Microfluidic chip, device for high-throughput preparation of microspheres, and microsphere preparation method thereof

Abstract

A microfluidic chip, a device for high-throughput preparation of microspheres, and a microsphere preparation method thereof. The microfluidic chip (3) comprises a mixed phase outlet (36), and a plurality of microsphere preparation channels communicated with the mixed phase outlet (36) and arranged in parallel with each other. Each microsphere preparation channel comprises a continuous phase channel (31), a disperse phase channel (32) and a flow focusing structure (33), wherein the continuous phase channel (31) allows a continuous phase inlet (34) to be communicated with the flow focusing structure (33), the disperse phase channel (32) allows a disperse phase inlet (35) to be communicated with the flow focusing structure (33), and the flow focusing structure (33) is communicated with the mixed phase outlet (36). The flow focusing structures (33) are configured to disperse a disperse phase fluid into a plurality of microspheres by means of a flow shear force effect of a main phase fluid in the continuous phase channels (31) on the dispersed phase fluid in the disperse phase channels (32). The device for high-throughput preparation of microspheres comprises an interface assembly (2), a first pump member, a second pump member and the microfluidic chip (3). The present invention can provide the high-throughput preparation performance, so as to meet the requirements of fields such as biology and chemistry for a large quantity of microspheres.

Description

Microfluidic chip, device for high-throughput microsphere preparation, and method for preparing microspheres

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application number CN202311062532.3 and filing date August 22, 2023, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present invention belongs to the field of microfluidic microsphere preparation, and particularly relates to a microfluidic chip, a high-throughput microsphere preparation device and a microsphere preparation method thereof. Background Art

[0004] Microfluidics is an emerging discipline, driven primarily by advances in micro- and nanoscale processing and the urgent need for new materials in biochemistry. It enables the design and manipulation of chemical and biological processes at the microscale, demonstrating tremendous potential in addressing obstacles that traditional chemical and biological methods cannot address. Early on, microfluidics, also known as the micro-total analysis laboratory, integrated some or all laboratory functions onto a single chip for sample separation, mixing, and detection. With advances in design processes, micro- and nanofabrication, and labeling, microfluidics has expanded beyond micro-total analysis to encompass a wide range of disciplines, including biochemistry and physiology. In particular, after more than two decades of rapid development, microfluidics has become a vital tool in the fields of biology, chemical engineering, and pharmaceuticals. It serves as an alternative to existing technologies, such as cell culture, emulsification, and drug encapsulation, and also as a novel technology offering new opportunities for industrial efficiency, such as in microreactors, fine chemicals, and point-of-care (POCT) chips.

[0005] A typical application of microfluidics is to prepare microspheres. By using a continuous phase shear fluid to overcome surface tension and discretize it, small microspheres are formed. In the presence of surfactants, the microspheres can exist stably. Microspheres have a variety of applications. They can be used as microreactors to control various biochemical reactions inside microscale microspheres; they can be used to encapsulate cells and provide the necessities for cell activity; they can also be used for targeted drug delivery, by modifying chemokines on the surface of microspheres to deliver drugs in a targeted manner to the target area. The preparation methods of microspheres can include T-channel method, flow focusing method, confocal method, etc. according to the preparation structure, but the basic principle is to use the shear effect of a continuous phase on another phase to form discrete microspheres.

[0006] Microspheres can be either water-in-oil or oil-in-water, with the continuous and discrete phases configured according to the specific application scenario. Water-in-oil microspheres are commonly used, and the aqueous solution in water microspheres can be either a water solvent solution or a hydrogel solution. In the biomedical field, hydrogel solutions are the most common. Hydrogels are biocompatible materials, with over 80% of their volume consisting of free water, thus providing a viable environment for cells and other biological elements. Because hydrogels can be cross-linked using various methods and degrees of cross-linking, they offer a wide range of mechanical properties and are used to prepare various biomaterials, such as drug capsules, microtissues, and culture media. Commonly used hydrogels include PEG-based hydrogels, PLAG hydrogels, dextran, and alginate hydrogels. Different hydrogel systems can be designed based on requirements such as biodegradability, temperature sensitivity, and pH responsiveness, depending on the application scenario.

[0007] Since microfluidic chips can provide size-controlled aqueous microspheres at high throughput, they can be widely used in the preparation of hydrogel microspheres. However, the current preparation methods are mostly single-channel, with frequencies ranging from a few hundred to a few thousand hertz, while the microspheres used in the biological field require quantities ranging from millions to billions. In addition, the characteristic size of microspheres that can be generated by the channels of current microsphere preparation chips is mostly above 15 microns, while the microspheres used for bionic needs require their characteristic size to be consistent with the common cell size, that is, ranging from 5 microns to 20 microns. In summary, it is necessary to design and manufacture a high-throughput aqueous microsphere preparation chip that can meet the preparation needs of high-throughput microspheres and further reduce the size of the microspheres.

[0008] Summary of the Invention

[0009] In view of the shortcomings of the existing technology, the problem of low throughput of microsphere preparation and too high lower limit of microsphere diameter adjustment is solved, so as to meet the high throughput preparation of microspheres while providing a wider range of microsphere diameter adjustment.

[0010] According to a first aspect of the present invention, there is provided a microfluidic chip, comprising a mixed phase outlet and a plurality of microsphere preparation channels connected to the mixed phase outlet and arranged in parallel with each other;

[0011] Each of the microsphere preparation channels comprises a continuous phase channel, a discrete phase channel and a flow focusing structure, wherein the continuous phase channel connects the continuous phase inlet with the flow focusing structure, the discrete phase channel connects the discrete phase inlet with the flow focusing structure, and the flow focusing structure connects the mixed phase outlet;

[0012] The flow focusing structure is configured to disperse the dispersed phase fluid into a plurality of microspheres through the flow shear force of the main phase fluid in the continuous phase channel on the dispersed phase fluid in the discrete phase channel.

[0013] Preferably, the continuous phase channel includes a continuous phase Y-shaped diversion section, a first continuous phase diversion section and a second continuous phase diversion section;

[0014] The continuous phase inlet is connected to the inlet of the continuous phase Y-shaped diverter section, the two outlets of the continuous phase Y-shaped diverter section are connected to the first continuous phase diverter section and the second continuous phase diverter section respectively, and the first continuous phase diverter section and the second continuous phase diverter section are both connected to the flow focusing structure;

[0015] The connection position between the first continuous phase diversion section and the flow focusing structure and the connection position between the second continuous phase diversion section and the flow focusing structure are respectively placed on both sides of the connection position between the discrete phase channel and the flow focusing structure.

[0016] Preferably, the flow focusing structure is in a cross shape.

[0017] Preferably, the discrete phase channel includes a first discrete phase segment, a second discrete phase segment and a discrete phase Y-shaped mixing segment, and the discrete phase inlet includes a first discrete phase inlet and a second discrete phase inlet;

[0018] The first discrete phase segment connects the first discrete phase inlet with one inlet of the discrete phase Y-shaped mixing segment, the second discrete phase segment connects the second discrete phase inlet with the other inlet of the discrete phase Y-shaped mixing segment, and the outlet of the discrete phase Y-shaped mixing segment is connected to the flow focusing structure.

[0019] Preferably, the downstream side of the flow focusing structure has a gradually expanding section, and the width of the gradually expanding section gradually increases along the flow direction of the liquid.

[0020] Preferably, the number of the microsphere preparation channels is an even number, all the microsphere preparation channels are grouped in pairs, and the two microsphere preparation channels in each group are connected to the mixed phase outlet through the same incorporation structure.

[0021] Preferably, the two microsphere preparation channels in each group are arranged adjacent to each other.

[0022] Preferably, each of the merging structures is connected to the mixed phase outlet via an inflow channel, and each of the inflow channels has at least one bend section.

[0023] Preferably, all the inflow channels have the same number of curved sections.

[0024] Preferably, the outlets of all the inflow channels are evenly arranged around the circumference of the mixed phase outlet.

[0025] Preferably, the flow focusing structure has a first continuous phase inflow section, a second continuous phase inflow section, a discrete phase inflow section and an outflow section, the first continuous phase inflow section is connected to the first continuous phase diversion section, the second continuous phase inflow section is connected to the second continuous phase diversion section, the discrete phase inflow section is connected to the discrete phase channel, and the first continuous phase inflow section, the second continuous phase inflow section and the discrete phase inflow section have the same width and the width is d.

[0026] Preferably, the length of the outflow section is 3d-6d; and / or,

[0027] The arc radius of the gradually expanding section is 4d; and / or,

[0028] The length of the gradually expanding section is 10d-15d; and / or,

[0029] The width of the downstream side of the gradually expanding section is 4d-6d.

[0030] Preferably, the continuous phase inlet is connected to the inlet of the continuous phase Y-shaped diversion section through a continuously connected section, the width of the continuously connected section is 3.2d, the first section width of the first continuous phase diversion section is 1.6d, the tail section is tilted and the width gradually shrinks from 1.6d to d, the first section width of the second continuous phase diversion section is 1.6d, the tail section is tilted and the width gradually shrinks from 1.6d to d.

[0031] Preferably, the value range of d is 30-60 microns.

[0032] Preferably, d is 50 microns.

[0033] Preferably, the microfluidic chip is made of stainless steel, COC, PMMA, PC, PE or glass.

[0034] According to a second aspect of the present invention, there is provided a device for preparing microspheres with high throughput, the device comprising an interface assembly, a first pump component, a second pump component, and the microfluidic chip described in any one of the above;

[0035] The interface component has a continuous phase interface, a discrete phase interface and a mixed phase interface. The continuous phase interface is connected to the first pump component and the continuous phase inlet, the discrete phase interface is connected to the second pump component and the discrete phase inlet, and the mixed phase interface is connected to the mixed phase outlet.

[0036] Preferably, the interface component includes an inlet mechanism and an outlet mechanism, the continuous phase interface and the discrete phase interface are both arranged on the inlet mechanism, the mixed phase interface is arranged on the outlet mechanism, the part of the microfluidic chip having the continuous phase inlet and the discrete phase inlet is arranged on the inlet mechanism, and the part of the microfluidic chip having the mixed phase outlet is arranged on the outlet mechanism.

[0037] Preferably, the inlet mechanism includes a connected inlet upper plate and an inlet lower plate, the part of the microfluidic chip having the continuous phase inlet and the discrete phase inlet is clamped between the inlet upper plate and the inlet lower plate, and the continuous phase interface and the discrete phase interface are both arranged on the inlet upper plate.

[0038] Preferably, an inlet gasket is provided between the inlet upper plate and the microfluidic chip, and a continuous phase buffer groove and a discrete phase buffer groove are formed on the inlet gasket. The continuous phase buffer groove connects the continuous phase interface with the continuous phase inlet, and the discrete phase buffer groove connects the discrete phase interface with the discrete phase inlet.

[0039] Preferably, a first gasket receiving groove is formed on the inlet upper plate, and a portion of the inlet gasket is received in the first gasket receiving groove.

[0040] Preferably, the first gasket receiving groove has at least one positioning protrusion, and the inlet gasket is formed with positioning holes that correspond one-to-one with the positioning protrusion and are positioned and matched with each other.

[0041] Preferably, the inlet upper plate and the inlet lower plate are connected by a plurality of fastening bolts, so that the inlet gasket is sealed and fitted with the inlet upper plate and the microfluidic chip respectively.

[0042] Preferably, the inlet upper plate is provided with at least one positioning block, and the inlet lower plate is provided with a positioning groove which corresponds one-to-one with and is positioned to fit with the positioning block.

[0043] Preferably, the outlet mechanism includes a connected outlet upper plate and an outlet lower plate, the portion of the microfluidic chip having the mixed phase outlet is sandwiched between the outlet upper plate and the outlet lower plate, and the mixed phase interface is arranged on the outlet upper plate.

[0044] Preferably, an outlet gasket is provided between the outlet upper plate and the microfluidic chip, and a mixed phase opening is formed on the outlet gasket, and the mixed phase opening connects the mixed phase interface with the mixed phase outlet.

[0045] Preferably, a second gasket receiving groove is formed on the outlet upper plate, and a portion of the outlet gasket is received in the second gasket receiving groove.

[0046] Preferably, the outlet upper plate and the outlet lower plate are connected by a plurality of fastening bolts, so that the outlet gasket is sealed and fitted with the outlet upper plate and the microfluidic chip respectively.

[0047] Preferably, the shapes of the continuous phase inlet and the discrete phase inlet are both circular holes, all the continuous phase inlets are evenly arranged in parallel, and all the discrete phase inlets are evenly arranged in parallel.

[0048] According to a third aspect of the present invention, there is provided a method for preparing microspheres based on any one of the above-described high-throughput microsphere preparation devices, the method comprising the following steps:

[0049] S1: dissolving a cross-linkable synthetic biomaterial and / or a cross-linkable natural biomaterial in an aqueous solvent and filtering the solution through a 0.22 μm filter membrane to form a continuous aqueous phase solution;

[0050] S2: The organic oil phase reagent and the surfactant are thoroughly mixed in a mass ratio of 3:1-5:1, and filtered through a 0.22 μm filter membrane to form a discrete phase oil phase solution;

[0051] S3: mixing the crosslinker into the oil phase as the collection liquid;

[0052] S4: pumping the continuous phase aqueous solution into the continuous phase interface through the first pump and controlling the aqueous phase flow rate to be 0.2-30 μL / min, and pumping the dispersed phase oil phase solution into the dispersed phase interface through the second pump and controlling the oil phase flow rate to be 20-1000 μL / min;

[0053] S5: receiving the formed microspheres into the collection liquid through the mixed phase interface;

[0054] S6: Separate the microspheres by filtration, washing and centrifugation.

[0055] Preferably, the cross-linkable artificial synthetic biomaterial is selected from one or more of polyanhydrides, polyesters, polyamides, polyamino acids, polypropylene, polystyrene, polyacrylamide, polyethylene glycol, polyethylene glycol derivatives, polylactic acid, polyhydroxy acid, polylactic acid-alcohol copolymers, polypyrrole, polyester, polymethacrylate, polydimethylsiloxane, polyacetal, polycyanoacrylate, polyurethane, polyethylene, polycarbonate or polyethylene oxide.

[0056] Preferably, the cross-linkable natural biomaterial is selected from one or more of gelatin, gelatin derivatives, alginate, alginate derivatives, collagen, proteoglycan, glycoprotein, agar, matrigel, hyaluronic acid, laminin, fibronectin, chitosan, chitosan derivatives, and soy protein.

[0057] Preferably, the organic oil phase reagent is selected from one or more of mineral oil, long-chain alkanes, and HFE7500 fluorinated oil.

[0058] Preferably, the surfactant is Span 80 or Tween 80.

[0059] Preferably, the cross-linking agent is selected from one or more of glutaraldehyde, genipin, transglutaminase, tetramethylethylenediamine, ammonium persulfate, calcium sulfate, calcium chloride, zinc sulfate, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

[0060] Preferably, the aqueous phase flow rate is 7 μL / min; and / or,

[0061] The oil phase flow rate was 40 μL / min.

[0062] Preferably, in step S4, the continuous phase aqueous solution is a continuous phase aqueous solution after air bubbles are exhausted, and the dispersed phase oil phase solution is a dispersed phase oil phase solution after air bubbles are exhausted.

[0063] The present invention can be used for the preparation of microspheres and gel microspheres. The flow focusing method is used to prepare microspheres, which can reduce the lower limit of microsphere diameter adjustment while providing a wider microsphere diameter adjustment range. The parallel arrangement method is used to integrate multiple focusing structures on a chip, which can provide high-throughput preparation performance and meet the demand for large-scale microspheres in fields such as biology and chemistry. BRIEF DESCRIPTION OF THE DRAWINGS

[0064] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the following description of the embodiments with reference to the accompanying drawings, in which:

[0065] FIG1 is a schematic diagram of the overall structure of a high-throughput microsphere preparation device of the present invention;

[0066] FIG2 is an exploded view of a high-throughput microsphere preparation device of the present invention;

[0067] FIG3 is a first structural diagram of an inlet upper plate in an inlet mechanism of an interface assembly of a high-throughput microsphere preparation device of the present invention;

[0068] FIG4 is a second structural diagram of the inlet upper plate in the inlet mechanism of the interface assembly of the apparatus for high-throughput microsphere preparation of the present invention;

[0069] FIG5 is a first structural diagram of an outlet upper plate in an outlet mechanism of an interface assembly of a high-throughput microsphere preparation device of the present invention;

[0070] FIG6 is a second structural diagram of the outlet upper plate in the outlet mechanism of the interface assembly of the apparatus for high-throughput microsphere preparation of the present invention;

[0071] FIG7 is a schematic diagram of the overall structure of the microfluidic chip of the present invention;

[0072] FIG8 is a partial schematic diagram of the microfluidic chip of the present invention;

[0073] FIG9 is a schematic structural diagram of an inlet gasket of a high-throughput microsphere preparation device of the present invention;

[0074] FIG10 is an optical microscopic image of a plurality of microspheres produced by the high-throughput microsphere production apparatus of the present invention;

[0075] FIG11 is an electron micrograph of a plurality of microspheres produced by the high-throughput microsphere production apparatus of the present invention;

[0076] FIG12 is an electron microscope image of a single microsphere prepared by the high-throughput microsphere preparation device of the present invention.

[0077] List of Reference Numerals: 1. Apparatus for high-throughput microsphere preparation; 2. Interface assembly; 2a. Continuous phase interface; 2b. Discrete phase interface; 2c. Mixed phase interface; 21. Inlet mechanism; 211. Inlet upper plate; 211a. First gasket receiving groove; 211b. Positioning protrusion; 211c. Positioning block; 212. Inlet lower plate; 212c. Positioning groove; 22. Outlet mechanism; 221. Outlet upper plate; 221a. Second gasket receiving groove; 222. Outlet lower plate; 3. Microfluidic chip; 31. Continuous phase channel; 31a. Continuous phase Y-shaped diversion section; 31b. First continuous phase diversion section; 31c. Second continuous phase diversion section; 31d. Continuous-phase connecting section; 32. Discrete phase channel; 32a. First discrete phase section; 32b. Second discrete phase section; 32c. Discrete phase Y-shaped mixing section; 33. Flow focusing structure; 33a. First continuous phase inflow section; 33b. Second continuous phase inflow section; 33c. Dispersed phase inflow section; 33d. Outflow section; 34. Continuous phase inlet; 35. Dispersed phase inlet; 35a. First discrete phase inlet; 35b. Second discrete phase inlet; 36. Mixed phase outlet; 37. Gradual expansion section; 38. Incorporation structure; 39. Inflow channel; 4. Inlet gasket; 4a. Continuous phase buffer tank; 4b. Dispersed phase buffer tank; 4c. Positioning hole; 5. Outlet gasket. DETAILED DESCRIPTION

[0078] The embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and intended to be used to explain the present invention, but should not be understood as limiting the present invention.

[0079] Some terms in this document are explained and illustrated below. Those skilled in the art should understand that these explanations and illustrations are only used to facilitate understanding and should not be regarded as limiting the scope of protection of this application.

[0080] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like to indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention 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, and therefore should not be understood as limiting the present invention.

[0081] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of the technical features being referred to. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "plurality" means at least two, such as two, three, etc., unless otherwise specifically defined.

[0082] In the present invention, unless otherwise specified or limited, the terms "installed," "connected," "connect," "fixed," etc. should be understood in a broad sense. For example, they can refer to fixed connection, detachable connection, or integration; mechanical connection, electrical connection, or communication; direct connection or indirect connection through an intermediate medium; internal communication between two elements or interaction between two elements, unless otherwise specified. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0083] In the present invention, unless otherwise expressly specified or limited, when a first feature is "above" or "below" a second feature, it may mean that the first and second features are in direct contact, or that the first and second features are in indirect contact through an intermediary. Furthermore, when a first feature is "above," "above," or "above" a second feature, it may mean that the first feature is directly above or diagonally above the second feature, or simply means that the first feature is at a higher level than the second feature. When a first feature is "below," "below," or "below" a second feature, it may mean that the first feature is directly below or diagonally below the second feature, or simply means that the first feature is at a lower level than the second feature.

[0084] In the present invention, the terms "one embodiment", "some embodiments", "examples", "specific examples", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic expressions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and the features of different embodiments or examples without contradiction.

[0085] As shown in Figures 1 to 7, the high-throughput microsphere preparation device 1 of the present invention includes an interface component 2, a first pump component (not shown in the figure), a second pump component (not shown in the figure) and a microfluidic chip 3. The interface component 2 has a continuous phase interface 2a, a discrete phase interface 2b and a mixed phase interface 2c. The microfluidic chip 3 includes a mixed phase outlet 36 and a plurality of microsphere preparation channels that are connected to the mixed phase outlet 36 and are arranged in parallel with each other. The flow resistance is controlled to ensure that the flow rate and pressure of each microsphere preparation channel are consistent. Each microsphere preparation channel includes a continuous phase channel 31, a discrete phase channel 32 and a flow focusing structure 33. The continuous phase channel 31 connects the continuous phase inlet 34 with the flow focusing structure 33. The discrete phase channel 32 connects the discrete phase inlet 35 with the flow focusing structure 33, and the flow focusing structure 33 is connected with the mixed phase outlet 36; the flow focusing structure 33 is configured to disperse the dispersed phase fluid into a plurality of microspheres through the flow shear force of the main phase fluid in the continuous phase channel 31 on the dispersed phase fluid in the discrete phase channel 32, the continuous phase interface 2a is connected to the first pump component and connected to the continuous phase inlet 34, the discrete phase interface 2b is connected to the second pump component and connected to the discrete phase inlet 35, and the mixed phase interface 2c is connected to the mixed phase outlet 36, wherein the first pump component and the second pump component preferably adopt microinjection pumps. Of course, in actual applications, other pump components that meet the needs can also be selected. The advantage of this arrangement is that each interface corresponds to a power source, saving costs.

[0086] In some embodiments, referring to Figures 1 to 3 and 5, the continuous phase interface 2a, the discrete phase interface 2b, and the mixed phase interface 2c all have a sinking mechanism. The sinking structure can be formed on the interface boss. The sinking structure is formed with a threaded interface, which can be connected to a threaded pipe. The threaded pipe is then connected to the corresponding microinjection pump. Exemplarily, the microinjection pump is connected to the threaded interface via the threaded pipe, thereby achieving communication between the microinjection pump and the continuous phase interface 2a. In actual applications, the connection between the pipe and the interface can be achieved through a Luer connector to ensure sealing. The threaded interface can be selected in a specification of 1 / 4-28 UNF. In other examples, the interface can also be connected to the corresponding pipeline by a snap-on connection.

[0087] In one embodiment, referring to Figures 1 and 2, the interface assembly 2 includes an inlet mechanism 21 and an outlet mechanism 22. The continuous phase interface 2a and the discrete phase interface 2b are both provided on the inlet mechanism 21, and the mixed phase interface 2c is provided on the outlet mechanism 22. The portion of the microfluidic chip 3 having the continuous phase inlet 34 and the discrete phase inlet 35 is provided on the inlet mechanism 21, and the portion of the microfluidic chip 3 having the mixed phase outlet 36 is provided on the outlet mechanism 22. In a specific example, the inlet mechanism 21 and the outlet mechanism 22 are designed to be split, exposing the flow focusing structure 33 of the microfluidic chip 3 between the inlet mechanism 21 and the outlet mechanism 22, thereby reducing assembly difficulty, saving materials, and lowering material costs.

[0088] Preferably, referring to Figures 1 to 4, 7 and 9, the inlet mechanism 21 includes a connected inlet upper plate 211 and an inlet lower plate 212. The portion of the microfluidic chip 3 having the continuous phase inlet 34 and the discrete phase inlet 35 is sandwiched between the inlet upper plate 211 and the inlet lower plate 212. The continuous phase interface 2a and the discrete phase interface 2b are both provided on the inlet upper plate 211. Further preferably, an inlet gasket 4 is provided between the inlet upper plate 211 and the microfluidic chip 3. The inlet gasket 4 is formed with a continuous phase buffer groove 4a and a discrete phase buffer groove 4b. The continuous phase buffer groove 4a connects the continuous phase interface 2a with the continuous phase inlet 34, and the discrete phase buffer groove 4b connects the discrete phase interface 2b with the discrete phase inlet 35. The continuous phase buffer groove 4a can buffer the main phase fluid flowing from the continuous phase interface 2a to the continuous phase inlet 34, and the discrete phase buffer groove 4b can buffer the dispersed phase fluid flowing from the discrete phase interface 2b to the discrete phase inlet 35.

[0089] Those skilled in the art can flexibly set the specific connection method of the inlet upper plate 211 and the inlet lower plate 212. In a specific example, the inlet upper plate 211 and the inlet lower plate 212 are connected by multiple fastening bolts (as shown in Figures 1 to 3, they can be connected by 4 fastening bolts), so that the inlet gasket 4 is sealed and fitted with the inlet upper plate 211 and the microfluidic chip 3 respectively, and is convenient for disassembly and cleaning. Preferably, a first gasket receiving groove 211a is formed on the inlet upper plate 211, and a part of the inlet gasket 4 is received in the first gasket receiving groove 211a. The inlet gasket 4 can be limited to a certain extent by the first gasket receiving groove 211a. Further preferably, there is at least one positioning protrusion 211b in the first gasket receiving groove 211a, and a positioning hole 4c corresponding to the positioning protrusion 211b and positioned and matched with each other is formed on the inlet gasket 4. As shown in Figures 2 and 4, two positioning protrusions 211b are positioned and matched with two positioning holes 4c.

[0090] In one example, at least one positioning block 211c is provided on the inlet upper plate 211, and a positioning groove 212c corresponding to and positioned with the positioning block 211c is provided on the inlet lower plate 212. As shown in Figures 2 and 4, two positioning blocks 211c are positioned and matched with two positioning grooves 212c in a one-to-one manner.

[0091] Preferably, referring to Figures 1, 2, and 5 to 7, the outlet mechanism 22 includes a connected upper outlet plate 221 and a lower outlet plate 222. The portion of the microfluidic chip 3 having the mixed-phase outlet 36 is sandwiched between the upper outlet plate 221 and the lower outlet plate 222, and the mixed-phase interface 2c is provided on the upper outlet plate 221. Further preferably, an outlet gasket 5 is provided between the upper outlet plate 221 and the microfluidic chip 3. A mixed-phase opening is formed on the outlet gasket 5. The mixed-phase opening connects the mixed-phase interface 2c with the mixed-phase outlet 36. The mixed-phase opening ensures that liquid flowing into the mixed-phase outlet 36 flows to the mixed-phase interface 2c.

[0092] Those skilled in the art can flexibly set the specific connection method of the outlet upper plate 221 and the outlet lower plate 222. In a specific example, the outlet upper plate 221 and the outlet lower plate 222 are connected by multiple fastening bolts (as shown in Figures 1, 2 and 5, they can be connected by two fastening bolts) so that the outlet gasket 5 is sealed and fitted with the outlet upper plate 221 and the microfluidic chip 3 respectively, and is easy to disassemble and clean. Preferably, a second gasket receiving groove 221a is formed on the outlet upper plate 221, and a part of the outlet gasket 5 is received in the second gasket receiving groove 221a. The outlet gasket 5 can be limited to a certain extent by the second gasket receiving groove 221a. Further preferably, the shape of the second gasket receiving groove 221a and the shape of the outlet gasket 5 are both circular.

[0093] In the above embodiment, the inlet gasket 4 and the outlet gasket 5 can be made of perfluoroelastomer (FFKM) or fluoroelastomer (FKM), which have good corrosion resistance and can provide long-lasting sealing. The microfluidic chip 3 can be made of stainless steel, COC, PMMA, PC, PE, glass, etc., and can be processed by photolithography, injection molding, laser processing, etc.

[0094] Preferably, as shown in Figures 1 to 3 and 7, the continuous phase interface 2a is circular in shape, and a plurality of continuous phase inlets 34 are evenly arranged in parallel on the microfluidic chip 3. The continuous phase interface 2a is connected to all the continuous phase inlets 34, and the shape of all the continuous phase inlets 34 is circular. Those skilled in the art can flexibly set the number of continuous phase inlets 34 in actual applications, for example, 4 to 16. In the example of the invention, the number of continuous phase inlets 34 is 8.

[0095] The preferred embodiment of the microfluidic chip 3 of the present invention is further explained below. Referring to Figures 7 and 8, the continuous phase channel 31 includes a continuous phase Y-shaped diversion section 31a, a first continuous phase diversion section 31b and a second continuous phase diversion section 31c; the continuous phase inlet 34 is connected to the inlet of the continuous phase Y-shaped diversion section 31a, and the two outlets of the continuous phase Y-shaped diversion section 31a are respectively connected to the first continuous phase diversion section 31b and the second continuous phase diversion section 31c, and the first continuous phase diversion section 31b and the second continuous phase diversion section 31c are both connected to the flow focusing structure 33; the connection position of the first continuous phase diversion section 31b and the flow focusing structure 33 and the connection position of the second continuous phase diversion section 31c and the flow focusing structure 33 are respectively placed on both sides of the connection position of the discrete phase channel 32 and the flow focusing structure 33. Further preferably, the continuous phase inlet 34 is connected to the inlet of the continuous phase Y-shaped diversion section 31a through the continuous connecting section 31d. Through the continuous phase Y-shaped diversion section 31a, the main phase fluid flowing into the continuous phase inlet 34 can be diverted into the first continuous phase diversion section 31b and the second continuous phase diversion section 31c, so that the dispersed phase fluid forms a plurality of microspheres through the shear force of the fluid on both sides of the main phase fluid.

[0096] Preferably, the flow focusing structure 33 is shaped like a cross. In a specific example, referring to FIG8 , the flow focusing structure 33 comprises a first continuous phase inflow section 33a, a second continuous phase inflow section 33b, a discrete phase inflow section 33c, and an outflow section 33d. The discrete phase inflow section 33c and the outflow section 33d are collinear. The first continuous phase inflow section 33a and the second continuous phase inflow section 33b are collinear. The straight line on which the first continuous phase inflow section 33a and the second continuous phase inflow section 33b lie is perpendicular to the straight line on which the discrete phase inflow section 33c and the outflow section 33d lie. The first continuous phase inflow section 33a is connected to the first continuous phase diversion section 33b, the second continuous phase inflow section 33a is connected to the second continuous phase diversion section 33b, the discrete phase inflow section 33c is connected to the discrete phase channel 32, and the first continuous phase inflow section 33a, the second continuous phase inflow section 33b, and the discrete phase inflow section 33c have the same width, which is d. The value range of d is 30-60 microns, preferably 50 microns, so that the diameter of the prepared microspheres can be adjusted within the range of 5-80 microns.

[0097] Preferably, referring to Figures 7 and 8, the discrete phase channel 32 includes a first discrete phase segment 32a, a second discrete phase segment 32b, and a discrete phase Y-shaped mixing segment 32c, and the discrete phase inlet 35 includes a first discrete phase inlet 35a and a second discrete phase inlet 35b; the first discrete phase segment 32a connects the first discrete phase inlet 35a with one inlet of the discrete phase Y-shaped mixing segment 32c, and the second discrete phase segment 32b connects the second discrete phase inlet 35b with the other inlet of the discrete phase Y-shaped mixing segment 32c; the outlet of the discrete phase Y-shaped mixing segment 32c is connected to the flow focusing structure 33, and the fluid in the first discrete phase segment 32a and the fluid in the second discrete phase segment 32b can be merged and flow into the flow focusing structure 33 through the discrete phase Y-shaped mixing segment 32c. In the above embodiment, the interface component 2 can have two discrete phase interfaces 2b, one of which is connected to all first discrete phase inlets 35a, and the other is connected to all second discrete phase inlets 35b.

[0098] Preferably, as shown in Figures 1 to 3 and 7, the two discrete phase interfaces 2b are both circular in shape, and a plurality of first discrete phase inlets 35a are arranged in parallel and evenly on the microfluidic chip 3, and all first discrete phase inlets 35a are in the shape of circular holes. The microfluidic chip 3 also has a plurality of second discrete phase inlets 35b arranged in parallel and evenly, and all second discrete phase inlets 35b are in the shape of circular holes. One discrete phase interface 2b is connected to all first discrete phase inlets 35a, and the other discrete phase interface 2b is connected to all second discrete phase inlets 35b. Those skilled in the art can flexibly set the number of first discrete phase inlets 35a and second discrete phase inlets 35b in actual applications, for example, 4 to 16. In the example of the invention, the number of first discrete phase inlets 35a and second discrete phase inlets 35b are both 8.

[0099] Preferably, referring to Figures 7 and 8, the downstream side of the flow focusing structure 33 has a gradually expanding section 37, and the width of the gradually expanding section 37 gradually increases along the flow direction of the liquid. Further preferably, the length of the outflow section 33d is 3d-6d, the arc radius of the gradually expanding section 37 is 4d, the length of the gradually expanding section 37 is 10d-15d, and the width of the downstream side of the gradually expanding section 37 is 4d-6d. With this arrangement, high-throughput microsphere preparation can be achieved, and the stability and uniformity of the microspheres can be improved. The microsphere generation rate is 1×10 8 -9×10 8 / hour, preferably, the microsphere generation rate is 2×10 8 / hour.

[0100] In some embodiments, referring to Figures 7 and 8, the continuous phase inlet 34 communicates with the inlet of the continuous phase Y-shaped diverter section 31a via a continuous connecting section 31d. The continuous connecting section 31d has a width of 3.2d. The first continuous phase diverter section 31a has a width of 1.6d at the beginning and a width of 1.6d at the end, which is inclined and gradually decreases from 1.6d to d. The second continuous phase diverter section 31b has a width of 1.6d at the beginning and a width of 1.6d at the end, which is inclined and gradually decreases from 1.6d to d. The continuous phase channel 31 may have a bend with a diameter of 10d.

[0101] Preferably, referring to Figures 7 and 8, the number of microsphere preparation channels is an even number, and all microsphere preparation channels are arranged in pairs, with the two microsphere preparation channels in each group being connected to the mixed phase outlet 36 through the same incorporation structure 38. Taking the example of eight microsphere preparation channels, microsphere preparation channel No. 1 and microsphere preparation channel No. 2 are incorporated into one channel; microsphere preparation channel No. 3 and microsphere preparation channel No. 4 are incorporated into one channel, and so on, microsphere preparation channel No. 7 and microsphere preparation channel No. 8 are incorporated into one channel, which facilitates ensuring that the downstream back pressure of each channel remains consistent. Further preferably, the two microsphere preparation channels in each group are arranged adjacent to each other. Such an arrangement facilitates the arrangement of all microsphere preparation channels and improves their compactness.

[0102] In one embodiment, each incorporation structure 38 communicates with the mixed-phase outlet 36 via an inflow channel 39. Each inflow channel 39 has at least one curved section, wherein the diameter of the curved section is preferably 10d, thereby ensuring consistent flow resistance in the inflow channel 39. Preferably, all inflow channels 39 have the same number of curved sections. For example, in the structure shown in FIG7 , each inflow channel 39 has six curved sections. In other embodiments, the number of curved sections can also be other numbers. In one example, the outlets of all inflow channels 39 are evenly arranged around the circumference of the mixed-phase outlet 36, thereby ensuring that the mixed phase flows out of the mixed-phase outlet 36 evenly.

[0103] Regarding the interface assembly 2, in one example, its dimensions can be designed based on material strength, generally ensuring that the thinnest portion is no less than 40d. The chip width tolerance is ±0.002d. The thickness tolerance between the interface assembly 2, the inlet gasket 4, the outlet gasket 5, and the microfluidic chip 3 should be maintained at +0.005d, ensuring sealing even with some gasket deformation. For the continuous phase inlet 34, the discrete phase inlet 35, and the mixed phase outlet 36, the corresponding gasket holes on the inlet gasket 4 and the outlet gasket 5 should be 8-12d larger than these phase inlets / outlets, ensuring maximum fluid flow even with alignment errors.

[0104] The present invention also provides a method for preparing microspheres based on the high-throughput microsphere preparation device 1, and the method for preparing microspheres comprises the following steps:

[0105] S1: dissolving a cross-linkable artificial synthetic biomaterial and / or a cross-linkable natural biomaterial in an aqueous solvent, and filtering the solution through a 0.22 μm filter membrane to form a continuous aqueous phase solution.

[0106] Among them, in some embodiments, the cross-linkable artificial synthetic biomaterial is selected from one or more of polyanhydrides, polyesters, polyamides, polyamino acids, polypropylene, polystyrene, polyacrylamide, polyethylene glycol, polyethylene glycol derivatives, polylactic acid, polyhydroxy acid, polylactic acid-alcohol copolymers, polypyrrole, polyester, polymethacrylate, polydimethylsiloxane, polyacetal, polycyanoacrylate, polyurethane, polyethylene, polycarbonate or polyethylene oxide, and the cross-linkable natural biomaterial is selected from one or more of gelatin, gelatin derivatives, alginate, alginate derivatives, collagen, proteoglycan, glycoprotein, agar, matrigel, hyaluronic acid, laminin, fibronectin, chitosan, chitosan derivatives, and soy protein.

[0107] S2: The organic oil phase reagent and the surfactant are fully mixed in a mass ratio of 3:1-5:1, and filtered through a 0.22 μm filter membrane to form a discrete phase oil phase solution.

[0108] In some embodiments, the organic oil phase reagent is selected from one or more of mineral oil, long-chain alkanes, and HFE7500 fluorinated oil, and the surfactant is Span 80 or Tween 80.

[0109] S3: Mix the crosslinking agent into the oil phase as the collection liquid.

[0110] In some embodiments, the cross-linking agent is selected from one or more of glutaraldehyde (GA), genipin, transglutaminase, tetramethylethylenediamine (TEMED), ammonium persulfate (APS), calcium sulfate, calcium chloride, zinc sulfate, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC).

[0111] S4: The continuous phase aqueous solution is pumped into the continuous phase interface 2a through the first pump and the aqueous phase flow rate is controlled to be 0.2-30 μL / min. The dispersed phase oil phase solution is pumped into the dispersed phase interface 2b through the second pump and the oil phase flow rate is controlled to be 20-1000 μL / min.

[0112] Preferably, the water phase flow rate is 7 μL / min, the oil phase flow rate is 40 μL / min, and the continuous water phase solution is the continuous water phase solution after air bubbles are removed, and the dispersed oil phase solution is the dispersed oil phase solution after air bubbles are removed.

[0113] S5: The formed microspheres are received in the collection liquid through the mixed phase interface 2c.

[0114] S6: Separate the microspheres by filtration, washing and centrifugation.

[0115] In a specific embodiment, which is not limiting, the premix solution in step S1 is configured as follows: gelatin is dissolved in deionized water to obtain a gelatin solution, wherein the gelatin content in the gelatin solution is 0.3 g of gelatin per 10 ml of the solution; the gelatin solution is filtered using a 0.22 μm filter membrane to form a continuous aqueous phase solution; the gelatin solution is drawn into a syringe, which is placed vertically to expel air bubbles and maintained for 15 minutes; the syringe is mounted on a syringe pump and connected to the interface component 2 via a Luer connector, and then connected to the microfluidic chip 3;

[0116] The oil phase premix solution in step S2 is configured as follows: the long-chain alkane organic oil phase and Span 80 are thoroughly mixed in a mass ratio of 3:1-5:1, filtered through a 0.22 μm filter membrane to form a discontinuous oil phase solution, the oil phase solvent is drawn into a syringe, and the syringe is placed vertically to expel air bubbles for 15 minutes. The syringe is then mounted on a syringe pump and connected to the interface component 2 via a Luer connector, and then connected to the microfluidic chip 3;

[0117] In step S3, GA is diluted at a ratio of 1:10 and mixed with the oil phase as the collection liquid;

[0118] After the oil phase and water phase syringes are installed on the syringe pump, the water phase flow rate is 7 μL / min and the oil phase flow rate is 40 μL / min. After the oil phase and water phase pass through the cross-intersection to form stable microspheres, the microspheres are collected and received in the oil phase containing the cross-linker;

[0119] In the above, referring to Figures 10 to 12, the diameter of the prepared single microspheres is 5-80 microns, and the flow rate of the water phase and the oil phase can be adjusted according to the target size.

[0120] Optionally, in the preparation of water-in-oil microspheres, the aqueous phase is the configured reaction liquid, and the oil phase can be mineral oil, long-chain alkanes, HFE7500 microfluidic special fluorinated oil, etc.

[0121] Optionally, in the preparation of oil-in-water microspheres, the aqueous phase is an alcohol hydrocarbon such as methanol, butanol, isopropanol, etc., and the oil phase can be mineral oil, olive oil, etc.

[0122] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A microfluidic chip, characterized in that: The microfluidic chip comprises a mixed phase outlet and a plurality of microsphere preparation channels which are connected to the mixed phase outlet and are arranged in parallel with each other; Each of the microsphere preparation channels comprises a continuous phase channel, a discrete phase channel and a flow focusing structure, wherein the continuous phase channel connects the continuous phase inlet with the flow focusing structure, the discrete phase channel connects the discrete phase inlet with the flow focusing structure, and the flow focusing structure connects the mixed phase outlet; The flow focusing structure is configured to disperse the dispersed phase fluid into a plurality of microspheres through the flow shear force of the main phase fluid in the continuous phase channel on the dispersed phase fluid in the discrete phase channel.

2. The microfluidic chip according to claim 1, characterized in that: The continuous phase channel includes a continuous phase Y-shaped flow dividing section, a first continuous phase flow dividing section, and a second continuous phase flow dividing section; The continuous phase inlet is connected to the inlet of the continuous phase Y-shaped flow dividing section, the two outlets of the continuous phase Y-shaped flow dividing section are connected to the first continuous phase flow dividing section and the second continuous phase flow dividing section respectively, and the first continuous phase flow dividing section and the second continuous phase flow dividing section are both connected to the flow focusing structure; The connecting position between the first continuous phase flow splitting section and the flow focusing structure and the connecting position between the second continuous phase flow splitting section and the flow focusing structure are respectively placed on both sides of the connecting position between the discrete phase channel and the flow focusing structure.

3. The microfluidic chip according to claim 2, characterized in that: The flow focusing structure is in a cross shape.

4. The microfluidic chip according to claim 2, characterized in that: The discrete phase channel includes a first discrete phase segment, a second discrete phase segment and a discrete phase Y-shaped mixing segment, and the discrete phase inlet includes a first discrete phase inlet and a second discrete phase inlet; The first discrete phase segment connects the first discrete phase inlet with one inlet of the discrete phase Y-shaped mixing segment, the second discrete phase segment connects the second discrete phase inlet with another inlet of the discrete phase Y-shaped mixing segment, and the outlet of the discrete phase Y-shaped mixing segment is connected to the flow focusing structure.

5. The microfluidic chip according to claim 2, characterized in that: The downstream side of the flow focusing structure has a gradually expanding section, and the width of the gradually expanding section gradually increases along the flow direction of the liquid.

6. The microfluidic chip according to claim 2, characterized in that: The number of the microsphere preparation channels is an even number, all the microsphere preparation channels are grouped in pairs, and the two microsphere preparation channels in each group are connected to the mixed phase outlet through the same incorporation structure.

7. The microfluidic chip according to claim 6, characterized in that: The two microsphere preparation channels in each group are arranged adjacent to each other.

8. The microfluidic chip according to claim 6, characterized in that: Each of the incorporation structures is communicated with the mixed phase outlet via an inflow channel, and each of the inflow channels has at least one bend section.

9. The microfluidic chip according to claim 8, characterized in that: All of the inflow channels have the same number of curved sections.

10. The microfluidic chip according to claim 8, characterized in that: The outlets of all the inflow channels are evenly arranged around the circumference of the mixed phase outlet.

11. The microfluidic chip according to claim 5, characterized in that: The flow focusing structure has a first continuous phase inflow section, a second continuous phase inflow section, a discrete phase inflow section and an outflow section, the first continuous phase inflow section is connected to the first continuous phase diversion section, the second continuous phase inflow section is connected to the second continuous phase diversion section, the discrete phase inflow section is connected to the discrete phase channel, and the first continuous phase inflow section, the second continuous phase inflow section and the discrete phase inflow section have the same width and the width is d.

12. The microfluidic chip according to claim 11, characterized in that: The length of the outflow section is 3d-6d; and / or, The arc radius of the gradually expanding section is 4d; and / or, The length of the gradually expanding section is 10d-15d; and / or, The width of the downstream side of the gradually expanding section is 4d-6d.

13. The microfluidic chip according to claim 11, characterized in that: The continuous phase inlet is connected to the inlet of the continuous phase Y-shaped diversion section through a continuously connected section, the width of the continuously connected section is 3.2d, the first section width of the first continuous phase diversion section is 1.6d, the tail section is inclined and the width gradually shrinks from 1.6d to d, the first section width of the second continuous phase diversion section is 1.6d, the tail section is inclined and the width gradually shrinks from 1.6d to d.

14. The microfluidic chip according to claim 11, characterized in that: The value range of d is 30-60 microns.

15. The microfluidic chip according to claim 14, characterized in that: d is 50 μm.

16. The microfluidic chip according to any one of claims 1 to 15, characterized in that: The microfluidic chip is made of stainless steel, COC, PMMA, PC, PE or glass.

17. A device for high-throughput preparation of microspheres, characterized in that: The device for preparing microspheres with high throughput comprises an interface component, a first pump component, a second pump component and a microfluidic chip according to any one of claims 1 to 16; The interface component has a continuous phase interface, a discrete phase interface and a mixed phase interface. The continuous phase interface is connected to the first pump component and the continuous phase inlet, the discrete phase interface is connected to the second pump component and the discrete phase inlet, and the mixed phase interface is connected to the mixed phase outlet.

18. The device for high-throughput preparation of microspheres according to claim 17, characterized in that: The interface component includes an inlet mechanism and an outlet mechanism, the continuous phase interface and the discrete phase interface are both arranged on the inlet mechanism, the mixed phase interface is arranged on the outlet mechanism, the part of the microfluidic chip having the continuous phase inlet and the discrete phase inlet is arranged on the inlet mechanism, and the part of the microfluidic chip having the mixed phase outlet is arranged on the outlet mechanism.

19. The device for high-throughput preparation of microspheres according to claim 18, characterized in that: The inlet mechanism comprises an inlet upper plate and an inlet lower plate connected to each other, the part of the microfluidic chip having the continuous phase inlet and the discrete phase inlet is sandwiched between the inlet upper plate and the inlet lower plate, and the continuous phase interface and the discrete phase interface are both arranged on the inlet upper plate.

20. The device for high-throughput preparation of microspheres according to claim 19, characterized in that: An inlet gasket is arranged between the inlet upper plate and the microfluidic chip, and a continuous phase buffer groove and a discrete phase buffer groove are formed on the inlet gasket. The continuous phase buffer groove connects the continuous phase interface with the continuous phase inlet, and the discrete phase buffer groove connects the discrete phase interface with the discrete phase inlet.

21. The device for high-throughput preparation of microspheres according to claim 20, characterized in that: A first gasket receiving groove is formed on the inlet upper plate, and a portion of the inlet gasket is received in the first gasket receiving groove.

22. The device for high-throughput preparation of microspheres according to claim 21, characterized in that: The first gasket receiving groove is provided with at least one positioning protrusion, and the inlet gasket is formed with positioning holes which correspond to the positioning protrusions one by one and are positioned and matched with each other.

23. The device for high-throughput preparation of microspheres according to claim 20, characterized in that: The inlet upper plate and the inlet lower plate are connected by a plurality of fastening bolts, so that the inlet gasket is sealed and fitted with the inlet upper plate and the microfluidic chip respectively.

24. The device for high-throughput preparation of microspheres according to claim 19, characterized in that: The inlet upper plate is provided with at least one positioning block, and the inlet lower plate is provided with a positioning groove which corresponds to and cooperates with the positioning block in a one-to-one manner.

25. The device for high-throughput preparation of microspheres according to claim 18, characterized in that: The outlet mechanism comprises an outlet upper plate and an outlet lower plate connected to each other, the part of the microfluidic chip having the mixed phase outlet is sandwiched between the outlet upper plate and the outlet lower plate, and the mixed phase interface is arranged on the outlet upper plate.

26. The device for high-throughput preparation of microspheres according to claim 25, characterized in that: An outlet gasket is arranged between the outlet upper plate and the microfluidic chip, and a mixed phase opening is formed on the outlet gasket, and the mixed phase opening connects the mixed phase interface with the mixed phase outlet.

27. The device for high-throughput preparation of microspheres according to claim 26, characterized in that: A second gasket accommodating groove is formed on the outlet upper plate, and a portion of the outlet gasket is accommodated in the second gasket accommodating groove.

28. The device for high-throughput preparation of microspheres according to claim 26, characterized in that: The outlet upper plate and the outlet lower plate are connected by a plurality of fastening bolts, so that the outlet gasket is sealed and fitted with the outlet upper plate and the microfluidic chip respectively.

29. The device for high-throughput preparation of microspheres according to any one of claims 17 to 28, characterized in that: The shapes of the continuous phase inlet and the discrete phase inlet are both circular holes, all the continuous phase inlets are evenly arranged in parallel, and all the discrete phase inlets are evenly arranged in parallel.

30. A method for preparing microspheres based on the high-throughput microsphere preparation device according to any one of claims 17 to 29, characterized in that: The method for preparing microspheres comprises the following steps: S1: dissolving the cross-linkable artificial synthetic biomaterial and / or the cross-linkable natural biomaterial in an aqueous phase solvent, and filtering with a 0.22 μm filter membrane to form a continuous phase aqueous phase solution; S2: the organic oil phase reagent and the surfactant are fully mixed in a mass ratio of 3:1-5:1, and filtered through a 0.22 μm filter membrane to form a discrete phase oil phase solution; S3: mixing the crosslinker into the oil phase as the collection liquid; S4: pumping the continuous phase aqueous solution into the continuous phase interface through the first pump and controlling the aqueous phase flow rate to be 0.2-30 μL / min, and pumping the dispersed phase oil phase solution into the dispersed phase interface through the second pump and controlling the oil phase flow rate to be 20-1000 μL / min; S5: receiving the formed microspheres into the collection liquid through the mixed phase interface; S6: Separate the microspheres by filtration, washing and centrifugation.

31. The method for preparing microspheres according to claim 30, characterized in that: The cross-linkable artificial synthetic biomaterial is selected from one or more of polyanhydrides, polyesters, polyamides, polyamino acids, polypropylene, polystyrene, polyacrylamide, polyethylene glycol, polyethylene glycol derivatives, polylactic acid, polyhydroxy acid, polylactic acid-alkyd copolymers, polypyrrole, polyester, polymethacrylate, polydimethylsiloxane, polyacetal, polycyanoacrylate, polyurethane, polyethylene, polycarbonate or polyethylene oxide.

32. The method for preparing microspheres according to claim 30, characterized in that: The cross-linkable natural biomaterial is selected from one or more of gelatin, gelatin derivatives, alginate, alginate derivatives, collagen, proteoglycan, glycoprotein, agar, matrigel, hyaluronic acid, laminin, fibronectin, chitosan, chitosan derivatives, and soy protein.

33. The method for preparing microspheres according to claim 30, characterized in that: The organic oil phase reagent is selected from one or more of mineral oil, long-chain alkanes, and HFE7500 fluorinated oil.

34. The method for preparing microspheres according to claim 30, characterized in that: The surfactant is Span 80 or Tween 80.

35. The method for preparing microspheres according to claim 30, characterized in that: The cross-linking agent is selected from one or more of glutaraldehyde, genipin, transglutaminase, tetramethylethylenediamine, ammonium persulfate, calcium sulfate, calcium chloride, zinc sulfate, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.

36. The method for preparing microspheres according to claim 30, characterized in that: The aqueous phase flow rate is 7 μL / min; and / or, The oil phase flow rate was 40 μL / min.

37. The method for preparing microspheres according to any one of claims 30 to 36, characterized in that: In step S4, the continuous phase aqueous solution is a continuous phase aqueous solution after air bubbles are exhausted, and the dispersed phase oil phase solution is a dispersed phase oil phase solution after air bubbles are exhausted.

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