High-flux micro-nano fluid control device and manufacturing method and application thereof

Abstract

The invention provides a high-throughput micro-nano fluid control device as well as a manufacturing method and application thereof. The high-throughput micro-nano fluid control device specifically comprises a sample channel, a micro-channel array and a compression channel array, the sample channel is a channel which is communicated with an inlet and a flow control outlet of the sample channel and is used for conveying a sample; the micro-channel array comprises at least one micro-channel and a port connected with the micro-channel; the compression channel array comprises at least one compression channel; the micro-channel is connected with the sample channel through the compression channel; the flow control outlet can be opened or closed in a controlled mode. The invention further provides a system comprising the control device and a preparation method. The control device can be used for loading exogenous therapeutic substances in the biological particles, preparing lipidosome, mixing samples and sorting the biological particles.

Description

Technical Field

[0001] The present invention relates to the field of microfluidics, and specifically provides a high-throughput microfluidic control device, a manufacturing method thereof, and an application thereof. Background Art

[0002] Drug delivery carriers are the basic methods to achieve precise drug transmission and targeted therapy in the body. Their development not only provides new research tools for precision medicine but also offers more efficient and safer means for disease treatment. Advanced drug delivery carriers aim to safely and efficiently target exogenous drugs to the lesion site, thereby reducing the toxic and side effects caused by the systemic distribution of traditional drugs. It is an important technology for personalized treatment and has important academic significance and practical value for the development of precision medicine and precision treatment.

[0003] As an important drug delivery carrier, lipid nanoparticles have greatly promoted the rapid clinical application of nucleic acid drugs and the wide use of mRNA vaccines, and are a representative case of advanced delivery systems promoting the development of the biomedical industry. In addition to the above chemically synthesized nanocarriers, extracellular vesicles (or exosomes), which are secreted by cells into the extracellular microenvironment and mediate intercellular material transport, have become a more promising drug delivery carrier due to their good biocompatibility, higher in vivo circulation stability, and powerful biological functions, and have received extensive attention from the international scientific research and industrial communities.

[0004] Although biological particles such as extracellular vesicles show great application prospects in the fields of intelligent drug delivery and precise disease diagnosis and treatment, they are still challenged and restricted by the key technology of efficient drug loading in biomedical research and clinical practice. There is an urgent need to develop a class of platform technologies that can load various exogenous substances and therapeutic drugs onto biological particles such as extracellular vesicles to ensure that the drug loading reaches the therapeutic effect, and the preparation process is stable, precise, and controllable. If the above technical bottlenecks are not broken through, even with advanced drug development plans, it is difficult to meet the major requirements for the stable production and control of drug delivery carriers, thus greatly limiting the clinical application prospects of drugs.

[0005] Microfluidic technology is a technology for studying and applying fluids at the micron or nanometer scale, which can precisely operate and control fluids and their contained substances through micro-nano scale channels or structures. Due to its characteristic size being similar to that of subcellular structures, microfluidic technology can precisely locate an external physical field to the nanoscale and exert more precise and uniform physical effects on biological particles such as extracellular vesicles, showing great application potential in the fields of separation, detection, processing, artificial synthesis, and exogenous substance loading of biological particles such as extracellular vesicles.

[0006] In the previous technical solutions, Chinese Patent Publication No.: CN110975953A discloses a microfluidic chip, its preparation method and application. This solution uses a compression channel structure with a height similar to the size of biological particles such as extracellular vesicles to apply a mechanical squeezing effect on biological particles such as extracellular vesicles, thereby achieving the loading of exogenous substances. In addition, Chinese Patent Publication No.: CN117920367A discloses a high-throughput micro-nano device based on the synergistic effect of mechanical and electrical forces, its preparation method and application. This solution uses the synergistic effect of electric field and mechanical squeezing to achieve membrane perforation of biological particles such as extracellular vesicles and load exogenous substances of different sizes.

[0007] However, there are still problems such as unreasonable structural layout in the preparation and application of drug delivery carriers in the above two technical solutions. Due to design defects such as the size difference between the microchannels and compression channels, the lack of mixed micro-nano structures inside the channels, and the sealing of the end of the inlet microchannel, the device still faces many technical problems such as bubble problems, channel blockage, sample dead volume, and device damage when processing biological particle samples such as extracellular vesicles.

[0008] The above-mentioned prior art has the following technical defects:

[0009] (1) There is a huge size difference between the microscale microchannels and nanoscale compression channels in the prior art solutions. When biological particles such as extracellular vesicles and exogenous substances inside the microchannels flow with the fluid, only a small part of the sample is transported to the smaller-sized compression channels, and a large amount of the sample remains inside the microchannels. This non-uniform sample distribution ultimately leads to sample accumulation at the end of the microchannel, blocking the compression channels near this position, and then causing the occlusion of the compression channels and a sharp increase in the internal fluid pressure of the chip, resulting in damage to the micro-nano device.

[0010] (2) In the prior art solutions, the end of the inlet microchannel adopts a closed design. After the sample solution enters the inlet microchannel, a large number of bubbles are formed by the gas inside the channel. Due to the closed structure at the end of the inlet microchannel, these bubbles cannot be effectively discharged, seriously blocking the junction of the compression channel and the inlet microchannel, resulting in the sample solution being unable to smoothly enter the compression channel, further exacerbating the sample accumulation at the end of the microchannel and the blockage of the compression channel, and ultimately leading to the chip breaking and failing.

[0011] (3) When the sample solution in the prior art solutions flows in the inlet microchannel, biological particles such as extracellular vesicles and exogenous substances mainly move to the end of the microchannel in a translational manner, rather than being evenly distributed and entering the compression channel, thus affecting the uniformity and stability of the preparation of drug delivery carriers.

[0012] (4) In the design of the device structure of the prior art solution, there is a gap distance between the end of the inlet microchannel and the compression channel, resulting in difficulty for the sample substance at the end of the inlet microchannel to enter the compression channel in real time. The samples accumulated for a long time are prone to deterioration / denaturation, thus contaminating the prepared drug delivery carrier and affecting its quality and stability. Summary of the Invention

[0013] To solve the problems existing in the prior art, the present invention provides a high-throughput micro-nano fluidic control device with a microchannel, a compression channel, and a sample channel. It can solve the problems existing in the prior art, precisely manipulate and process biological particles such as extracellular vesicles at the nanoscale with high throughput, achieve reversible perforation of biological particles such as extracellular vesicles and loading of exogenous substances, thereby efficiently, uniformly, and stably preparing a drug delivery carrier, and solving technical problems such as bubble problems, channel blockage, sample dead volume, and contamination of the delivery carrier existing in the existing micro-nano chips.

[0014] The first aspect of the present invention provides a high-throughput micro-nano fluidic control device, and the high-throughput micro-nano fluidic control device includes a sample channel, a microchannel array, and a compression channel array;

[0015] The sample channel is a channel that connects the inlet of the sample channel and the flow control outlet and is used to transport the sample;

[0016] The microchannel array includes at least one microchannel and a port connected to the microchannel;

[0017] The compression channel array includes at least one compression channel;

[0018] The microchannel is connected to the sample channel through the compression channel;

[0019] The flow control outlet can be controllably opened or closed.

[0020] Furthermore, the high-throughput micro-nano fluidic control device includes at least one group of microchannel arrays, preferably two or more groups.

[0021] Furthermore, the high-throughput micro-nano fluidic control device includes two groups of microchannel arrays and is arranged in an interdigitated and staggered manner.

[0022] Furthermore, the microchannel array includes two or more microchannels.

[0023] Furthermore, the flow control outlet is one or more.

[0024] Furthermore, it includes at least one sample channel.

[0025] Furthermore, the sample channel is a channel that includes one or more inlets connecting the sample channel and one or more flow control outlets.

[0026] Further, the channel of the sample channel is a straight line, a curve, a broken line, a fork or a radial shape.

[0027] Further, the microchannels are arranged around the sample channel, and preferably, the compression channel lengths between all the microchannels and the sample channel are equal or unequal.

[0028] Further, the difference in the lengths of different compression channels is within 50%.

[0029] Further, each of the microchannels is connected to the sample channel through at least one compression channel.

[0030] Further, the high-throughput micro-nano fluid control device is at least one layer, preferably one layer or more than two layers.

[0031] Further, when multiple layers of the high-throughput micro-nano fluid control device are provided, each layer can be provided with a sample channel, a microchannel array and a compression channel separately.

[0032] Further, micro-nano structures are provided in the sample channel.

[0033] Further, micro-nano structures are provided in the microchannels.

[0034] Further, nano-structures are provided in the compression channels.

[0035] Further, the inlet and the flow control outlet of the sample channel are connected through a circulation module, and the circulation module is used to circulate the sample flowing out of the flow control outlet back into the sample channel through the inlet of the sample channel.

[0036] Further, the shortest distance connecting the geometric center of the cross-section of the sample channel to the edge is less than 5000 μm, such as 30 nm - 5000 μm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 1000 nm, 2 μm, 5 μm, 10 μm, 20 μm, 50 μm, 100 μm, 200 μm, 500 μm, 1000 μm, 2000 μm, 5000 μm.

[0037] Further, the height of the micro-nano structure provided inside the sample channel is less than the shortest distance connecting the geometric center of the cross-section of the sample channel to the edge, and further is 10% - 90% of the shortest distance connecting the geometric center of the cross-section of the sample channel to the edge. For example, it is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%.

[0038] Further, the shortest distance connecting the edges through the geometric center of the cross-section of the microchannel is less than 2000 μm, such as 30 nm - 2000 μm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 1000 nm, 2 μm, 5 μm, 10 μm, 20 μm, 50 μm, 100 μm, 200 μm, 500 μm, 1000 μm, 2000 μm.

[0039] Further, the height of the micro-nano structure inside the microchannel is less than the shortest distance connecting the edges through the geometric center of the cross-section of the microchannel, and further is 10% - 90% of the shortest distance connecting the edges through the geometric center of the cross-section of the microchannel. For example, it is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%.

[0040] Further, the shortest distance connecting the edges through the geometric center of the cross-section of the compression channel is set to be 0.1% - 50% of the shortest distance connecting the edges through the geometric center of the cross-section of the sample channel. For example, it is 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 1%, 5%, 10%, 20%.

[0041] Further, the shortest distance connecting the edges through the geometric center of the cross-section of the compression channel is set to be 20% - 1000% of the size of the biological particle; and the distance of the longest straight line among the straight lines perpendicular to the straight line where the shortest distance on the cross-section of the compression channel is located is greater than or equal to the size of the biological particle, for example: 30 nm - 1000 μm.

[0042] Further, the distance of the longest straight line among the straight lines perpendicular to the straight line where the shortest distance on the cross-section of the compression channel is located is 30 nm - 1000 μm, such as 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 1000 nm, 2 μm, 5 μm, 10 μm, 20 μm, 50 μm, 100 μm, 200 μm, 500 μm, 1000 μm.

[0043] Further, the high-throughput micro-nano fluid control device is made of one or more materials among glass, quartz, silicon wafer, polydimethylsiloxane, silicon nitride wafer, silicon dioxide wafer, silicon carbide wafer and silicon-on-insulator.

[0044] The second aspect of the present invention provides a micro-nano fluid loading system, which includes the above-mentioned high-throughput micro-nano fluid control device, a first input module, a first flow control module, a first recovery module, and connection components for connecting between the modules;

[0045] The first input module is connected to the sample channel inlet in the high-throughput micro-nano fluid control device;

[0046] The first recovery module is connected to the ports of the microchannel array in the high-throughput micro-nano fluid control device;

[0047] The first flow control module is a module for controlling the opening and closing of the flow control outlet, adjusting the output flow rate of the sample solution at the flow control outlet, adjusting the input flow rate of the sample solution at the first input module, and controlling the start and stop of the first recovery module.

[0048] The first input module is used to input the sample solution. Especially under the adjustment of the first flow control module, the sample solution is input according to the set flow rate.

[0049] The first recovery module is used to recover the sample solution. Especially under the adjustment of the first flow control module, the sample solution is recovered according to the set flow rate.

[0050] In the third aspect of the present invention, a micro-nano fluid mixing system is provided. The micro-nano fluid mixing system includes the above-mentioned high-throughput micro-nano fluid control device, a second input module, a second flow control module, a second circulation module, and connection components for realizing the connection between each module;

[0051] The second input module is connected to the ports of the microchannel array in the high-throughput micro-nano fluid control device;

[0052] The second flow control module is a module for controlling the opening and closing of the flow outlet, adjusting the output flow rate of the sample solution at the flow control outlet, controlling the liquid at the flow control outlet to circulate back to the inlet of the sample channel through the second circulation module or discharging the liquid at the flow control outlet, and adjusting the input flow rate of the sample solution at the second input module;

[0053] The second circulation module communicates with the sample channel inlet and the flow control outlet, so that the sample solution can circulate in the sample channel.

[0054] The second input module is used to input the sample solution. Especially under the adjustment of the second flow control module, the sample is input according to the set flow rate.

[0055] In the fourth aspect of the present invention, a micro-nano fluid sorting system is provided. The micro-nano fluid sorting system includes the above-mentioned high-throughput micro-nano fluid control device, a third recovery module, a third flow control module, a third circulation module, and connection components for realizing the connection between each module;

[0056] The third recovery module is connected to the ports of the microchannel array in the high-throughput micro-nano fluid control device;

[0057] The third flow control module is a module that controls the opening and closing of the flow control outlet, adjusts the output flow rate of the sample solution at the flow control outlet, controls the liquid at the flow control outlet to circulate back to the inlet of the sample channel through the third circulation module or discharges the liquid at the flow control outlet, and controls the start and stop of the recovery module.

[0058] The third circulation module is connected to the inlet of the sample channel and the flow control outlet, so that the sample solution can circulate in the sample channel.

[0059] The third recovery module is used to recover the sample. Especially under the adjustment of the third flow control module, the sample solution is recovered according to the set flow rate.

[0060] Furthermore, the high-throughput micro-nano fluid control device contains at least two groups of microchannel arrays.

[0061] The fifth aspect of the present invention provides a use of the above high-throughput micro-nano fluid control device, which is used for loading exogenous therapeutic substances into biological particles, preparing liposomes, mixing samples, and sorting biological particles.

[0062] The sixth aspect of the present invention provides a use of the above micro-nano fluid loading system, which is used for loading exogenous therapeutic substances into biological particles.

[0063] The seventh aspect of the present invention provides a use of the above micro-nano fluid mixing system, which is used for preparing liposomes and mixing samples.

[0064] The eighth aspect of the present invention provides a use of the above micro-nano fluid sorting system, which is used for sorting biological particles.

[0065] The ninth aspect of the present invention provides a method for loading exogenous therapeutic substances into biological particles, and the method includes the following steps:

[0066] S01) Provide the above micro-nano fluid loading system, and prepare a mixed solution of biological particles and exogenous therapeutic substances;

[0067] S02) Fill the sample channel with a solvent and discharge the bubbles in the sample channel; then input the mixed solution into the sample channel of the high-throughput micro-nano fluid control device through the first input module. After the mixed solution fills the entire sample channel, close the flow control outlet of the sample channel;

[0068] S03) Continue to input the mixed solution. The mixed solution flows into the microchannel through the compression channel connected to the sample channel. After the biological particles are processed by the compression channel, the exogenous therapeutic substances are loaded into the interior of the biological particles, and a solution of biological particles loaded with exogenous therapeutic substances is obtained.

[0069] Further, the step S01) further includes a step of cleaning the microfluidic loading system.

[0070] Further, the step S03) further includes a step of recovering the solution of the biological particles loaded with the exogenous therapeutic substance by using the first recovery module and further purifying it.

[0071] Further, the purification step includes purifying by using methods such as ultracentrifugation, density gradient centrifugation, filtration, immunocapture, precipitation kit, size exclusion chromatography separation method, microfluidic separation method or polymer precipitation separation method to remove the unencapsulated exogenous therapeutic substance.

[0072] The tenth aspect of the present invention provides a method for preparing liposomes, and the preparation method includes the following steps:

[0073] S11) Provide the above-mentioned microfluidic mixing system; prepare an organic phase and an aqueous phase, wherein the organic phase contains substances for generating liposome membrane materials and a solvent capable of dissolving the substances for generating liposome membrane materials; the aqueous phase uses water or an aqueous salt solution of water as a solvent; the exogenous therapeutic substance is included in at least one of the aqueous phase and the organic phase;

[0074] S12) Inject the organic phase and the aqueous phase into the ports of the microchannels through the second input module respectively; the organic phase and the aqueous phase enter the sample channel through the compression channel for mixing;

[0075] S13) Continuously input the organic phase and the aqueous phase, and turn on the second circulation module so that the mixed solution of the organic phase and the aqueous phase can flow and mix and self-assemble in the sample channel to obtain liposomes loaded with exogenous therapeutic substances.

[0076] Further, in the step S12), the organic phase and the aqueous phase are respectively injected into the ports of different microchannel arrays through the second input module.

[0077] Further, the step S13) further includes a step of taking out the liposomes loaded with exogenous therapeutic substances from the flow control outlet and purifying them.

[0078] Further, the purification step includes purifying by using methods such as ultracentrifugation, density gradient centrifugation, filtration, immunocapture, precipitation kit, size exclusion chromatography separation method, microfluidic separation method or polymer precipitation separation method to remove the unencapsulated exogenous therapeutic substance.

[0079] The eleventh aspect of the present invention provides a method for sorting biological particles, and the sorting method includes the following steps:

[0080] S21) Provide the above-mentioned microfluidic sorting system, and prepare the biological particles to be sorted into a solution to be sorted;

[0081] S22) Input the liquid to be sorted into the sample channel of the high-throughput micro-nano fluidic control device through the third input module, and through the cascade connection between the third flow control module and the third circulation module, enable the liquid to be sorted to circulate in the sample channel; the biological particles are sorted according to their particle sizes in the compression channel, and the biological particles passing through the compression channel finally enter the microchannel array;

[0082] S23) Collect the biological particles in the microchannel array and the biological particles in the sample channel.

[0083] Further, in S21), the high-throughput micro-nano fluidic control device includes at least two groups of microchannel arrays, and the diameters of the compression channels connected to the sample channel and different microchannel arrays are different.

[0084] Further, in S23), it also includes the step of further concentrating and purifying the sorted biological particles.

[0085] The present invention utilizes micro-nano fluidic technology to design a high-throughput micro-nano fluidic control device with a microchannel system, compression channels, and sample channels, aiming to accurately manipulate and process biological particles such as extracellular vesicles at the nano-scale with high throughput, achieve reversible perforation of biological particles such as extracellular vesicles and loading of exogenous substances, thereby efficiently, uniformly, and stably preparing drug delivery carriers, and solving technical problems such as bubble problems, channel blockage, sample dead volume, and delivery carrier contamination caused by the huge size difference between microchannels and compression channels in existing micro-nano chips. Therefore, the present invention is significantly superior to the prior art in three aspects: the unique structural design of the device, the processing and manufacturing method, and the preparation of drug delivery carriers:

[0086] (1) The structural design of the high-throughput micro-nano fluidic control device is unique. By setting a sample channel between the microchannel array and the compression channel array, a sample solution input buffer area is formed between the microchannel array and the compression channel array. Through optimizing the preparation process and adjusting the flow control outlet, technical problems such as sample aggregation, channel blockage, bubble problems, and sample dead volume caused by the huge size difference between microchannels and compression channels in existing micro-nano chips can be effectively solved.

[0087] (2) The sample channel is further integrated with micro-nano structures, enabling the sample solution to be fully mixed when flowing in the sample channel, ensuring that the preparation process of the drug delivery carrier is more stable and reliable. In addition, the compression channel is integrated with nano-structures. When extracellular vesicles pass through the compression channel, a controllable mechanical effect can be exerted on biological particles such as extracellular vesicles, thereby precisely regulating the preparation process of the drug delivery carrier.

[0088] (3) The structural design of the high-throughput micro-nano fluidic control device is scalable. Not only can the number, shape, and number of layers of the microchannel array, sample channels, and compression channels be designed according to the processing throughput requirements of the sample solution, but also the number and shape of the sample channels can be designed according to the types of sample solutions, the mixing requirements of sample solutions, and the internal flow resistance distribution in the channels, not limited to Figure 2 the shape of the sample channels in

[0089] (4) The device processing and manufacturing method is compatible with traditional micro-nano processing technologies. The manufacturing method is mature and simple. The manufacturing methods of microchannels, compression channels, and sample channels include soft lithography, dry etching, wet etching, nanoimprinting, and 3D printing, etc. The bonding between the two can also be achieved through methods such as oxygen plasma bonding, silicon-silicon bonding, anodic bonding, and fusion bonding.

[0090] (5) The high-throughput micro-nano fluidic control device is universal for processing various types of biological particles and exogenous therapeutic substances. After precise processing by the device, different types of extracellular vesicles, subcellular structures with membrane structures, membrane vesicles secreted by microorganisms, cell membrane nanoparticles, artificially synthesized nanoparticles encapsulated by phospholipid bilayer membranes, liposomes, or virus vectors and other biological particles can be loaded with one or more exogenous therapeutic substances at the same time. Among them, exogenous therapeutic substances generally refer to biological small molecules, biological macromolecules, and nanomaterials with a size less than or equal to 200 nm. In addition, due to the sample channels with micro-nano structures provided in the high-throughput micro-nano fluidic control device, the device can also be used to synthesize liposomes, or synthesize cell membrane fragments into cell membrane nanoparticles, and with the help of the compression channels with nano-structures, through precise processing and mechanical action, sort or synthesize liposomes or cell membrane nanoparticles with specific sizes.

[0091] Based on micro-nano fluidic technology, a high-throughput micro-nano fluidic control device with characteristic dimensions matching the sizes of biological particles is constructed. By further setting sample channels in the microchannel / compression channel system formed by the microchannel array and the compression channel array, it is not only possible to precisely manipulate and process biological particles at the nanoscale with high throughput, meet the requirement of efficient loading of exogenous therapeutic substances by biological particles, but also effectively solve technical problems such as bubble problems, channel blockage, and sample dead volume caused by the huge size difference between microchannels and compression channels in the actual application of existing micro-nano chips. Through the structural design and channel layout of the present invention, not only can the sample processing throughput of the micro-nano fluidic control device be significantly improved, but also its operational flexibility in processing multi-scale complex biological particle samples can be enhanced, while avoiding problems such as contamination of the prepared products caused by residual dead volume, thus providing a new technical means for the efficient preparation of drug delivery carriers and the development of innovative drugs for biological particles.

[0092] Aiming at the technical drawbacks of the prior art, the present invention has the following beneficial effects:

[0093] (1) The present invention designs a new layout of a microchannel / compression channel system. By arranging a sample channel between the microchannel array and the compression channel array, a sample solution input buffer zone is formed between the microchannel array and the compression channel. The sample solution can be input into the compression channel through the sample channel and then reach the microchannel array. At the same time, since a flow control outlet is designed at the end of the sample channel, it can effectively prevent the sample from accumulating at the end of the microchannel, prevent the blockage and failure of the compression channel caused by sample retention, ensure that the micro-nano fluid control device can process the sample solution with high throughput, and help meet the requirements for a large sample size in animal experiments and clinical applications.

[0094] (2) The end of the sample channel designed in the present invention adopts a flow control outlet. After the sample solution enters the sample channel, the bubbles inside the channel can be smoothly discharged through the flow control outlet, ensuring that the sample solution can flow into the compression channel smoothly without being interfered by bubbles. In addition, due to the transportation of the sample solution through a catheter, there are often bubbles inside the solution. The flow control outlet design adopted by the sample channel can effectively discharge the bubbles inside the sample solution by opening or closing the outlet, further improving the reliability and stability of the micro-nano fluid control device.

[0095] (3) The present invention integrates refined micro-nano structures inside the sample channel, enabling the sample solution to be fully mixed when flowing through the sample channel. After the uniformly mixed sample solution enters the compression channel, since the ratio of extracellular vesicles to exogenous substances is consistent with the designed ratio, the preparation process of the drug delivery carrier is more stable and reliable. In addition, nano-structures are integrated inside the compression channel. When the extracellular vesicles pass through the compression channel, the nano-structures can exert controllable mechanical and fluid effects on biological particles such as extracellular vesicles, precisely regulating the preparation process of the drug delivery carrier, thereby improving the loading efficiency of biological particles such as extracellular vesicles for exogenous therapeutic substances.

[0096] (4) By arranging the sample channel in the present invention, the sample solution can directly enter the compression channel through the sample channel without passing through the inlet microchannel, thus effectively avoiding the pollution problem caused by the sample dead volume at the end of the inlet microchannel. At the same time, when designing the microchannel array, it is ensured that the end of the microchannel coincides with the compression channel, effectively avoiding the problem of sample solution dead volume in the microchannel, and further improving the quality and stability of the drug delivery carrier. Description of the Drawings

[0097] Figure 1 It is a flow chart of the technical solution for the design and application of a high-throughput micro-nano fluid control device.

[0098] Figure 2Structural diagram of one specific solution for a high-throughput micro-nano fluidic control device with a microchannel / compression channel system and a sample channel. Among them, a is a cross-sectional view in the corresponding direction of b1-b1' in Figure b, b is a top view of the high-throughput micro-nano fluidic control device with a microchannel / compression channel system and a sample channel, and c is a cross-sectional view in the corresponding direction of b2-b2' in Figure b.

[0099] Figure 3 Flow chart for fabricating a high-throughput micro-nano fluidic control device with a microchannel / compression channel system and a sample channel.

[0100] Figure 4 Schematic diagram of the processing and manufacturing process of a high-throughput micro-nano fluidic control device with a microchannel / compression channel system and a sample channel.

[0101] Figure 5 Flow chart for preparing a drug delivery carrier based on a high-throughput micro-nano fluidic control device.

[0102] Figure 6 Schematic diagram of a micro-nano fluidic loading system for preparing bio-particles loaded with exogenous therapeutic substances and the preparation of a drug delivery carrier. Among them, a is a schematic diagram of the assembly of a high-throughput micro-nano fluidic control device, and b is a schematic diagram of the principle of drug delivery carrier preparation. When bio-particles such as extracellular vesicles enter the compression channel through the sample channel, the compression channel with nanostructures exerts a mechanical effect on the bio-particles, stimulating the introduction of nanopores on their membrane surfaces, and then outputting through the microchannel.

[0103] Figure 7 Schematic diagram of the assembly of a micro-nano fluidic mixing system.

[0104] Figure 8 Schematic diagram of the assembly of a micro-nano fluidic sorting system.

[0105] Figure 9 Structural diagram of a high-throughput micro-nano fluidic control device with 3 groups of microchannel arrays in another embodiment. Among them, a is a cross-sectional view in the corresponding direction of b1-b1' in Figure b, b is a top view of the high-throughput micro-nano fluidic control device with a microchannel / compression channel system and a sample channel, and c is a cross-sectional view in the corresponding direction of b2-b2' in Figure b.

[0106] Figure 10 Structural diagram of a high-throughput micro-nano fluidic control device with 2 sample channels in another embodiment. Among them, a is a cross-sectional view in the corresponding direction of b1-b1' in Figure b, b is a top view of the high-throughput micro-nano fluidic control device with a microchannel / compression channel system and 2 sample channels, and c is a cross-sectional view in the corresponding direction of b2-b2' in Figure b. Specific implementation mode

[0107] It should be understood that the present invention is not limited to the details or methods set forth in the description or shown in the drawings. It should also be understood that the terms are for descriptive purposes only and should not be considered restrictive. In all the drawings, an effort has been made to use the same or similar reference numerals to denote the same or similar parts.

[0108] Some specific embodiments of the present invention provide a high-throughput micro-nano fluid control device, and the high-throughput micro-nano fluid control device includes a sample channel, a micro-channel array, and a compression channel array;

[0109] The sample channel is a channel that connects the inlet and the flow control outlet of the sample channel and is used to transport the sample;

[0110] The micro-channel array includes at least one micro-channel and ports connected to the micro-channel;

[0111] The compression channel array includes at least one independently arranged compression channel;

[0112] The micro-channel is connected to the sample channel through the compression channel;

[0113] The flow control outlet can be controllably opened or closed.

[0114] The ports connected to the micro-channel are inlets and outlets communicating with the outside world and can be used to inject samples into the compression channel or recover samples in the compression channel.

[0115] Independently arranged means that when the compression channel array includes multiple compression channels, each compression channel is not interconnected and is only used to connect the sample channel and the micro-channel.

[0116] In the present invention, the sample channel is a channel that connects the inlet of the sample channel and the flow control outlet and is used to transport the sample; wherein the flow control outlet can be controlled to be opened or closed; the sample can be injected through the inlet of the sample channel, and when bubbles appear or blockage occurs in the sample channel, the pressure, bubbles or blocked sample can be released by opening the flow control outlet; the cross-sectional area of the sample channel is larger than that of the compression channel; the cross-sectional shape of the sample channel is circular, square or rectangular, and the length of its diameter or length and width is greater than the size of the biological particles to be processed; the connection angle between the sample channel and the compression channel is greater than 0° and less than or equal to 90°, preferably 10°–90°; the sample channel communicates with multiple compression channels, and the compression channels can be arranged at any position of the sample channel, such as on the side wall, top or bottom; when multiple compression channels are provided, the compression channels are arranged in a symmetric arrangement or a staggered arrangement; the sample channel can include one flow control outlet or multiple flow control outlets; the cross-sectional areas of different segments in the sample channel can be the same or different, and when the cross-sectional areas are different, they can be used to regulate the flow resistance of different segments. For example, the part of the sample channel close to the inlet of the sample channel is wide, and the part close to the flow control outlet is narrow, so as to regulate the flow resistance of the sample channel and make it difficult for the sample solution to reach the end of the sample channel. The sample channel is one or more than two.

[0117] In the present invention, one end of the microchannel is connected to the microchannel port, and the other end is a closed structure; the cross-sectional shape of the microchannel is circular, square or rectangular, and the length of its diameter or length and width is greater than the size of the biological particles to be processed; micro-nano structures can be provided on the inner wall of the microchannel; the connection angle between the microchannel and the compression channel is greater than 0° and less than or equal to 90°, preferably 10°-90°; each microchannel communicates with multiple compression channels, and the compression channels can be arranged at any position of the microchannel, such as on the side wall, top or bottom; when multiple compression channels are provided, the compression channels are arranged in a symmetric arrangement or a staggered arrangement; the cross-sectional areas of different segments in the microchannel, or the cross-sectional areas of different microchannels in the same group of microchannels, can be the same or different, and when the cross-sectional areas are different, they can be used to regulate the flow resistance of different segments.

[0118] In the present invention, the microchannel array is at least one microchannel and a microchannel port connected to the microchannel; in a high-throughput micro-nano fluid control device, the microchannel array is one or more groups; when multiple groups of microchannel arrays are provided, different groups of microchannel arrays are not directly connected; preferably, multiple microchannels are provided; the microchannels are straight, curved, broken line, spiral, wavy, bifurcated, or other irregular shapes that meet the arrangement requirements; different microchannels in the same group of microchannel arrays can be distributed in any shape such as parallel distribution, concentric circle distribution, radial distribution, reticular distribution, etc., as long as the fluid therein can flow out or flow into through the microchannel port; the cross-sectional areas of different microchannels in the same group of microchannel arrays are the same or different, so as to control the output of the sample solution in the microchannel through different flow resistances formed by the microchannels.

[0119] In the present invention, the compression channel is a channel connecting the microchannel and the sample channel; the cross-sectional shape of the compression channel is not limited and can be circular, square, rectangular, oval, or irregular shape; the shortest distance from the geometric center of the cross-section of the compression channel to the edge is set to be 0.1% - 50% of the shortest distance from the geometric center of the cross-section of the sample channel to the edge; when processing biological particles, the shortest distance from the geometric center of the cross-section of the compression channel to the edge is set to be 20% - 1000% of the size of the biological particles; and the distance of the longest straight line among the straight lines perpendicular to the straight line where the shortest distance on the cross-section of the compression channel is located is greater than or equal to the size of the biological particles, for example: 30 nm - 1000 μm.

[0120] For example, when the cross-section of the compression channel is circular, the shortest distance from the geometric center of the cross-section of the compression channel to the edge is the diameter of the circle; when the cross-section of the compression channel is rectangular, the shortest distance from the geometric center of the cross-section of the compression channel to the edge is the short side length; when the cross-section of the compression channel is oval, the shortest distance from the geometric center of the cross-section of the compression channel to the edge is the length of the minor axis diameter; and the distance of the longest straight line among the straight lines perpendicular to the straight line where the shortest distance on the cross-section of the compression channel is located is greater than or equal to the size of the biological particles, for example: 30 nm - 1000 μm; nanostructures can be provided on the inner wall of the compression channel. The lengths of the compression channels can be the same or different, and the difference in the lengths of different compression channels does not exceed 50%. The cross-sectional shapes and areas of the compression channels provided at different positions are the same or different, and when the cross-sectional shapes and areas are different, the flow resistances generated are different. For example, at the front end position of the sample channel, that is, the compression channel near the entrance of the sample channel is wide, so the flow resistance is small and the sample solution is more easily transported into the compression channel; while at the rear end position of the sample channel, that is, the compression channel near the flow control outlet is narrow, so the flow resistance is large and the sample solution is less likely to be transported into the compression channel, avoiding the aggregation of the sample solution at the end of the sample channel.

[0121] In the present invention, the micro-nano structure is a surface structure at a scale of micrometers or smaller; the shape of the micro-nano structure can be any shape, such as: fishbone shape, rectangular shape, trapezoidal shape, triangular shape, circular shape, etc.; the arrangement of the micro-nano structure can be periodically arranged, and the form of the periodic arrangement can be any way, such as: linear arrangement, staggered arrangement, etc.; the micro-nano structure is disposed on the inner wall of the sample channel and / or the micro-channel, for example, disposed on the bottom or the side; the height of the micro-nano structure is 10% - 90% of the shortest distance connecting the center of the corresponding cross-section of the sample channel and / or the micro-channel to the edge, and the height of the sample channel and / or the micro-channel is the distance of the perpendicular intersection line of the plane where the micro-nano structure is located in the cross-section of the sample channel and / or the micro-channel. The micro-nano structure is a convex structure or a groove structure.

[0122] In the present invention, the nano-structure is a surface structure at a scale of nanometers or smaller; the shape of the nano-structure can be any shape, such as: fishbone shape, rectangular shape, trapezoidal shape, triangular shape, circular shape, etc.; the arrangement of the nano-structure can be periodically arranged, and the form of the periodic arrangement can be any way, such as: linear arrangement, staggered arrangement, etc.; the nano-structure is disposed on the inner surface of the compression channel, for example, disposed on the bottom or the side; the height of the nano-structure is 10% - 90% of the shortest distance connecting the center of the cross-section of the compression channel to the edge, and the height of the compression channel is the distance of the perpendicular intersection line of the plane where the nano-structure is located in the cross-section of the compression channel. The nano-structure is a convex structure or a groove structure.

[0123] In the present invention, the biological particles are any natural or artificial biological particles that can be used to load exogenous therapeutic substances. For example: cells, extracellular vesicles, liposomes, nanoparticles, polymer microspheres, virus particles, exosomes, microvesicles, apoptotic bodies, synthetic vesicles, nanovesicles, plant-derived vesicles (PDVs), syncytial vesicles, microalgae or algal particles.

[0124] In the present invention, the exogenous therapeutic substances refer to small molecule compounds, biological macromolecules, and nanomaterials, etc. with a size less than or equal to 200 nm. Small molecule compounds, such as: anticancer drugs such as doxorubicin, curcumin, and paclitaxel, biological molecules such as potassium ion probe molecules, calcium ion probe molecules, inositol trisphosphate, etc., biological macromolecules, such as: protein active components such as immunoglobulins, interleukins, bovine serum albumin, and endonucleases, nucleic acid active components such as plasmids, ribonucleic acids, and oligonucleotides, nanomaterials, such as: quantum dots, carbon nanotubes, and magnetic nanoparticles.

[0125] In the present invention, the material of the high-throughput micro-nano fluid control device can be any material that can meet the requirements of shape, size, and manufacturing process. For example, it can be made of one or more materials such as glass, quartz, silicon wafer, polydimethylsiloxane, silicon nitride wafer, silicon dioxide wafer, silicon carbide wafer, and silicon-on-insulator.

[0126] In the present invention, the number of layers of the high-throughput micro-nano fluid control device can be one layer or multiple layers. When the number of layers of the high-throughput micro-nano fluid control device is one layer, the sample channel, micro-channel array, and compression channel array are all arranged on the same layer. When the number of layers of the high-throughput micro-nano fluid control device is multiple layers, each layer can be provided with an independent sample channel, micro-channel array, and compression channel array; or the sample channel, micro-channel array, and compression channel array are arranged on different layers. For example, the sample channel is arranged on the upper layer, the micro-channel array is arranged on the lower layer, and the compression channel array connects the sample channel on the upper layer and the micro-channel on the lower layer; or the sample channel is arranged on the lower layer, the micro-channel array is arranged on the upper layer, and the compression channel array connects the sample channel on the lower layer and the micro-channel on the upper layer; or the sample channel and micro-channel array are both arranged in multiple layers, for example, the sample channel and micro-channel array are arranged in a spiral ascending or spiral descending manner.

[0127] In some specific technical solutions of the present invention, a micro-nano fluid loading system is also provided, which includes the above-mentioned high-throughput micro-nano fluid control device, a first input module, a first flow control module, a first recovery module, and connection components for realizing the connection between each module;

[0128] The first input module is connected to the sample channel inlet in the high-throughput micro-nano fluid control device;

[0129] The first recovery module is connected to the port of the micro-channel array in the high-throughput micro-nano fluid control device;

[0130] The first flow control module is a module for controlling the opening and closing of the flow control outlet, adjusting the output flow rate of the sample solution at the flow control outlet, adjusting the input flow rate of the sample solution at the first input module, and controlling the start and stop of the first recovery module.

[0131] The first input module is used to input samples. Especially under the adjustment of the first flow control module, the sample solution is input according to the set flow rate.

[0132] The first recovery module is used to recover samples. Especially under the adjustment of the first flow control module, the sample solution is recovered according to the set flow rate.

[0133] In some specific technical solutions of the present invention, a micro-nano fluid mixing system is further provided. The micro-nano fluid mixing system includes the above-mentioned high-throughput micro-nano fluid control device, a second input module, a second flow control module, a second circulation module, and connecting components for connecting between the modules;

[0134] The second input module is connected to the port of the microchannel array in the high-throughput micro-nano fluid control device;

[0135] The second flow control module is a module for controlling the opening and closing of the flow control outlet, adjusting the output flow rate of the sample solution at the flow control outlet, controlling the liquid at the flow control outlet to circulate back to the inlet of the sample channel through the second circulation module or discharging the liquid at the flow control outlet, and adjusting the input flow rate of the sample solution of the second input module;

[0136] The second circulation module communicates with the sample channel inlet and the flow control outlet, so that the sample solution can circulate in the sample channel.

[0137] The second input module is used to input the sample solution. Especially under the adjustment of the second flow control module, the sample solution is input into the microchannel array according to the set flow rate.

[0138] In some specific embodiments of the present invention, a micro-nano fluid sorting system is further provided. The micro-nano fluid sorting system includes the above-mentioned high-throughput micro-nano fluid control device, a third recovery module, a third flow control module, a third circulation module, and connecting components for connecting between the modules;

[0139] The third recovery module is connected to the port of the microchannel array in the high-throughput micro-nano fluid control device;

[0140] The third flow control module is a module for controlling the opening and closing of the flow control outlet, adjusting the output flow rate of the sample solution at the flow control outlet, controlling the liquid at the flow control outlet to circulate back to the inlet of the sample channel through the third circulation module or discharging the liquid at the flow control outlet, and controlling the start and stop of the third recovery module.

[0141] The third circulation module communicates with the sample channel inlet and the flow control outlet, so that the sample solution can circulate in the sample channel.

[0142] The third recovery module is used to recover the sample. Especially under the adjustment of the third flow control module, the sample in the microchannel array is recovered according to the set flow rate.

[0143] In some specific embodiments of the present invention, a method for loading exogenous therapeutic substances into biological particles is further provided. The method includes the following steps:

[0144] S01) Provide the above-mentioned micro-nano fluid loading system, formulate a mixed solution of biological particles and exogenous therapeutic substances; clean the micro-nano fluid loading system;

[0145] S02) Fill the sample channel with a solvent and discharge the bubbles in the sample channel; then input the mixed solution into the sample channel of the high-throughput micro-nano fluid control device through the input module. After the mixed solution fills the entire sample channel, close the flow control outlet of the sample channel;

[0146] S03) Continuously input the mixed solution. The mixed solution flows into the microchannel through the compression channel connected to the sample channel. The biological particles therein load the exogenous therapeutic substances into the interior of the biological particles after being processed by the compression channel, and obtain a solution of biological particles loaded with exogenous therapeutic substances;

[0147] Recover the solution of biological particles loaded with exogenous therapeutic substances by using the recovery module and further purify it.

[0148] The purification step includes purifying by using methods such as ultracentrifugation, density gradient centrifugation, filtration, immunocapture, precipitation kit, size exclusion chromatography separation method, micro-nano fluidic separation method or polymer precipitation separation method to remove the unencapsulated exogenous therapeutic substances.

[0149] Some specific embodiments of the present invention also provide a method for preparing liposomes, and the preparation method includes the following steps:

[0150] S11) Provide the above-mentioned micro-nano fluid mixing system; formulate an organic phase and an aqueous phase, wherein the organic phase contains substances for generating liposome membrane materials and a solvent capable of dissolving the substances for generating liposome membrane materials; the aqueous phase uses water or an aqueous salt solution of water as a solvent; at least one of the aqueous phase and the organic phase contains exogenous therapeutic substances;

[0151] S12) Inject the organic phase and the aqueous phase into the ports of the microchannel through the input module respectively, and inject the organic phase and the aqueous phase into the ports of different microchannel arrays through the input module respectively; the organic phase and the aqueous phase enter the sample channel through the compression channel for mixing;

[0152] S13) Continuously input the organic phase and the aqueous phase, and turn on the circulation module to enable the mixed solution of the organic phase and the aqueous phase to flow and mix and self-assemble in the sample channel, and obtain liposomes encapsulating exogenous therapeutic substances; the obtained liposomes encapsulating exogenous therapeutic substances are taken out from the flow control outlet and subjected to a purification step;

[0153] The purification step includes purifying by using methods such as ultracentrifugation, density gradient centrifugation, filtration, immunocapture, precipitation kit, size exclusion chromatography separation method, micro-nano fluidic separation method or polymer precipitation separation method to remove the unencapsulated exogenous therapeutic substances.

[0154] Some specific embodiments of the present invention also provide a method for sorting biological particles, and the sorting method includes the following steps:

[0155] S21) Provide the above-mentioned micro-nano fluid sorting system, and prepare the biological particles to be sorted into a liquid to be sorted; the high-throughput micro-nano fluid control device in the micro-nano fluid sorting system includes at least two groups of microchannel arrays, and the diameters of the compression channels connected to the sample channels and different microchannel arrays are different;

[0156] S22) Input the liquid to be sorted into the sample channel of the high-throughput micro-nano fluid control device through the input module, and through the cascade connection between the flow control module and the circulation module, enable the liquid to be sorted to circulate in the sample channel; the biological particles are sorted according to their particle sizes in the compression channels, and the biological particles passing through the compression channels finally enter the microchannel arrays;

[0157] S23) Collect the biological particles in the microchannel arrays and the biological particles in the sample channels; further concentrate and purify the sorted biological particles.

[0158] Example 1 A micro-nano fluid loading system for loading exogenous therapeutic substances into biological particles, as well as preparation and application

[0159] The overall process of the design and application of a high-throughput micro-nano fluid control device includes four steps: device structure design, device processing and manufacturing, delivery carrier preparation, and sample analysis and characterization, as Figure 1 shown:

[0160] Step A, device structure design:

[0161] Figure 2 Sub-figure a is the front view of a high-throughput micro-nano fluid control device designed according to a specific technical solution of the present invention, which has a microchannel / compression channel system and a sample channel (for the cross-sectional view corresponding to b1-b1' in view b of Figure 2 ). The high-throughput micro-nano fluid control device is divided into upper and lower layers. The upper layer is a glass substrate with an inlet of the sample channel, a flow control outlet, a first port, and a second port. The lower layer is a silicon wafer substrate with a compression channel array, a sample channel, a first microchannel array, and a second microchannel array. Micro-nano structures are integrated inside the sample channel and the microchannel for sample solution mixing, and nano-structures are integrated inside the compression channel for precise processing of biological particles.

[0162] Figure 2Sub - figure b is the corresponding top view of the above - mentioned high - throughput micro - nano fluidic control device of the present invention. The round holes at the upper and lower ends of the high - throughput micro - nano fluidic control device are respectively the inlet of the sample channel and the flow - control outlet. The round holes at the left and right ends of the device are respectively the first port of the first micro - channel array and the second port of the second micro - channel array. The top of the silicon wafer substrate contains multiple compression channels, and the bottom surface of each compression channel integrates periodically arranged nano - structures. The compression channels are perpendicular to and connected to the sample channel and the micro - channels, and connect the sample channel and the micro - channels. The top of the silicon substrate contains a sample channel, and the head and tail of the sample channel are respectively connected to the inlet of the sample channel and the flow - control outlet. The bottom surface of the sample channel integrates periodically arranged micro - nano structures. The top of the silicon substrate also contains a first micro - channel array and a second micro - channel array. The first micro - channel array includes multiple micro - channels and a first port connected to the micro - channels. The second micro - channel array includes multiple micro - channels and a second port connected to the micro - channels. The sample channel, the first micro - channel array, and the second micro - channel array are alternately arranged to form an interdigitated layout. When the bottom of the glass substrate is bonded to the top of the silicon wafer substrate, the compression channel array can connect the sample channel to the micro - channels in the first micro - channel array or connect the sample channel to the micro - channels in the second micro - channel array. The sample solution can be input through the sample channel, transmitted through the compression channels, and reach the first micro - channel array and the second micro - channel array respectively, and finally be output through the first port and the second port; or it can also be input through the first port or the second port, pass through the compression channels, and be output and collected through the flow - control outlet or the inlet of the sample channel of the sample channel.

[0163] Figure 2 Sub - figure c is the corresponding left view of the above - mentioned high - throughput micro - nano fluidic control device of the present invention (for Figure 2(cross-sectional view corresponding to b2-b2' in the b view). The compression channel connects the sample channel and the microchannels in the microchannel array. The bottom of the microchannel is provided with micro-nano structures, and the bottom of the compression channel is provided with nano structures. The cross-sections of the sample channel and the microchannel are rectangular. The width of the compression channel at the top of the silicon substrate can be set to be greater than or equal to the size of biological particles such as extracellular vesicles according to the sample processing throughput requirements (for example: 30 nm - 1000 μm). The cross-section of the compression channel is rectangular, and the height of the compression channel is set to 20% - 1000% of the size of biological particles such as extracellular vesicles to facilitate the processing of different types and sizes of biological particles such as extracellular vesicles; the height of the nano structure on the bottom surface of each compression channel is less than the height of the compression channel (for example: 10% - 90% of the height of the compression channel), and the width is less than or equal to the width of the compression channel (for example: 10% - 100% of the width of the compression channel); for the sample channel at the top of the silicon substrate, its height and width are set to be greater than or equal to the size of biological particles such as extracellular vesicles (for example: 30 nm - 1000 μm), just meeting the requirements for the input of the sample solution. Moreover, the height and width of the sample channel at different positions can be designed to have different sizes to control the input of the sample solution in the sample channel through different flow resistances formed inside the sample channel; the height of the micro-nano structure on the bottom surface of the sample channel is less than the height of the sample channel (for example: 10% - 90% of the height of the sample channel), and the width is less than or equal to the width of the sample channel (for example: 10% - 100% of the width of the sample channel); for the first microchannel array and the second microchannel array at the top of the silicon substrate, their height and width are set to be greater than or equal to the size of biological particles such as extracellular vesicles (for example: 30 nm - 1000 μm), just meeting the requirements for the output of the sample solution. Moreover, the height and width of the first / second microchannel at different positions can be designed to have different sizes to control the output of the sample solution in the microchannel through different flow resistances formed by the microchannel.

[0164] In some other specific embodiments, the material selection of the high-throughput micro-nano fluid control device can be replaced, and the materials for the upper layer and the lower layer can both be or independently be selected from common micro-nano manufacturing materials such as glass, quartz, silicon wafers, polydimethylsiloxane (PDMS), silicon nitride wafers, silicon dioxide wafers, silicon carbide wafers, and silicon-on-insulator (SOI wafers).

[0165] In some other specific embodiments, the number of microchannel arrays can be replaced by 1 group, or more than 2 groups of microchannel arrays can be set. For example, 3 - 10 groups.

[0166] In some other specific embodiments, the number of sample channels can be replaced by 2, or more than 2 sample channels can be set. For example, 3 - 10.

[0167] In some other specific embodiments, the sample channel can be a channel with a constant cross-sectional area, or a channel with different cross-sectional areas at different positions. For example, the front end of the sample channel is wide and the rear end is narrow, so as to regulate the flow resistance of the sample channel and make it difficult for the sample solution to reach the end of the sample channel.

[0168] In some other specific embodiments, the cross-sectional areas of the compression channels at different positions are different, resulting in different flow resistances at different positions of the compression channels. For example, the compression channel at the front end position of the sample channel (i.e., near the entrance of the sample channel) is wide, so the flow resistance is small and the sample solution is more easily transported into the compression channel, while the compression channel at the rear end position of the sample channel (i.e., near the flow control outlet) is narrow, so the flow resistance is large and the sample solution is less likely to be transported into the compression channel, avoiding the accumulation of the sample solution at the end of the sample channel.

[0169] In some other specific embodiments, the positions of the compression channel array, the sample channel, the first microchannel array, and the second microchannel array can be replaced by being arranged in different layers. Or the device is provided with multiple layers, and each layer has an independent compression channel array, sample channel, first microchannel array, and second microchannel array. Or the device is provided with multiple layers, the sample channel and the microchannel array are independently arranged in different layers and are connected by the compression channel array. Or the device is provided with multiple layers, the sample channel and / or the microchannel array penetrate multiple layers, such as: spiraling upward, zigzagging upward, etc., and a connected compression channel array is provided in the sample channel and the microchannel array.

[0170] In some other specific embodiments, the high-throughput micro-nano fluid control device can be replaced by multiple layers, as long as there are a sufficient number of compression channel arrays connecting and communicating between any sample channel and the first / second microchannel arrays.

[0171] In some other specific embodiments, when the compression channel array connects the sample channel and the first / second microchannel arrays, it does not have to be 90°, and it can be connected at other angles, such as 1° - 89°.

[0172] In some other specific embodiments, the compression channels may not be parallel to each other. For example, they can be radially arranged, or arranged in an alternating pattern of acute and obtuse angles.

[0173] In some other specific embodiments, the relationship between the sample channel and the microchannel is not limited to being parallel, and the geometric relationship between the compression channel and the sample channel or the microchannel in the horizontal direction is not limited to being perpendicular, as long as the compression channel array can connect the sample channel and the microchannel.

[0174] In some other specific embodiments, the arrangement of multiple nanostructures is not restricted. For example: linear arrangement, staggered arrangement, etc. The shape of the nanostructures is not limited to fishbone shape, and can also be rectangle, trapezoid, triangle, circle, etc.

[0175] In some other specific embodiments, the arrangement of multiple micro-nano structures is not restricted. For example: linear arrangement, staggered arrangement, etc. The shape of the micro-nano structures is not limited to fishbone shape, and can also be rectangle, trapezoid, triangle, circle, etc.

[0176] Step B, device processing and fabrication:

[0177] The preparation of the above-mentioned high-throughput micro-nano fluid control device of the present invention can be prepared by conventional methods known in the prior art. For example, 3D printing or mask etching technology, soft lithography, dry etching, wet etching, nanoimprinting, etc.

[0178] In a specific implementation scheme, the processing and fabrication process includes three parts: the fabrication of the compression channel array layer, the fabrication of the sample channel and the micro-channel array layer, and device bonding, as Figure 3 shown. The main processing steps of the compression channel array layer include: the fabrication of the compression channel mask, the etching of the compression channel, the fabrication of the nanostructure mask, and the etching of the nanostructure, see Figure 3 sub-figure S1. The main processing steps of the sample channel and the micro-channel array layer include: the fabrication of the micro-channel mask, the etching of the micro-channel, the fabrication of the micro-nano structure mask, and the etching of the micro-nano structure, see Figure 3 sub-figure S2. The main processing steps of device bonding include: drilling holes in the glass substrate and bonding the glass substrate to the silicon wafer substrate, see Figure 2 sub-figure S3.

[0179] Step S1: Fabricate the compression channel array layer, which includes sub-steps B1–B4

[0180] Sub-step B1, fabrication of the compression channel mask:

[0181] Clean and dry the silicon wafer substrate, then hydrophilize the surface of the silicon wafer substrate and spin-coat a layer of adhesion promoter, then spin-coat a layer of positive photoresist on its surface and bake it, and finally perform exposure treatment using the compression channel mask plate, see Figure 4 sub-figure a; Immerse the exposed silicon wafer substrate in the developer for development, and perform hardening treatment at a certain temperature to further harden the photoresist, and then the compression channel mask can be obtained, see Figure 4 sub-figure b.

[0182] Sub-step B2, etching of the compression channel:

[0183] The silicon wafer substrate with a compression channel mask is subjected to reactive ion etching to etch a compression channel array on the top of the silicon wafer substrate, as shown in Figure 4 Neutron figure c; The etched silicon wafer substrate is immersed in an organic solvent or a stripping solution to remove the compression channel mask on the surface of the silicon wafer substrate, as shown in Figure 4 Neutron figure d.

[0184] Sub-step B3, nanostructure mask fabrication:

[0185] The silicon wafer substrate with a compression channel array is cleaned and dried, then the surface of the silicon wafer substrate is hydrophilized and a layer of adhesion promoter is spin-coated, then a layer of positive photoresist is spin-coated on its surface and baked, and finally alignment exposure is performed using a nanostructure mask plate, as shown in Figure 4 Neutron figure e; The exposed silicon wafer substrate is developed in a developer solution and subjected to a hard-bake treatment at a certain temperature to further harden the photoresist, and a nanostructure mask can be obtained, as shown in Figure 4 Neutron figure f.

[0186] Sub-step B4, nanostructure etching:

[0187] The silicon wafer substrate with a nanostructure mask is subjected to reactive ion etching to etch nanostructures on the bottom surface of the compression channels, as shown in Figure 4 Neutron figure g; The etched silicon wafer substrate is immersed in an organic solvent or a stripping solution to remove the nanostructure mask on the surface of the silicon wafer substrate, and a compression channel array layer with nanostructures can be obtained on the top of the silicon wafer substrate, as shown in Figure 4 Neutron figure h.

[0188] Step S2: Fabricate the sample channels and the microchannel array layer, and this step includes sub-steps B5–B8

[0189] Sub-step B5, microchannel mask fabrication:

[0190] The silicon wafer substrate with a compression channel array layer is cleaned and dried, then the surface of the silicon wafer substrate is hydrophilized and a layer of adhesion promoter is spin-coated, then a layer of positive photoresist is spin-coated on its surface and baked, and finally exposure treatment is performed using a microchannel mask plate, as shown in Figure 4 Neutron figure i; The exposed silicon wafer substrate is developed in a developer solution and subjected to a hard-bake treatment at a certain temperature to further harden the photoresist, and a microchannel mask can be obtained, as shown in Figure 4 Neutron figure j.

[0191] Sub-step B6, microchannel etching:

[0192] The silicon wafer substrate with a microchannel mask is subjected to deep silicon etching to etch the sample channels and the microchannel array on the top of the silicon wafer substrate, as shown in Figure 4Sub - figure k; Immerse the etched silicon wafer substrate in an organic solvent or a stripping solution to remove the microchannel mask on the surface of the silicon wafer substrate, as shown in Figure 4 Sub - figure l.

[0193] Sub - step B7, Fabrication of the micro - nano structure mask:

[0194] Clean and dry the silicon wafer substrate with sample channels and microchannel arrays, then hydrophilize the surface of the silicon wafer substrate and spin - coat an adhesion promoter, then spin - coat a positive photoresist on its surface and bake it. Finally, perform alignment exposure using a micro - nano structure mask plate, as shown in Figure 4 Sub - figure m; Immerse the exposed silicon wafer substrate in a developer for development and perform a hard - baking treatment at a certain temperature to further harden the photoresist, and then a micro - nano structure mask can be obtained, as shown in Figure 4 Sub - figure n.

[0195] Sub - step B8, Micro - nano structure etching:

[0196] Perform deep silicon etching on the silicon wafer substrate with the micro - nano structure mask, so as to etch micro - nano structures on the bottom surface of the microchannels, as shown in Figure 4 Sub - figure o; Immerse the etched silicon wafer substrate in an organic solvent or a stripping solution to remove the micro - nano structure mask on the surface of the silicon wafer substrate, and then dry the silicon wafer substrate, and a sample channel with micro - nano structures and a microchannel array layer can be obtained on the top of the silicon wafer substrate, as shown in Figure 4 Sub - figure p.

[0197] Step S3: Device bonding, this step includes sub - steps B9 - B10

[0198] Sub - step B9, Drilling holes in the glass substrate:

[0199] Use a laser to drill holes in specific positions of the glass substrate to facilitate the formation of the inlet, flow - control outlet, first port, and second port of the device, as shown in Figure 4 Sub - figure q.

[0200] Sub - step B10, Bonding the glass substrate and the silicon wafer substrate:

[0201] Clean the drilled glass substrate and clean the etched silicon wafer substrate, then align and bond the bottom of the glass substrate with the top of the silicon wafer substrate, and the fabrication of the high - throughput micro - nano fluid control device is completed, as shown in Figure 4 Sub - figure r.

[0202] In some other embodiments, device processing can be prepared by means of 3D printing, soft lithography, dry etching, wet etching, nano - imprinting, hot - embossing lithography, injection molding, laser micro - machining.

[0203] Step C, Preparation of Drug Delivery Carrier:

[0204] After the above-mentioned high-throughput micro-nano fluidic control device with a microchannel / compression channel system and a sample channel is processed and fabricated, experiments on the preparation of drug delivery carriers for biological particles such as extracellular vesicles can be further carried out. Therefore, this step will introduce in detail the usage methods and operation steps for the high-throughput micro-nano fluidic control device to achieve efficient, uniform, and stable preparation of drug delivery carriers. The experiments on the preparation of drug delivery carriers mainly include the assembly of the preparation system, the extraction of extracellular vesicles, the preparation of the sample solution, the treatment of the sample by the device, the preparation of the drug delivery carrier, and the collection and purification of the sample. The preparation process is as Figure 5 shown.

[0205] Sub-step C1, Preparation of Micro-nano Fluid Loading System:

[0206] After designing and fabricating a high-throughput micro-nano fluidic control device using micro-nano fluidic technology, the device needs to be sealed and assembled so that the inlet, flow control outlet, and ports of the sample channel are respectively connected to conduits, and the conduits are connected to the first input module, the first flow control module, and the first recovery module to obtain a micro-nano fluid sorting system. Then, subsequent experiments on the preparation of drug delivery carriers for biological particles such as extracellular vesicles are carried out, as shown in Figure 6 subfigure a in the middle.

[0207] Sub-step C2, Extraction of Extracellular Vesicles:

[0208] Before carrying out the experiments on the preparation of drug delivery carriers, biological particles such as extracellular vesicles also need to be extracted and separated from biological samples. Commonly used biological samples include cell culture supernatants, plasma, serum, urine, saliva, cerebrospinal fluid, ascites, amniotic fluid, semen, synovial fluid, bronchial fluid, tears, bile, gastric acid, lymph, pleural effusion, gastrointestinal lavage fluid, bronchoalveolar lavage fluid, milk, grapes, grapefruit, lemon, watermelon, carrots, ginger, tomatoes, broccoli, or ginseng, etc. The extraction methods include ultracentrifugation, density gradient centrifugation, filtration, immunocapture, precipitation kits, size exclusion chromatography separation methods, micro-nano fluidic separation methods, and polymer precipitation separation, etc., which are common methods for the extraction of extracellular vesicles. Finally, biological particles such as extracellular vesicles are resuspended in biological solutions such as phosphate buffer solution, cell culture medium, or physiological saline for later use.

[0209] Sub-step C3, Preparation of Sample Solution:

[0210] Mix a solution of biological particles such as extracellular vesicles with a certain concentration and a solution of exogenous therapeutic substances with a certain concentration. Due to the blocking effect of the membrane of biological particles such as extracellular vesicles, the substance molecules can only be distributed outside the extracellular vesicles, as shown in Figure 6Sub - figure b. In addition, the exogenous therapeutic substances proposed in the present invention generally refer to biomolecules, biopolymers, and nanomaterials with a size less than or equal to 200 nm, such as anticancer drugs like doxorubicin, curcumin, and paclitaxel; protein - based drug models such as immunoglobulins, interleukins, bovine serum albumin, and endonucleases; nanomaterial - based drug models such as quantum dots, carbon nanotubes, and magnetic nanoparticles; nucleic acid - based drug models such as plasmids, ribonucleic acids, and oligonucleotides; and biomolecules such as potassium ion probe molecules, calcium ion probe molecules, and inositol trisphosphate.

[0211] Sub - step C4, device - processing of the sample:

[0212] To avoid the generation of bubbles in the sample channel, first, the catheter and the sample channel are rinsed with deionized water, ultrapure water, or distilled water, etc., and then the bubbles generated in the sample channel are discharged. After the bubbles are completely discharged, the uniformly mixed sample solution is input through the input module and into the sample channel of the high - throughput micro - nano fluidic device via the catheter. When the sample solution fills the entire sample channel, the flow control outlet of the sample channel is then closed. Subsequently, after the sample solution is mixed through the sample channel and the micro - nano structure on the bottom surface of the sample channel, the uniformly mixed and proportion - consistent sample solution is transported to the compression channel array; with the help of the flow control outlet design at the end of the sample channel, it is also possible to effectively prevent the sample solution from accumulating at the end of the micro - channel, preventing problems such as clogging and failure of the compression channel caused by the retention of biological particles such as extracellular vesicles, and ensuring that the high - throughput micro - nano fluidic device can efficiently process the sample solution. When biological particles such as extracellular vesicles pass through the compression channel, they will be precisely processed by mechanical actions such as the mechanical compression force, fluid shear force, and wall friction force of the compression channel and the nano - structure on the bottom surface of the compression channel, disturbing the rearrangement of lipid molecules on the surface of the extracellular vesicle membrane and introducing nanoscale pores that exist transiently and do not damage the membrane structure, thereby providing a transport channel for the loading of exogenous therapeutic substances, see Figure 6 Sub - figure b.

[0213] Sub - step C5, preparation of drug - delivery carriers:

[0214] When biological particles such as extracellular vesicles move from the compression channel to the micro - channel, a large number of nanoscale pores will form on the membrane surface of the biological particles such as extracellular vesicles, and exogenous substance molecules can enter the biological particles such as extracellular vesicles through the nanoscale pores. When the biological particles such as extracellular vesicles are not subjected to mechanical action in the micro - channel, the nanoscale pores recover without damage due to the membrane fluidity effect, thus realizing the preparation of drug - delivery carriers of biological particles such as extracellular vesicles, see Figure 6 Sub - figure b.

[0215] Sub - step C6, sample collection and purification:

[0216] The sample solution processed by the high-throughput micro-nano fluid control device is collected through the recovery module, and then the extracellular vesicles and other biological particles in the sample solution are purified by methods such as ultracentrifugation, density gradient centrifugation, filtration, immunocapture, precipitation kits, size exclusion chromatography separation method, micro-nano fluidic separation method, and polymer precipitation separation, etc., to remove the excess exogenous therapeutic substances in the sample solution, and a drug delivery carrier with stable quality can be obtained.

[0217] Step D, sample analysis and characterization:

[0218] After being processed by the high-throughput micro-nano fluid control device and obtaining the drug delivery carrier, the formation and recovery of nanopores can be observed through tools such as cryo-electron microscopy, transmission electron microscopy, and atomic force microscopy. The physical morphology of extracellular vesicles and other biological particles, the loading efficiency of extracellular vesicles and other biological particles, the dose of loaded exogenous therapeutic substances, the zeta potential of extracellular vesicles and other biological particles, etc. can be characterized by analysis methods such as nano-flow cytometry analyzer, flow cytometer, super-resolution optical imaging system, nanoparticle tracking analyzer, fluorescent dye kit, real-time fluorescence quantitative polymerase chain reaction instrument, mass spectrometer, spectrophotometer, etc. In addition, the integrity and functionality of the drug delivery carrier of extracellular vesicles and other biological particles can be evaluated through in vitro cell experiments and animal experiments, so as to verify the application potential of the high-throughput micro-nano fluid control device in the fields of biomedical research and clinical translation practice, etc.

[0219] Example 2 A micro-nano fluid mixing system and a method for preparing liposomes using the system

[0220] (1) Synthesis system assembly

[0221] Based on the same high-throughput micro-nano fluid control device as in the above example, a micro-nano fluid mixing system is assembled. By connecting the ports of the high-throughput micro-nano fluid control device, the sample channel inlet, and the flow control outlet to the catheter respectively, and connecting the catheter to the second input module, the second circulation module, and the second flow control module, a micro-nano fluid mixing system is obtained, so as to provide support for the synthesis and preparation of biological particles such as liposomes in the follow-up;

[0222] Among them, the second input module is respectively connected to the port;

[0223] The second circulation module connects the sample channel inlet and the flow control outlet, so that the liquid can circulate in the sample channel;

[0224] The second flow control module controls the opening and closing of the flow outlet, adjusts the output flow rate of the sample solution at the flow control outlet, controls the liquid at the flow control outlet to circulate back to the inlet of the sample channel through the second circulation module or discharges the liquid at the flow control outlet, and adjusts the input flow rate of the sample solution of the second input module.

[0225] (2) Preparation of sample solution

[0226] During the synthesis of biological particles such as liposomes, an organic phase solution with a certain concentration and an aqueous phase solution with a certain concentration need to be prepared. Commonly used organic phase solutions include lipid substances such as lecithin and cholesterol, which are dissolved in organic solvents (such as chloroform, vinyl chloride, ethanol, etc.). The aqueous phase solution includes exogenous therapeutic substances and is dissolved in solvents such as normal saline, buffer solution, deionized water, and PBS buffer solution. If the exogenous therapeutic substance is easily soluble in organic solvents and not easily soluble in water, the exogenous therapeutic substance can also be dissolved in the organic phase, and the aqueous phase only uses aqueous solvents, such as normal saline, buffer solution, deionized water, PBS buffer solution, etc.

[0227] (3) Device treatment of samples

[0228] Using the second input module, the organic phase solution is input into the microchannel array through the port, and through the mixing effect of the micro-nano structure at the bottom of the microchannel, the organic phase solution is evenly distributed into the compression channel array; similarly, the second input module inputs the aqueous phase solution into the microchannel array through the port, and through the mixing effect of the micro-nano structure at the bottom of the microchannel, the aqueous phase solution is evenly distributed into the compression channel array. When the organic phase solution and the aqueous phase solution pass through the compression channel array and meet in the sample channel, the hydrodynamic effect can ensure the high-speed mixing of the two solutions. After combining with the mixing effect of the micro-nano structure on the bottom surface of the sample channel, the self-assembly process of lipid molecules in the aqueous phase can be precisely regulated.

[0229] In addition, the input flow rate ratio of the two solutions is matched according to the type and properties of the required biological particles such as liposomes to ensure the stability and drug-loading capacity of the liposomes during the synthesis process.

[0230] (4) Preparation of liposome particles

[0231] During the liposome synthesis process, the organic phase solution and the aqueous phase solution are mixed at high speed and refined in the micro-compression channel. With the help of the emulsification effect, lipid molecules are promoted to assemble a bilayer structure in the aqueous phase, and finally liposome particles are formed. This process can precisely control the flow rates of the two solutions, the intensity of the fluid action, and the pressure in the channel by regulating the microchannel, compression channel, and their micro-nano structure design, thereby effectively regulating the size and distribution of liposome particles.

[0232] (5) Sample collection and purification

[0233] After the liposomes are synthesized, the sample solution processed in the sample channel is collected using a flow control module, and then the liposomes are purified by methods such as size exclusion chromatography, immunocapture, ultracentrifugation, density gradient centrifugation, precipitation kits, filtration, micro-nano fluidic separation methods, and polymer precipitation separation to remove excess solvents, unreacted raw materials, and exogenous therapeutic substances, and finally liposomes with stable quality and uniformity are obtained as drug delivery carriers.

[0234] Example 3 A micro-nano fluid sorting system and a method for sorting biological particles using the system

[0235] (1) Assembly of the sorting system

[0236] Based on the same high-throughput micro-nano fluid control device as in the above example, the sorting system for biological particles such as extracellular vesicles is assembled. By connecting the sample channel inlet, flow control outlet, and ports to the catheter respectively, and connecting the catheter to the third circulation module, the third flow control module, and the third recovery module, a micro-nano fluid mixing system is obtained.

[0237] Among them, the third recovery module communicates with the microchannel port in the high-throughput micro-nano fluid control device;

[0238] The third flow control module is a module that controls the opening and closing of the flow outlet, adjusts the output flow rate of the sample solution at the flow control outlet, controls the liquid at the flow control outlet to circulate back to the inlet of the sample channel through the third circulation module or discharges the liquid at the flow control outlet, and controls the start and stop of the recovery module;

[0239] The third circulation module communicates with the sample channel inlet and the flow control outlet, enabling the liquid to circulate in the sample channel.

[0240] Ensure that the system can smoothly perform subsequent sorting operations on biological particles such as extracellular vesicles.

[0241] (2) Preparation of the sample solution

[0242] Before sorting biological particles such as extracellular vesicles, it is necessary to prepare a sample solution containing the extracellular vesicles or other biological particles to be sorted. Common sources of the sample solution include cell culture supernatants, plasma, serum, urine, saliva, cerebrospinal fluid, ascites, amniotic fluid, semen, synovial fluid, bronchial fluid, tears, bile, gastric acid, lymph, pleural effusion, gastrointestinal lavage fluid, bronchoalveolar lavage fluid, milk, grapes, grapefruit, lemon, watermelon, carrots, ginger, tomatoes, broccoli, or ginseng, etc., or a biological particle solution extracted from the original biological sample by methods such as ultracentrifugation, density gradient centrifugation, filtration, immunocapture, precipitation kits, size exclusion chromatography separation methods, micro-nano fluidic separation methods, and polymer precipitation separation. In addition, the concentration and volume of the sample solution should be adjusted according to the sorting requirements to ensure the fluidity and sorting efficiency of the biological particles in the micro-compression channel.

[0243] (3) Device processing of the sample

[0244] Use the third circulation module to input the sample solution into the sample channel in the high-throughput micro-nano fluidic control device through the sample channel inlet, and through the cascade connection between the third flow control module and the third circulation module, the sample solution can circulate in the sample channel. In the sample channel, the flow rate of the sample solution is adjusted through the flow control outlet to ensure that biological particles such as extracellular vesicles are sorted according to their particle sizes in the compression channel and finally reach the microchannel array, and then can be collected by the third recovery module to achieve the sorting of biological particles of a specific size.

[0245] (4) Sorting of biological particles

[0246] During the sorting process of biological particles such as extracellular vesicles, biological particles such as extracellular vesicles with a size less than or equal to the height of the compression channel can enter the microchannel through the size sorting of the compression channel array, while biological particles with a size greater than the height of the compression channel cannot pass through the compression channel due to the size limitation of the compression channel array and can only circulate or be discharged in the sample channel. In addition, by adjusting the flow rate of the sample solution through the flow control outlet, it is possible to effectively avoid the blockage of the compression channel array by biological particles such as extracellular vesicles with a larger size. Further, a reverse flow rate can be applied in the microchannel array through the third recovery module to effectively remove large-size biological particles in the compression channel array and discharge them smoothly through the flow control outlet.

[0247] (5) Sample collection

[0248] After being processed by the high-throughput micro-nano fluidic control device and collected by the third recovery module, the efficient sorting of biological particles such as extracellular vesicles can be achieved, so as to obtain biological particles of a specific size as drug delivery carriers. The sorted biological particles can be further concentrated or used for subsequent experiments.

Claims

1. A high-throughput micro-nano fluid control device, characterized in that The high-throughput micro-nano fluid control device includes a sample channel, a micro-channel array, and a compression channel array; The sample channel is a channel that connects the inlet and the flow control outlet of the sample channel and is used to transport the sample; The micro-channel array includes at least one micro-channel and ports connected to the micro-channel; The compression channel array includes at least one compression channel; The micro-channel is connected to the sample channel through the compression channel; The flow control outlet can be controllably opened or closed; Preferably, the high-throughput micro-nano fluid control device includes at least one group of micro-channel arrays; Preferably, the micro-channel array includes two or more micro-channels; Preferably, the high-throughput micro-nano fluid control device includes at least one sample channel; Preferably, the sample channel is a channel that includes one or more inlets communicating with the sample channel and one or more flow control outlets; Preferably, each of the micro-channels is connected to the sample channel through at least one compression channel; Preferably, the high-throughput micro-nano fluid control device is at least one layer; Preferably, when multiple layers of the high-throughput micro-nano fluid control device are provided, each layer can be separately provided with a sample channel, a micro-channel array, and a compression channel; Preferably, micro-nano structures are provided in the sample channel; Preferably, micro-nano structures are provided in the micro-channel; Preferably, nano-structures are provided in the compression channel; Preferably, the inlet and the flow control outlet of the sample channel are connected through a circulation module, and the circulation module is used to circulate the sample flowing out of the flow control outlet back into the sample channel through the inlet of the sample channel.

2. A micro-nano fluid loading system, characterized in that, The micro-nano fluid loading system includes the high-throughput micro-nano fluid control device according to claim 1, a first input module, a first flow control module, a first recovery module, and connection components for connecting between the modules; The first input module is communicated with the inlet of the sample channel in the high-throughput micro-nano fluid control device; The first recovery module is communicated with the ports of the micro-channel array in the high-throughput micro-nano fluid control device; The first flow control module is a module for controlling the opening and closing of the flow control outlet, adjusting the output flow rate of the sample solution at the flow control outlet, adjusting the input flow rate of the sample solution at the first input module, and controlling the start and stop of the first recovery module; The first input module is used to input the sample solution; The first recovery module is used to recover the sample solution.

3. A micro-nano fluid mixing system, characterized in that, The micro-nano fluid mixing system includes the high-throughput micro-nano fluid control device according to claim 1, a second input module, a second flow control module, a second circulation module, and connection components for connecting between the modules; The second input module is communicated with the ports of the micro-channel array in the high-throughput micro-nano fluid control device; The second flow control module is a module for controlling the opening and closing of the flow outlet, adjusting the output flow rate of the sample solution at the flow control outlet, controlling the liquid at the flow control outlet to be circulated back to the inlet of the sample channel through the second circulation module or discharging the liquid at the flow control outlet, and adjusting the input flow rate of the sample solution at the second input module; The second circulation module communicates with the sample channel inlet and the flow control outlet, so that the sample solution can circulate in the sample channel; The second input module is used to input a sample solution.

4. A micro-nano fluid sorting system, characterized in that, The micro-nano fluid sorting system includes the high-throughput micro-nano fluid control device, the third recovery module, the third flow control module, the third circulation module, and connection components for connecting between the modules as described in claim 1; The third recovery module is communicated with the port of the microchannel array in the high-throughput micro-nano fluid control device; The third flow control module is a module for controlling the opening and closing of the flow control outlet, adjusting the output flow rate of the sample solution at the flow control outlet, controlling the liquid at the flow control outlet to circulate back to the inlet of the sample channel through the third circulation module or discharging the liquid at the flow control outlet, and controlling the start and stop of the recovery module; The third circulation module communicates the sample channel inlet and the flow control outlet, so that the sample solution can circulate in the sample channel; The third recovery module is used to recover the sample solution; Preferably, the high-throughput micro-nano fluid control device contains at least two groups of microchannel arrays.

5. Use of the above-mentioned high-throughput micro-nano fluid control device, characterized in that, It is used for loading exogenous therapeutic substances into biological particles, liposome preparation, sample mixing, and biological particle sorting.

6. The use of the micro-nano fluid loading system according to claim 3, which is used for loading exogenous therapeutic substances into biological particles.

7. Use of the micro-nano fluid mixing system according to claim 4, characterized in that, It is used for liposome preparation and sample mixing.

8. Use of the micro-nano fluid sorting system according to claim 5, characterized in that, It is used for biological particle sorting.

9. A method for loading exogenous therapeutic substances into biological particles, characterized in that, The method includes the following steps: S01) Provide the micro-nano fluid loading system according to claim 3, and prepare a mixture of biological particles and exogenous therapeutic substances; S02) Fill the sample channel with a solvent and discharge the air bubbles in the sample channel; then input the mixture into the sample channel of the high-throughput micro-nano fluid control device through the first input module. After the mixture fills the entire sample channel, close the flow control outlet of the sample channel; S03) Continuously input the mixture, and the mixture flows into the microchannel through the compression channel connected to the sample channel. After the biological particles are processed by the compression channel, the exogenous therapeutic substances are loaded into the interior of the biological particles, and a solution of biological particles loaded with exogenous therapeutic substances is obtained; Preferably, step S01) further includes the step of cleaning the micro-nano fluid loading system; Preferably, step S03) further includes the step of recovering and further purifying the solution of biological particles loaded with exogenous therapeutic substances by using the first recovery module; Preferably, the purification step includes purification by using methods such as ultracentrifugation, density gradient centrifugation, filtration, immunocapture, precipitation kit, size exclusion chromatography separation method, micro-nano fluidic separation method, or polymer precipitation separation to remove the unencapsulated exogenous therapeutic substances.

10. A method for preparing liposomes, characterized in that, The preparation method includes the following steps: S11) Provide the micro-nano fluid mixing system according to claim 4; prepare an organic phase and an aqueous phase, wherein the organic phase contains substances for forming the liposome membrane material and a solvent capable of dissolving the substances for forming the liposome membrane material; the aqueous phase uses water or an aqueous salt solution as the solvent; at least one of the aqueous phase and the organic phase contains exogenous therapeutic substances; S12) Inject the organic phase and the aqueous phase into the ports of the microchannel through the second input module respectively; the organic phase and the aqueous phase enter the sample channel through the compression channel for mixing; S13) Continuously input the organic phase and the aqueous phase, and activate the second circulation module so that the mixture of the organic phase and the aqueous phase can flow and mix in the sample channel and self-assemble to obtain liposomes encapsulating exogenous therapeutic substances; Preferably, in step S12), the organic phase and the aqueous phase are respectively injected into the ports of different microchannel arrays through the second input module; Preferably, step S13) further includes the step of taking out the liposomes encapsulating exogenous therapeutic substances obtained from the flow control outlet and purifying them; Preferably, the purification step includes purifying by using methods such as ultracentrifugation, density gradient centrifugation, filtration, immunocapture, precipitation kits, size exclusion chromatography separation methods, micro-nano fluidic separation methods or polymer precipitation separation to remove the unencapsulated exogenous therapeutic substances.

11. A sorting method for biological particles, characterized in that, The sorting method includes the following steps: S21) Provide the micro-nano fluidic sorting system according to claim 5, and prepare the biological particles to be sorted into a to-be-sorted liquid; S22) Input the to-be-sorted liquid into the sample channel of the high-throughput micro-nano fluidic control device through the third input module, and through the cascade connection between the third flow control module and the third circulation module, enable the to-be-sorted liquid to circulate in the sample channel; the biological particles are sorted according to their particle sizes in the compression channel, and the biological particles passing through the compression channel finally enter the microchannel array; S23) Collect the biological particles in the microchannel array and the biological particles in the sample channel; Preferably, in S21), the high-throughput micro-nano fluidic control device includes at least two groups of microchannel arrays, and the diameters of the compression channels connecting the sample channel to different microchannel arrays are different; Preferably, in S23), it further includes the step of further concentrating and purifying the sorted biological particles.

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