel 412, a fifth dispersed phase flow channel 503, a sixth dispersed phase flow channel 507, a seventh dispersed phase flow channel 509 and an eighth dispersed phase flow channel 604, and the input pressure of the dispersed phase solution and the continuous phase solution is adjusted through a nitrogen pressure injection pump, so that the dispersed phase, the dispersed phase and the dispersed phase are dispersed in a dispersed phase flow channel 406, a dispersed phase flow channel 408, a dispersed phase flow channel 412, a fifth dispersed phase flow channel 503, a, And filling the continuous phase in each flow channel, and adjusting the dispersed phase and the continuous phase to corresponding flow velocities, so that at each T-shaped micro-channel structure, the dispersed phase is continuously sheared into micro-droplets through fluid shearing of the continuous phase and the dispersed phase, and high-flux generation of the micro-droplets is realized.
Claims (7)
1. A three-dimensional microfluidic device for high-throughput microdroplet generation, characterized by: the device comprises a closed flow channel structure formed by bonding a bottom PDMS flow channel (400), a middle PDMS flow channel (500) and an upper PDMS flow channel (600), wherein the closed flow channel structure is provided with a dispersed phase inlet connector (100), a continuous phase inlet connector (300) and a collection outlet connector (200).
2. The three-dimensional microfluidic device for high throughput microdroplet generation of claim 1, wherein: the bottom PDMS flow channel (400) comprises two groups of T-shaped micro-droplet generation flow channel arrays, wherein the first group of T-shaped micro-droplet generation flow channel arrays comprises a first dispersed phase flow channel (402), a first continuous phase flow channel (401), a second continuous phase flow channel (405) and a second dispersed phase flow channel (406), the inlet end of the first dispersed phase flow channel (402) is connected with the first dispersed phase inlet (407) after intersecting with the inlet end of the second dispersed phase flow channel (406), the outlet end of the first dispersed phase flow channel (402) is communicated with the middle part of the first continuous phase flow channel (401), the inlet end of the first continuous phase flow channel (401) is connected with the first continuous phase inlet (414) after intersecting with the inlet end of the second continuous phase flow channel (405), and the outlet end of the first continuous phase flow channel (401) is connected with a first conveying outlet (403); the outlet end of the second dispersed phase flow channel (406) is communicated with the middle part of the second continuous phase flow channel (405), and the outlet end of the second continuous phase flow channel (405) is connected with the second conveying outlet (404);
the second group of T-shaped micro-droplet generation flow channel array comprises a third dispersed phase flow channel (408), a third continuous phase flow channel (410), a fourth dispersed phase flow channel (412) and a fourth continuous phase flow channel (413), wherein the inlet end of the third dispersed phase flow channel (408) is connected with the first dispersed phase inlet (407) after meeting with the inlet end of the fourth dispersed phase flow channel (412), the outlet end of the third dispersed phase flow channel (408) is communicated with the middle part of the third continuous phase flow channel (410), the inlet end of the third continuous phase flow channel (410) is connected with the first continuous phase inlet (414) after meeting with the inlet end of the fourth continuous phase flow channel (413), and the outlet end of the third continuous phase flow channel (410) is connected with a third conveying outlet (409); the outlet end of the fourth dispersed phase flow channel (412) is communicated with the middle part of the fourth continuous phase flow channel (413), and the outlet end of the fourth continuous phase flow channel (413) is connected with the fourth conveying outlet (411).
3. The three-dimensional microfluidic device for high throughput microdroplet generation of claim 1, wherein: the middle PDMS flow channel (500) comprises two groups of T-shaped micro-droplet generation flow channel arrays, wherein the first group of T-shaped micro-droplet generation flow channel array comprises a fifth dispersed phase flow channel (503), a fifth continuous phase flow channel (502), a sixth continuous phase flow channel (501) and a sixth dispersed phase flow channel (507), the inlet end of the fifth dispersed phase flow channel (503) is connected with the second dispersed phase inlet (508) after meeting with the inlet end of the fifth dispersed phase flow channel (507), the outlet end of the fifth dispersed phase flow channel (503) is communicated with the middle part of the fifth continuous phase flow channel (502), the inlet end of the fifth continuous phase flow channel (502) is connected with the second continuous phase inlet (516) after meeting with the inlet end of the sixth continuous phase flow channel (501), and the outlet end of the fifth continuous phase flow channel (502) is connected with a fifth conveying outlet (504); the outlet end of the sixth dispersed phase flow channel (507) is communicated with the middle part of the sixth continuous phase flow channel (501), the outlet end of the sixth continuous phase flow channel (501) is connected with a sixth conveying outlet (507), and a fifth conveying outlet (504) and a sixth conveying outlet (506) are converged at a seventh conveying outlet (505) through the flow channels;
the second group of T-shaped micro-droplet generation flow channel array comprises a seventh dispersed phase flow channel (509), a seventh continuous phase flow channel (515), an eighth continuous phase flow channel (514) and an eighth dispersed phase flow channel (513), wherein the inlet end of the seventh dispersed phase flow channel (509) is connected with the second dispersed phase inlet (508) after meeting with the inlet end of the eighth dispersed phase flow channel (513), the outlet end of the seventh dispersed phase flow channel (509) is communicated with the middle part of the seventh continuous phase flow channel (515), the inlet end of the seventh continuous phase flow channel (515) is connected with the second continuous phase inlet (516) after meeting with the inlet end of the eighth continuous phase flow channel (514), and the outlet end of the seventh continuous phase flow channel (515) is connected with the eighth delivery outlet (510); the outlet end of the eighth dispersed phase flow channel (513) is communicated with the middle part of the eighth continuous phase flow channel (514), the outlet end of the eighth continuous phase flow channel (514) is connected with the ninth delivery outlet (512), and the eighth delivery outlet (510) and the ninth delivery outlet (512) are converged at the tenth delivery outlet (511) through the flow channels.
4. The three-dimensional microfluidic device for high throughput microdroplet generation of claim 1, wherein: the top PDMS flow channel (600) comprises a third dispersed phase inlet (604), a third continuous phase inlet (601) and a micro-droplet collection port (605), wherein the third dispersed phase inlet (604) is connected with a dispersed phase inlet joint (100), and the third continuous phase inlet (601) is connected with a continuous phase inlet joint (300); the inlet end of the micro-droplet collecting port (605) is connected with the outlet ends of the eleventh delivery outlet (603) and the twelfth delivery outlet (606) through the first delivery flow channel (602) and the second delivery flow channel (607).
5. The three-dimensional microfluidic device for high throughput micro-droplet generation of claim 2, 3 or 4, wherein: the height of the flow channel of the closed flow channel structure is 60 micrometers, except for a fifth conveying outlet (504), a sixth conveying outlet (506), a seventh conveying outlet (505), an eighth conveying outlet (510), a ninth conveying outlet (512), a second dispersed phase inlet (508) and a tenth conveying outlet (511), the second continuous phase inlet (516), the third dispersed phase inlet (604), the micro-droplet collecting port (605) and the third continuous phase inlet (601) are all through holes, the diameters of the through holes are the same, and the other inlets and outlets are all non-through holes, and the heights of the inlets and outlets are equal to the height of the flow channel.
6. The three-dimensional microfluidic device for high throughput micro-droplet generation of claim 2, 3 or 4, wherein: the relative position relationship among the bottom layer PDMS flow channel (400), the middle layer PDMS flow channel (500) and the upper layer PDMS flow channel (600) is as follows: the lower surface of the middle-layer PDMS runner (500) without the runner is bonded to the upper surface of the bottom-layer PDMS runner (400) with the runner, and the lower surface of the top-layer PDMS runner (600) is bonded to the upper surface of the middle-layer PDMS runner (500) with the runner; the central axes of the first dispersed phase inlet (407), the second dispersed phase inlet (508), the third dispersed phase inlet (604) and the dispersed phase inlet joint (100) are coaxial and are communicated with each other; the central axes of the first continuous phase inlet (414), the second continuous phase inlet (516), the third continuous phase inlet (601) and the continuous phase inlet joint (300) are coaxial and are communicated with each other; the first conveying outlet (403) and the fifth conveying outlet (504), the second conveying outlet (404) and the sixth conveying outlet (506), the third conveying outlet (409) and the eighth conveying outlet (510), the fourth conveying outlet (411) and the ninth conveying outlet (512), the seventh conveying outlet (505) and the eleventh conveying outlet (603), and the tenth conveying outlet (511) and the twelfth conveying outlet (606) are coaxial in central axis and mutually communicated.
7. A method of generating micro-droplets for a three-dimensional microfluidic device for high throughput micro-droplet generation according to claim 1, comprising the steps of:
1) fixing a three-dimensional microfluidic device for high-flux micro-droplet generation on an objective table of a microscope, and ensuring that two groups of T-shaped micro-droplet generation flow channel array parts of each middle-layer PDMS flow channel (500) are positioned in a microscope field of view and are not inclined through objective observation;
2) respectively connecting a disperse phase inlet connector (100), a continuous phase inlet connector (300) and a collection outlet connector (200) with a disperse phase solution storage bottle, a continuous phase solution storage bottle and a micro-droplet collection container on a nitrogen pressure injection pump through Teflon catheters;
3) and starting the injection pump, adjusting the dispersed phase and the continuous phase to corresponding flow rates through the injection pump, and shearing the dispersed phase into micro droplets by the continuous phase at the positions where the T-shaped micro droplets generate the flow channel array.
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