Micro-fluidic chip, device for preparing microspheres in high flux and method for preparing microspheres

Abstract

The invention provides a micro-fluidic chip, a high-flux microsphere preparation device and a microsphere preparation method. The micro-fluidic chip comprises a mixed phase outlet and a plurality of microsphere preparation channels which are communicated with the mixed phase outlet and are arranged in parallel, each microsphere preparation channel comprises a continuous phase channel, a dispersed phase channel and a flow focusing structure, the continuous phase inlet is communicated with the flow focusing structure through the continuous phase channel, the dispersed phase inlet is communicated with the flow focusing structure through the dispersed phase channel, and the flow focusing structure is communicated with the mixed phase outlet; the flow focusing structure is arranged to be capable of dispersing the dispersed-phase fluid into a plurality of microspheres under the action of flow shearing force of the main-phase fluid in the continuous-phase channel on the dispersed-phase fluid in the dispersed-phase channel. The high-throughput microsphere preparation device comprises an interface assembly, a first pump part, a second pump part and a micro-fluidic chip. According to the invention, high-throughput preparation performance can be provided, and the requirements on large-order-of-magnitude microspheres in the fields of biology, chemistry and the like can be met.

Description

Technical Field

[0001] The 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

[0002] Microfluidics is an emerging discipline, which mainly benefits from the progress of micro-nano level technology and the urgent need for new materials in the biochemical field. It designs and manipulates chemical and biological processes at the microscale, and shows great potential in dealing with obstacles that traditional chemical and biological methods cannot handle. In the early days, microfluidics was also called micro-total analysis laboratory, which could integrate some or even all functions of the laboratory into a chip for separation, mixing, and detection of samples; with the progress of design process, micro-nano processing, labeling and other technologies, microfluidics is no longer limited to micro-total analysis, but extends its tentacles to many aspects of biochemistry, physiology and other disciplines. Especially after more than 20 years of rapid development, microfluidics has played an important role in the fields of biology, chemical engineering, and pharmaceuticals. It can not only serve as a substitute for existing technologies, such as cell culture, emulsification, and drug encapsulation, but also as a new technology to provide new industrial efficiency improvement points, such as microreactors, fine chemicals, POCT chips, etc.

[0003] A typical application of microfluidics is to prepare microspheres. By using a continuous phase shear fluid to overcome the surface tension and discretize it, small microspheres are formed. In the presence of a surfactant, the microspheres can exist stably. Microspheres have a variety of applications. They can be used as microreactors to control various biochemical reactions in microscale microspheres; they can be used to encapsulate cells to provide the necessities for cells to maintain activity; they can also be used for targeted drug delivery, modifying chemokines on the surface of microspheres to deliver drugs to the target area in a directionally controlled manner. 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 shearing effect of a continuous phase on another phase to form discrete microspheres.

[0004] Microspheres can be either water-in-oil or oil-in-water, with continuous and discrete phases set according to specific usage scenarios. Oil-in-water microspheres are often involved, and the aqueous solution in the water microspheres can be either an aqueous solvent solution or a hydrogel solution. In the biomedical field, hydrogel solutions are the most common. Hydrogel is a material with good biocompatibility, more than 80% of its volume is free water, so it can provide a living environment for cells and other biological elements; and because hydrogels can have different cross-linking methods and degrees of cross-linking, they can provide a wide range of mechanical properties and are used to prepare various biomaterials, such as drug capsules, microtissues, culture carriers, etc. Commonly used hydrogels include PEGS hydrogels, PLAG hydrogels, dextran, and alginate hydrogels. Different hydrogel systems can be designed according to different application scenarios based on whether they are degradable, temperature sensitive, and whether they can respond to pH.

[0005] Since microfluidic chips can provide size-controlled aqueous microspheres at high throughput, they can be widely used in the preparation of hydrogel microspheres. However, most of the current preparation methods are single-channel, with frequencies ranging from hundreds of thousands of hertz, while the microspheres used in the biological field require orders of magnitude 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, 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. Summary of the invention

[0006] In view of the shortcomings of the prior art, the problem of low throughput in microsphere preparation and too high lower limit of microsphere diameter adjustment is solved, so as to meet the high-throughput preparation of microspheres and provide a wider adjustment range of microsphere diameter.

[0007] According to a first aspect of the present invention, there is provided a microfluidic chip, the 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;

[0008] 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;

[0009] 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.

[0010] Preferably, the continuous phase channel comprises a continuous phase Y-shaped flow dividing section, a first continuous phase flow dividing section and a second continuous phase flow dividing section;

[0011] 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;

[0012] 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.

[0013] Preferably, the flow focusing structure is in a cross shape.

[0014] 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;

[0015] 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.

[0016] 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.

[0017] 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.

[0018] Preferably, the two microsphere preparation channels in each group are arranged adjacent to each other.

[0019] Preferably, each of the incorporation structures is connected to the mixed phase outlet via an inflow channel, and each of the inflow channels has at least one bend section.

[0020] Preferably, all the inflow channels have the same number of curved sections.

[0021] Preferably, the outlets of all the inflow channels are evenly arranged around the circumference of the mixed phase outlet.

[0022] 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.

[0023] Preferably, the length of the outflow section is 3d-6d; and / or,

[0024] The arc radius of the gradually expanding section is 4d; and / or,

[0025] The length of the gradually expanding section is 10d-15d; and / or,

[0026] The width of the downstream side of the gradually expanding section is 4d-6d.

[0027] 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 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.

[0028] Preferably, the value range of d is 30-60 microns.

[0029] Preferably, d is 50 microns.

[0030] Preferably, the microfluidic chip is made of stainless steel, COC, PMMA, PC, PE or glass.

[0031] 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 component, a first pump component, a second pump component and the microfluidic chip described in any one of the above;

[0032] 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.

[0033] 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.

[0034] Preferably, the inlet mechanism comprises 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 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.

[0035] Preferably, 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.

[0036] Preferably, a first gasket accommodating groove is formed on the inlet upper plate, and a portion of the inlet gasket is accommodated in the first gasket accommodating groove.

[0037] 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 protrusions and are positioned and matched with each other.

[0038] 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.

[0039] Preferably, at least one positioning block is provided on the inlet upper plate, and a positioning groove corresponding to and cooperating with the positioning block is provided on the inlet lower plate.

[0040] Preferably, the outlet mechanism comprises an outlet upper plate and an outlet lower plate connected to each other, 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.

[0041] Preferably, an outlet gasket is provided between the outlet upper plate and the microfluidic chip, 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.

[0042] Preferably, 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.

[0043] 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.

[0044] 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.

[0045] According to a third aspect of the present invention, there is provided a method for preparing microspheres based on the high-throughput microsphere preparation device described in any one of the above, the method for preparing microspheres comprising the following steps:

[0046] 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;

[0047] 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;

[0048] S3: mixing the crosslinker into the oil phase as the collection liquid;

[0049] 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;

[0050] S5: receiving the formed microspheres into the collection liquid through the mixed phase interface;

[0051] S6: Separate the microspheres by filtration, washing and centrifugation.

[0052] 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-alkyd copolymers, polypyrrole, polyester, polymethacrylate, polydimethylsiloxane, polyacetal, polycyanoacrylate, polyurethane, polyethylene, polycarbonate or polyethylene oxide.

[0053] 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.

[0054] Preferably, the organic oil phase reagent is selected from one or more of mineral oil, long-chain alkanes, and HFE7500 fluorinated oil.

[0055] Preferably, the surfactant is Span 80 or Tween 80.

[0056] 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.

[0057] Preferably, the aqueous phase flow rate is 7 μL / min; and / or,

[0058] The oil phase flow rate was 40 μL / min.

[0059] 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.

[0060] 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 and provide 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 needs of biology, chemistry and other fields for large-scale microspheres. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the following description of the embodiments in conjunction with the accompanying drawings, in which:

[0062] Figure 1 It is a schematic diagram of the overall structure of the device for preparing microspheres with high throughput of the present invention;

[0063] Figure 2 is an exploded view of a device for preparing microspheres with high throughput according to the present invention;

[0064] Figure 3 The structure of the inlet upper plate in the inlet mechanism of the interface assembly of the high-throughput microsphere preparation device of the present invention is schematically shown. Figure 1 ;

[0065] Figure 4 The structure of the inlet upper plate in the inlet mechanism of the interface assembly of the high-throughput microsphere preparation device of the present invention is schematically shown. Figure 2 ;

[0066] Figure 5 The structure of the outlet upper plate in the outlet mechanism of the interface assembly of the high-throughput microsphere preparation device of the present invention is schematically shown. Figure 1 ;

[0067] Figure 6 The structure of the outlet upper plate in the outlet mechanism of the interface assembly of the high-throughput microsphere preparation device of the present invention is schematically shown. Figure 2 ;

[0068] Figure 7 It is a schematic diagram of the overall structure of the microfluidic chip of the present invention;

[0069] Figure 8 is a partial schematic diagram of the microfluidic chip of the present invention;

[0070] Fig. 9 It is a schematic structural diagram of an inlet gasket of a high-throughput microsphere preparation device of the present invention;

[0071] Fig.10 It is a light microscopic image of a plurality of microspheres prepared by the high-throughput microsphere preparation device of the present invention;

[0072] Fig.11 It is an electron microscope image of a plurality of microspheres prepared by the high-throughput microsphere preparation device of the present invention;

[0073] Fig.12 This is an electron microscope image of a single microsphere prepared by the high-throughput microsphere preparation device of the present invention.

[0074] List of reference numerals:

[0075] 1. Device for high-throughput preparation of microspheres;

[0076] 2. Interface components; 2a. Continuous phase interface; 2b. Discrete phase interface; 2c. Mixed phase interface;

[0077] 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;

[0078] 22, outlet mechanism; 221, outlet upper plate; 221a, second gasket receiving groove; 222, outlet lower plate;

[0079] 3. Microfluidic chip;

[0080] 31, continuous phase channel; 31a, continuous phase Y-shaped flow dividing section; 31b, first continuous phase flow dividing section; 31c, second continuous phase flow dividing section; 31d, continuous phase connecting section;

[0081] 32, discrete phase channel; 32a, first discrete phase segment; 32b, second discrete phase segment; 32c, discrete phase Y-shaped mixing segment;

[0082] 33, flow focusing structure; 33a, first continuous phase inflow section; 33b, second continuous phase inflow section; 33c, discrete phase inflow section; 33d, outflow section;

[0083] 34. continuous phase inlet; 35. discrete phase inlet; 35a. first discrete phase inlet; 35b. second discrete phase inlet; 36. mixed phase outlet; 37. gradually expanding section; 38. merged structure; 39. inflow channel;

[0084] 4. Inlet gasket; 4a. Continuous phase buffer groove; 4b. Discrete phase buffer groove; 4c. Positioning hole;

[0085] 5. Export gasket. DETAILED DESCRIPTION

[0086] 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.

[0087] 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 cannot be regarded as limiting the scope of protection of this application.

[0088] In the description of the present invention, it is to 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”, etc., indicating orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as limiting the present invention.

[0089] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the present invention, the meaning of "plurality" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined.

[0090] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral one; it can be a mechanical connection, an electrical connection, or communication with each other; it can be a direct connection, or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0091] In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may mean that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, a first feature being "above", "above" or "above" a second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. A first feature being "below", "below" or "below" a second feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.

[0092] In the present invention, the terms "one embodiment", "some embodiments", "examples", "specific examples", or "some examples" etc. 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 representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the described specific features, structures, materials or characteristics may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0093] like Figures 1 to 7As shown, 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 connected to the mixed phase outlet 36 and arranged in parallel with each other, and the flow and pressure of each microsphere preparation channel are ensured to be consistent by controlling the flow resistance, 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 the continuous phase inlet 34, the discrete phase interface 2b is connected to the second pump component and 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 requirements can also be selected. The advantage of such a configuration is that each interface corresponds to a power source, saving costs.

[0094] In some embodiments, see Figures 1 to 3 5, the continuous phase interface 2a, the discrete phase interface 2b and the mixed phase interface 2c all have a sinking mechanism, which can be formed on the interface boss, and the sinking structure is formed with a threaded interface, which can be connected to a threaded pipe, and the threaded pipe is then connected to the corresponding microinjection pump. Exemplarily, the microinjection pump is connected to the threaded interface through the threaded pipe, so as to achieve the connection between the microinjection pump and the continuous phase interface 2a. In practical applications, the connection between the pipeline and the interface can be achieved through a Luer connector to ensure sealing. The threaded interface can be selected in a specification of 1 / 4-28UN F. In other examples, the interface can also be connected to the corresponding pipeline by a snap-on method.

[0095] In one embodiment, see Figure 1 and 2The 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 arranged on the inlet mechanism 21, the mixed phase interface 2c is arranged on the outlet mechanism 22, the part of the microfluidic chip 3 having the continuous phase inlet 34 and the discrete phase inlet 35 is arranged on the inlet mechanism 21, and the part of the microfluidic chip 3 having the mixed phase outlet 36 is arranged on the outlet mechanism 22. In a specific example, the inlet mechanism 21 and the outlet mechanism 22 adopt a split design, exposing the flow focusing structure 33 of the microfluidic chip 3 between the inlet mechanism 21 and the outlet mechanism 22, reducing the difficulty of assembly, saving materials, and reducing material costs.

[0096] Preferably, see Figures 1 to 4 , 7 and 9, the inlet mechanism 21 includes an inlet upper plate 211 and an inlet lower plate 212 connected to each other, the part 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, and the continuous phase interface 2a and the discrete phase interface 2b are both arranged on the inlet upper plate 211. Further preferably, an inlet gasket 4 is arranged between the inlet upper plate 211 and the microfluidic chip 3, and a continuous phase buffer groove 4a and a discrete phase buffer groove 4b are formed on the inlet gasket 4, 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.

[0097] 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 a plurality of fastening bolts ( Figures 1 to 3 As shown in the figure, the inlet gasket 4 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 easy to disassemble and clean. 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, and the inlet gasket 4 can be limited to a certain extent by the first gasket receiving groove 211a. Further preferably, at least one positioning protrusion 211b is provided in the first gasket receiving groove 211a, and a positioning hole 4c corresponding to the positioning protrusion 211b and cooperating with each other is formed on the inlet gasket 4, Figure 2 and 4 As shown in FIG. , two positioning protrusions 211 b are positioned and matched with two positioning holes 4 c in a one-to-one correspondence.

[0098] 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 cooperating with the positioning block 211c is provided on the inlet lower plate 212. Figure 2 and 4 As shown in FIG. , two positioning blocks 211 c and two positioning grooves 212 c are positioned and matched in a one-to-one correspondence.

[0099] Preferably, see Figure 1 , 2 , 5 to 7, the outlet mechanism 22 includes an outlet upper plate 221 and an outlet lower plate 222 connected to each other, the portion of the microfluidic chip 3 having the mixed phase outlet 36 is sandwiched between the outlet upper plate 221 and the outlet lower plate 222, and the mixed phase interface 2c is arranged on the outlet upper plate 221. Further preferably, an outlet gasket 5 is arranged between the outlet upper plate 221 and the microfluidic chip 3, and a mixed phase through hole is formed on the outlet gasket 5, and the mixed phase through hole connects the mixed phase interface 2c with the mixed phase outlet 36, and the mixed phase through hole can ensure that the liquid flowing into the mixed phase outlet 36 flows to the mixed phase interface 2c.

[0100] 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 a plurality of fastening bolts ( Figure 1 , 2 5 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, and 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.

[0101] In the above embodiment, the material of the inlet gasket 4 and the outlet gasket 5 can be perfluororubber (FFKM) or fluororubber (FKM), which has good corrosion resistance and can provide durable sealing. The microfluidic chip 3 can be made of stainless steel, COC, PMMA, PC, PE, glass and other materials, and the processing method can be photolithography, injection molding, laser processing and the like.

[0102] Preferably, Figures 1 to 3As shown in Figures 7 and 8, the continuous phase interface 2a is circular in shape, and a plurality of continuous phase inlets 34 are evenly and parallelly arranged 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 hole-shaped. Those skilled in the art can flexibly set the number of the continuous phase inlets 34 in practical applications, for example, 4 to 16. In the example of the invention, the number of the continuous phase inlets 34 is 8.

[0103] The following further describes the preferred embodiment of the microfluidic chip 3 of the present invention. Figure 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 connecting position of the first continuous phase diversion section 31b and the flow focusing structure 33 and the connecting position of the second continuous phase diversion section 31c and the flow focusing structure 33 are respectively placed on both sides of the connecting 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, and 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 through the continuous phase Y-shaped diversion section 31a, 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.

[0104] Preferably, the flow focusing structure 33 is in a cross shape. Figure 8 The flow focusing structure 33 has 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 where the first continuous phase inflow section 33a and the second continuous phase inflow section 33b are located is perpendicular to the straight line where the discrete phase inflow section 33c and the outflow section 33d are located, 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, 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 and are all 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.

[0105] Preferably, see Figure 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 another inlet of the discrete phase Y-shaped mixing segment 32c, and 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 the first discrete phase inlets 35a, and the other discrete phase interface 2b is connected to all the second discrete phase inlets 35b.

[0106] Preferably, Figures 1 to 3 As shown in , 7, the shapes of the two discrete phase interfaces 2b are both circular, and a plurality of first discrete phase inlets 35a are arranged in parallel and evenly on the microfluidic chip 3, and all the 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 the second discrete phase inlets 35b are in the shape of circular holes. One discrete phase interface 2b is connected to all the first discrete phase inlets 35a, and the other discrete phase interface 2b is connected to all the second discrete phase inlets 35b. Those skilled in the art can flexibly set the number of the first discrete phase inlets 35a and the second discrete phase inlets 35b in practical applications, for example, 4 to 16. In the example of the invention, the number of the first discrete phase inlets 35a and the second discrete phase inlets 35b are both 8.

[0107] Preferably, see Figure 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. Through such a setting, high-throughput preparation of microspheres 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.

[0108] In some embodiments, see Figure 7 and8 The continuous phase inlet 34 is connected to the inlet of the continuous phase Y-shaped flow dividing section 31a through the continuous connecting section 31d, the width of the continuous connecting section 31d is 3.2d, the first section width of the first continuous phase flow dividing section 31a is 1.6d, the tail section is inclined and the width gradually shrinks from 1.6d to d, and the first section width of the second continuous phase flow dividing section 31b is 1.6d, the tail section is inclined and the width gradually shrinks from 1.6d to d. Among them, the continuous phase channel 31 can have a bend, and the diameter of the bend is 10d.

[0109] Preferably, see Figure 7 and 8 , the number of microsphere preparation channels is an even number, all microsphere preparation channels are arranged in pairs, and the two microsphere preparation channels in each group are connected to the mixed phase outlet 36 through the same incorporation structure 38. Taking the number of microsphere preparation channels as an example, 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 is convenient for 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. Through such an arrangement, it is convenient to arrange all microsphere preparation channels and improve their compactness.

[0110] In one embodiment, each incorporation structure 38 is connected to the mixed phase outlet 36 via an inflow channel 39, and each inflow channel 39 has at least one bend section, wherein the diameter of the bend section is preferably 10d, thereby ensuring that the flow resistance in the inflow channel 39 is consistent. Preferably, the number of bend sections of all inflow channels 39 is the same, for example Figure 7 In the structure shown in , the number of the curved sections of the inflow channel 39 is 6. In other embodiments, the number of the curved sections can also be other numbers. In one example, the outlets of all the inflow channels 39 are evenly arranged around the circumference of the mixed phase outlet 36, so as to ensure that the mixed phase flows out of the mixed phase outlet 36 evenly.

[0111] For the interface component 2, in one example, the size of the interface component 2 can be designed according to the material strength, generally to ensure that the thinnest part is not less than 40d, the tolerance of the chip width matching is ±0.002d, and the thickness matching tolerance of the interface component 2 and the inlet gasket 4, the outlet gasket 5 and the microfluidic chip 3 should be guaranteed to be +0.005d, so that the sealing can be guaranteed even if the gasket undergoes a certain 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 larger than these phase inlets / outlets by 8-12d, so as to ensure that even if there is a misalignment, the maximum fluid flux can still flow.

[0112] 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:

[0113] 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.

[0114] 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, matrix glue, hyaluronic acid, laminin, fibronectin, chitosan, chitosan derivatives, and soy protein.

[0115] S2: The organic oil phase reagent and the surfactant are fully mixed in a mass ratio of 3:1-5:1, and filtered using a 0.22 μm filter membrane to form a discrete phase oil phase solution.

[0116] 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.

[0117] S3: Mix the crosslinking agent in the oil phase as the collection liquid.

[0118] 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).

[0119] S4: Pump the continuous phase aqueous solution into the continuous phase interface 2a through the first pump and control the aqueous phase flow rate to 0.2-30 μL / min, and pump the dispersed phase oil phase solution into the dispersed phase interface 2b through the second pump and control the oil phase flow rate to 20-1000 μL / min.

[0120] Preferably, the water phase flow rate is 7 μL / min, and the oil phase flow rate is 40 μL / min. In addition, the continuous phase water phase solution is the continuous phase water phase solution after the air bubbles are exhausted, and the dispersed phase oil phase solution is the dispersed phase oil phase solution after the air bubbles are exhausted.

[0121] S5: receiving the formed microspheres into the collection liquid through the mixed phase interface 2c.

[0122] S6: Separate the microspheres by filtration, washing and centrifugation.

[0123] In a specific embodiment, which is not limiting, the premixed 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 solution, the gelatin solution is filtered using a 0.22 μm filter membrane to form a continuous phase aqueous phase solution, the gelatin solution is sucked into a syringe, and the syringe is placed vertically to exhaust air bubbles for 15 minutes, the syringe is installed on a syringe pump, and connected to the interface component 2 through a Luer connector, and connected to the microfluidic chip 3;

[0124] The oil phase premix solution in step S2 is configured as follows: the long-chain alkane organic oil phase and Span 80 are fully mixed in a mass ratio of 3:1-5:1, filtered using a 0.22 μm filter membrane to form a discontinuous oil phase solution, the oil phase solvent is sucked into a syringe, and the syringe is placed vertically to exhaust air bubbles for 15 minutes, and the syringe is installed on a syringe pump and connected to the interface component 2 through a Luer connector, and connected to the microfluidic chip 3;

[0125] In step S3, GA is diluted at a ratio of 1:10 and mixed in the oil phase as a collection liquid;

[0126] 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 the water phase pass through the cross intersection to stably form microspheres, the microspheres are collected and received in the oil phase containing the cross-linking agent;

[0127] In the above, see Figures 10 to 12 The diameter of the prepared single microsphere is 5-80 microns, and the flow rate of the water phase and the oil phase can be adjusted according to the target size.

[0128] Optionally, in the preparation of water-in-oil microspheres, the water phase is the configured reaction liquid, and the oil phase can be mineral oil, long-chain alkanes, HFE7500 microfluidic special fluorinated oil, etc.

[0129] Optionally, in the preparation of oil-in-water microspheres, the water phase is an alcohol hydrocarbon such as methanol, butanol, isopropanol, etc., and the oil phase can be mineral oil, olive oil, etc.

[0130] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary 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 high-throughput microsphere preparation device 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 positioning grooves corresponding to and cooperating with the positioning blocks one by one.

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 solvent, and filtering with a 0.22 μm filter membrane to form a continuous 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 in 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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