Exosome enrichment micro-fluidic chip based on induced charge electroosmosis micro-vortex

Abstract

The utility model relates to the technical field of exosome enrichment, and discloses an exosome enrichment micro-fluidic chip based on induced charge electroosmosis micro vortexes, which comprises a bottom plate and a cover plate, a first excitation electrode and a second excitation electrode are oppositely arranged on the bottom plate, and a trapezoidal suspension electrode is arranged between the first excitation electrode and the second excitation electrode; an oil phase inlet, a water phase inlet, a liquid drop channel and a plurality of outlets are formed in the cover plate, and the trapezoidal suspension electrode is located in the liquid drop channel; the plurality of outlets are communicated with one end of the liquid drop channel through the outflow channel; mixed particles are introduced from a water phase inlet and are sheared by an oil phase to form liquid drops, the liquid drops flow into a liquid drop channel, and the morphology of an induced charge electroosmosis micro vortex can be regulated and controlled by regulating and controlling the voltage and frequency of a trapezoidal suspension electrode, so that the exosome is enriched below the channel; the liquid drop is split into a plurality of sub-liquid drops at the tail part of the liquid drop channel, and one of the sub-liquid drops is the sub-liquid drop after particle concentration, so that the enrichment effect of the exosome is realized.

Description

TECHNICAL FIELD

[0001] The utility model relates to exosome enrichment technical field, concretely relates to a kind of exosome enrichment microfluidic chip based on induced charge electroosmosis micro-vortex. BACKGROUND

[0002] Exosomes are nanovesicles of about 30-150 nm in diameter secreted by cells, which play a key role in intercellular information transmission and can reflect the health status and function of cells. Therefore, exosomes have great potential in disease diagnosis and treatment. To fully utilize this potential of exosomes, capture and analysis techniques are essential. Efficient capture methods can improve the purity and collection efficiency of exosomes, thereby enhancing the accuracy and reliability of subsequent analysis. However, due to the small size of exosomes and their low content in body fluids, precise capture and separation of exosomes remains a challenging task.

[0003] Currently, the main methods for capturing exosomes include ultracentrifugation, density gradient centrifugation, and ultrafiltration. Among them, ultracentrifugation is the most common method, which separates exosomes by the difference in sedimentation velocity of different particles in solution. However, this method has several obvious defects: (1) time-consuming operation, low efficiency; (2) large fluctuation in recovery rate, unsatisfactory separation purity; (3) high equipment cost. Density gradient centrifugation separates exosomes by layering cell components using different density media. Although this method can effectively separate exosomes, it is complex, time-consuming, and yields a small amount of exosomes. In addition, the high permeability of gradient media may cause exosome dehydration, thereby affecting its biological function. Ultrafiltration separates exosomes by selecting a filter membrane with appropriate pore size based on the size of exosomes. This method is simple, low-cost, and efficient, but impurities smaller than the filter membrane pore size or flexible proteins may also pass through the filter membrane, reducing the purity of exosomes. Meanwhile, membrane pore blockage may also affect separation efficiency.

[0004] Therefore, the utility model provides an exosome capture microfluidic chip based on induced charge electroosmosis and end multi-outlet structure. UTILITY MODEL CONTENT

[0005] The utility model aims at overcoming the deficiencies in the prior art and providing an exosome enrichment microfluidic chip based on induced charge electroosmosis micro-vortex. By applying an electric field in the channel, the chip induces exosomes to flow out from the end outlet at the lowest position based on the principle of induced charge electroosmosis, achieving the enrichment effect of exosomes.

[0006] To achieve the above-mentioned purpose, the utility model adopts the following technical solutions:

[0007] A microfluidic chip for enriching exosomes based on induced charge electroosmotic microvortices comprises a base plate and a cover plate mounted above the base plate, wherein a first excitation electrode and a second excitation electrode are arranged opposite each other on the base plate, and a trapezoidal suspended electrode is arranged between the two; an oil phase inlet, a water phase inlet, a droplet channel, and multiple outlets are formed on the cover plate, and the trapezoidal suspended electrode is located in the droplet channel; the oil phase inlet is connected to the droplet channel through the oil phase channel, the water phase inlet is connected to the droplet channel through the water phase channel, and the water phase channel and the oil phase channel intersect at one end of the droplet channel, and the multiple outlets are all connected to the other end of the droplet channel in a divergent manner through independent outflow channels.

[0008] Optionally, the oil phase channel and the water phase channel are both opened at the bottom of the cover plate, the oil phase channel is a rectangular structure, and the water phase inlet and the water phase channel are both located on the inner side of the rectangular structure.

[0009] Optionally, there are four outlets, namely a first outlet, a second outlet, a third outlet and a fourth outlet.

[0010] Optionally, the first outlet, the second outlet, the third outlet and the fourth outlet are respectively connected to collection pipes for discharging liquid.

[0011] Optionally, the oil phase inlet and the water phase inlet are respectively connected to hoses for injecting liquid.

[0012] Optionally, the long side of the trapezoidal suspension electrode is close to the water phase inlet, and the short side of the trapezoidal suspension electrode is close to the outlet.

[0013] Optionally, the hypotenuse of the trapezoidal floating electrode is close to the first exciting electrode.

[0014] Optionally, the output end of the signal generator is connected to the input end of the signal amplifier, the positive electrode of the output end of the signal amplifier is connected to the first excitation electrode, and the negative electrode is connected to the second excitation electrode.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0016] (1) First, the mixed particles are introduced from the water phase inlet, form droplets after shearing in the oil phase, and then flow into the droplet channel. By adjusting the voltage and frequency of the trapezoidal suspended electrode in the droplet channel, the morphology of the induced charge electroosmotic micro-vortex can be controlled, and then the exosomes are enriched under the channel. The droplets are split into multiple sub-droplets at the tail of the droplet channel through a multi-branch structure, one of which is a sub-droplet after particle concentration, and the rest are sub-droplets without particles, thereby realizing the enrichment effect of exosomes; the separation and enrichment steps of the chip are simple, time-consuming and efficient, and the separation purity is ideal, and a large number of exosomes are obtained;

[0017] (2) Compared with the traditional capture method, the utility model directly enriches exosomes through a micro-channel structure by a physical method, reduces the loss caused by multiple treatments and transfers, and improves the efficiency. At the same time, compared with the traditional ultracentrifugation method and density gradient centrifugation method, the method needs complex pretreatment steps, and the operation is more simple, which reduces the experimental time and cost. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 is a structural schematic diagram of an exosome enrichment microfluidic chip according to an embodiment of the utility model;

[0019] Figure 2 is a structural schematic diagram of an exosome enrichment microfluidic chip according to an embodiment of the utility model;

[0020] Figure 3 is Figure 2 is a local enlarged schematic view of A in FIG. 1;

[0021] Figure 4 is a structural schematic diagram of the bottom plate according to an embodiment of the utility model;

[0022] Figure 5 is a structural schematic diagram of the cover plate according to an embodiment of the utility model;

[0023] Figure 6 is a structural schematic diagram of the bottom plate according to an embodiment of the utility model;

[0024] Figure 7 is a structural schematic diagram of the bottom plate according to an embodiment of the utility model;

[0025] 1, bottom plate; 101, first excitation electrode; 102, second excitation electrode; 2, cover plate; 3, trapezoidal suspended electrode; 4, oil phase inlet; 401, oil phase channel; 5, water phase inlet; 501, water phase channel; 601, first outlet; 602, second outlet; 603, third outlet; 604, fourth outlet; 7, outflow channel; 8, droplet channel. DETAILED DESCRIPTION

[0026] The utility model will be further explained in detail in connection with the drawings and embodiments, these drawings are all simplified schematic diagram, only with the schematic way the basic structure of the utility model is explained, therefore it only shows the related structure of the utility model.

[0027] Embodiment one

[0028] As Figures 1-7 Induced charge electroosmotic micro-vortex based on exosome enrichment microfluidic chip, including bottom plate 1 and cover plate 2, bottom plate 1 adopts glass sheet, cover plate 2 adopts PDMS material to be made, PDMS is polydimethylsiloxane, it is a widely used organic silicon polymer, its processing is relatively easy, can be mixed after with crosslinking agent pours into mould, under certain temperature heating makes it solidify, thereby obtains the product of required shape, this processing mode is simple, low in cost, and can realize the structure forming of high precision, can even shape several nanometer structures.

[0029] Cover plate 2 is arranged above bottom plate 1, and first excitation electrode 101, second excitation electrode 102 and trapezoidal suspension electrode 3 are arranged on bottom plate 1, and first excitation electrode 101 and second excitation electrode 102 are oppositely arranged on both sides of trapezoidal suspension electrode 3.

[0030] Oil phase inlet 4, water phase inlet 5, droplet channel 8 and multiple outlets are formed on cover plate 2, trapezoidal suspension electrode 3 is located in droplet channel 8, oil phase inlet 4 is communicated with droplet channel 8 through oil phase channel 401, water phase inlet 5 is communicated with droplet channel 8 through water phase channel 501, and water phase channel 501 and oil phase channel 401 meet at one end of droplet channel 8, and multiple outlets are communicated with the other end of droplet channel 8 in a divergent manner through independent outflow channels 7.

[0031] Concentration principle: at the inlet of droplet channel 8, the oil phase can wrap the solution in the continuous fluid into discrete microdroplets by the principle of fluid shear; voltage is applied on first excitation electrode 101 and second excitation electrode 102, so that an electric field is generated in droplet channel 8, the electric field can induce the migration of free electrons on trapezoidal suspension electrode 3, and finally a positive and negative double layer is formed on trapezoidal suspension electrode 3, the double layer moves in the opposite direction along the electric field line to form an induced charge electroosmotic (ICEO) vortex, and then the exosomes are concentrated in the ICEO vortex.

[0032] Firstly, the mixed particles are introduced from the water phase inlet 5, and after being sheared by the oil phase to form droplets, the droplets flow into the droplet channel 8. By adjusting the voltage and frequency of the trapezoidal suspension electrode 3 in the droplet channel 8, the morphology of the induced charge electroosmotic micro-vortex can be controlled, and then the exosomes are enriched under the droplet channel 8. The droplets are split into multiple sub-droplets at the tail of the droplet channel 8 through a multi-branch structure, one of which is a sub-droplet after particle concentration, and the rest are sub-droplets without particles, thereby realizing the enrichment effect of exosomes.

[0033] The chip is used to separate and enrich exosomes, which has the advantages of simple separation and enrichment steps, short time consumption, high efficiency, ideal separation purity, and large number of obtained exosomes.

[0034] The oil phase channel 401 and the water phase channel 501 are both arranged at the bottom of the cover plate 2, the oil phase channel 401 has a rectangular structure, and the water phase inlet 5 and the water phase channel 501 are both located inside the rectangular structure. The mixed particles are introduced from the water phase inlet 5 and flow along the water phase channel 501 to the intersection, and at this time the oil phase is introduced from the oil phase inlet 4. By using the principle of fluid shear, the continuous mixed particle solution is wrapped into discrete small droplets along the oil phase channels 401 on both sides.

[0035] Further, the outlet is provided with four first outlets 601, second outlets 602, third outlets 603 and fourth outlets 604 from top to bottom. Since the multiple outflow channels 7 are divergently connected to the end of the droplet channel 8, a multi-branch structure is formed, which can split the droplets into four sub-droplets, one of which is a sub-droplet after particle concentration, and the rest are sub-droplets without particles. The sub-droplet after particle concentration is close to the outflow channel 7 corresponding to the lowermost fourth outlet 604.

[0036] The long side of the trapezoidal suspension electrode 3 is close to the water phase inlet 5, the short side of the trapezoidal suspension electrode 3 is close to the outlet, and the oblique side of the trapezoidal suspension electrode 3 is close to the first excitation electrode 101. The trapezoid has a group of parallel sides, and the shorter parallel side can be regarded as the "short side" and the longer parallel side can be regarded as the "long side". In this way, the effect of directional transport of particles can be achieved, and it is ensured that the sub-droplet after particle concentration can enter the outflow channel 7 corresponding to the fourth outlet 604.

[0037] Directional transport: the trapezoidal suspended electrode 3 in the droplet channel 8 is an equipotential body, due to the asymmetry of the trapezoidal structure, as the droplet channel 8 goes deeper, the potential difference between the second excitation electrode 102 and the trapezoidal suspended electrode 3 remains unchanged, while the potential difference between the first excitation electrode 101 and the trapezoidal suspended electrode 3 decreases, so that the relative difference of the potential difference on both sides increases, and the ICEO vortex formed near the second excitation electrode 102 side is stronger. The asymmetric vortex in the droplet channel 8 will introduce nanoparticles into the vortex with higher intensity, and then into the fourth outlet 604 with the droplet, realizing the effect of directional transport of particles.

[0038] At the end of the droplet channel 8, the droplet will be split into four small droplets through the multi-branch structure. In order to effectively control the uniformity of the droplet in the splitting process and ensure that the sizes of the droplets in the four outflow channels 7 are as consistent as possible, an equal amount of pure water is injected into each outflow channel 7 before the experiment; by adjusting the liquid level in the outlet, the pressure in the four outflow channels 7 is ensured to be equal, thereby maintaining the consistency of the pressure in each channel.

[0039] In addition, the diameter of the water phase inlet 5 and the channel width is 100 microns, the diameter of the oil phase inlet 4 is 100 microns, the width of the oil phase channel 401 is 100 microns, the width of the droplet channel 8 is 100 microns, the long side width of the trapezoidal suspended electrode 3 is 95 microns, the short side width is 10 microns, and the width of the four branched outlets connected with the channels is 25 microns, and the diameter of the four outlets is 100 microns, that is, the diameter of the axial outflow channel 7 gradually increases to the outlet position.

[0040] Example two

[0041] Based on the embodiment one, the utility model discloses a kind of preparation methods of exosome enrichment microfluidic chip.

[0042] Glass sheet with ITO film and photoresist AZ4620 are used to process electrode structure based on standard soft lithography technology. Bottom plate 1 uses glass sheet with ITO film, and electrode structure is processed on glass sheet based on standard soft lithography technology using photoresist. The electrode structure includes first excitation electrode 101, second excitation electrode 102 and trapezoidal suspended electrode 3. First excitation electrode 101 and second excitation electrode 102 are oppositely distributed on both sides of trapezoidal suspended electrode 3.

[0043] Channel mold is processed using Dupont dry film based on standard soft lithography technology. The channel mold has oil phase channel 401, water phase channel 501, outflow channel 7 and droplet channel 8. Then, PDMS melt is poured into the channel mold to process cover plate 2 with channel structure.

[0044] A 1mm hole is punched at the inlet location using a puncher, and four 100-micron diameter holes are punched at the outlet locations.

[0045] An oil phase inlet 4 and a water phase inlet 5 are respectively formed at the ends of the oil phase channel 401 and the water phase channel 501 using a puncher, and a plurality of outlets are formed at the end of the outflow channel 7; the diameters of the oil phase inlet 4 and the water phase inlet 5 are 1mm, and the diameter of the outlet is 100 microns.

[0046] After the cover plate 2 and the bottom plate 1 are subjected to hydrophilic treatment, they are bonded together; hydrophilic treatment is a surface treatment technology aimed at changing the surface of a material from a hydrophobic (water-repellent) state to a hydrophilic (water-attracting) state. This treatment can change the physical and chemical properties of the material's surface, increasing its affinity for water molecules. Bonding refers to the process of tightly connecting two or more materials together through physical or chemical methods. This connection allows the materials to form a stable bond, meeting specific functional requirements such as increased mechanical strength, improved electrical performance, or enhanced sealing.

[0047] A 100-micron outer diameter rubber tube is inserted into the oil phase inlet 4 and the water phase inlet 5, and AB glue (a two-component adhesive) is used for bonding, to facilitate the injection of samples through a micro pump during the experiment; the first outlet 601, the second outlet 602, the third outlet 603, and the fourth outlet 604 are respectively connected to collection tubes for discharging liquids; the micro pump needs to be continuously injected during the experiment, and the injected liquid will eventually be discharged through the four outlet-connected chip external collection tubes and other collection channels, collected by a collection device such as a culture dish, and then transferred.

[0048] In addition, since the droplets generated in this experiment are water-in-oil droplets, the continuous phase is the oil phase and the dispersed phase is the water phase, so the microchannel needs to be hydrophobic before the experiment. In order to generate stable water-in-oil single emulsion droplets, Aquapel glass anti-fog agent is injected into the channel structure, soaked for 2 minutes, then the channel is washed with acetone, and placed in a constant temperature drying oven at 80℃ for 30 minutes to dry, thereby obtaining a hydrophobic channel surface; the channel structure here is the oil phase channel 401, the water phase channel 501, the outflow channel 7, and the droplet channel 8.

[0049] Example Three

[0050] Based on the embodiment one, the utility model discloses a method for separating and enriching exosomes, which uses the above-mentioned exosome enrichment microfluidic chip based on induced charge electroosmosis microvortex.

[0051] The output end of the signal generator is connected with the input end of the signal amplifier, the positive electrode of the output end of the signal amplifier is connected with the first excitation electrode 101, and the negative electrode is connected with the second excitation electrode 102; one end of the rubber tube is respectively inserted into the oil phase inlet 4 and the water phase inlet 5, and the other end is connected to the output end of the injection micro pump by using AB glue; meanwhile, one end of the collection tube is respectively inserted into the first outlet 601, the second outlet 602, the third outlet 603 and the fourth outlet 604, and the other end is respectively placed in different culture dishes to collect the discharged liquid and exosomes.

[0052] The experimental process is as follows:

[0053] (1) A certain amount of pure water is injected into each outflow channel 7 to adjust the liquid level of the outlet and ensure that the pressure at the outlet is consistent;

[0054] (2) Check the connection between the chip electrode and the output end of the signal amplifier, and turn on the signal generator;

[0055] (3) The micro flow injection pump is used to inject the silicon oil-PDMS (mass ratio 1:1) oil phase from the oil phase inlet 4, and after the input is stable, the exosome glycerol mixed solution is introduced into the water phase inlet 5; adjust the flow rates of the two according to the actual situation until the droplets are generated stably at the water-oil phase junction by using the fluid shear principle;

[0056] (4) At the outflow channel 7, four collection tubes are used to collect the droplets into culture dishes, and the fourth outlet 604 corresponds to the output of the enriched exosome droplets.

[0057] Among them, the shear-generated droplets contain a low concentration of exosome and glycerol mixed solution, and the droplets pass through the enrichment principle in the droplet channel 8, so that the exosomes are gathered in a specific area of the droplet (roughly parallel to the channel corresponding to the fourth outlet 604). At the outflow channel 7, the droplets are divided into four small droplets by the bifurcation structure; due to the distribution characteristics of the exosomes inside the original droplet, the exosome concentration in the first three small droplets is low, while the exosome concentration in the last small droplet is the highest, thereby realizing the separation and enrichment of the exosomes. The four channels further reduce the size of the original channel, making the enrichment area narrower, so that higher concentration of exosomes can be collected.

[0058] Specifically, the sample is injected into the chip by the injection micro pump, and during the experiment, the output end of the signal generator is connected with the input end of the signal amplifier, the positive electrode of the output end of the signal amplifier is connected with the first excitation electrode 101, and the negative electrode is connected with the second excitation electrode 102. After power-on, the performance of the sample in the chip is observed, the input flow rate of the injection pump, the frequency and amplitude of the voltage are adjusted to find the suitable separation conditions, so that the exosomes enter the droplets in the outflow channel 7 corresponding to the fourth outlet 604.

[0059] Before the micro-mixing experiment, a mixture of glycerol and deionized water with a mass ratio of 0%, 20%, and 50% is prepared, and then potassium chloride is added to the mixture to prepare a buffer with a conductivity of 0.1 S / m, 0.2 S / m, and 0.5 S / m, respectively. In the micro-reaction experiment, ferric chloride and potassium ferrocyanide are added to the glycerol mixture with a mass ratio of 20%, and the conductivity is calibrated to 0.2 S / m by using a conductivity meter.

[0060] Here, three different mixtures are used, which differ in glycerol concentration. The higher the mass ratio, the larger the final droplet size, which can be used to control the droplet size. The conductivity is 0.2.

[0061] The water phase inlet 5 is connected to the glycerol mixture containing exosomes, and the final result is the enriched exosome droplets; the water phase refers to the exosome glycerol mixture (the exosome culture solution is added to the glycerol and deionized water mixture with different mass ratios described above). The oil phase is a PDMS silicone oil mixture with a mass ratio of 1:1. The exosomes are added to the glycerol and deionized water mixture, and the water phase is injected. The potassium chloride buffer is used to adjust the conductivity to 0.2 S / m, which is the solution conductivity condition for realizing enrichment.

[0062] In order to effectively control the uniformity of the droplets during the splitting process and ensure that the droplet sizes in the four channels are as consistent as possible, an equal amount of pure water is injected into each outflow channel 7 before the experiment, and the height of the liquid level in the outlet is adjusted to ensure that the pressures in the various outflow channels 7 are equal, thereby maintaining the consistency of the pressure and ensuring that the sizes of the split droplets are consistent.

[0063] In summary, the microfluidic chip mainly includes two layers: the lower layer is a glass plate with a planar electrode, and the upper layer is a PDMS material containing a microchannel structure, which is the core of the entire chip structure. The end has four outflow channels 7 for exosome enrichment and purification. By applying an electric field in the droplet channel 8, based on a series of principles such as induced charge electroosmosis, exosomes are induced to flow out from the lowest end outlet, and the enrichment effect of exosomes can be achieved. Compared with the traditional capture method, the utility model directly enriches exosomes through a microchannel structure by a physical method, reduces the loss caused by multiple processing and transfer, and improves the efficiency; at the same time, compared with the traditional ultracentrifugation method and density gradient centrifugation method which require complex pretreatment steps, this method is more simple to operate, and reduces the experimental time and cost.

[0064] In the description of the utility model, it is necessary to understand that the orientation or positional relation indicated by the terms "center", "longitudinal", "transverse", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer" and the like is the orientation or positional relation based on the orientation or positional relation shown in the drawings, which is only for the convenience of describing the utility model and simplifying the description, and does not indicate or imply that the device or element indicated must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the utility model. In addition, the terms "first", "second" and the like are only for the purpose of description, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the technical features indicated. Therefore, the features limited by "first", "second" and the like can explicitly or implicitly include one or more features. In the description of the utility model, unless otherwise specified, the meaning of "a plurality of" is two or more.

[0065] In the description of the utility model, it should be explained that, unless otherwise specified and limited, the terms "mounting", "connection", "connection" should be understood broadly, for example, it can be fixed connection, can also be detachable connection, or integrally connected, can be mechanical connection, can also be electrical connection, can be directly connected, can also be indirectly connected through an intermediate medium, can be the communication inside two elements. For ordinary skilled in the art, the specific meaning of the above terms in the utility model can be understood through specific circumstances.

[0066] According to the ideal embodiments of the utility model, the above description, relevant personnel can make various changes and modifications without deviating from the technical idea of the utility model. The technical scope of the utility model is not limited to the contents in the specification, and the technical scope must be determined according to the scope of claims.

Claims

1. A microfluidic chip for enriching exosomes based on induced charge electroosmotic microvortices, characterized by: It comprises a base plate (1) and a cover plate (2) mounted above the base plate (1); a first excitation electrode (101) and a second excitation electrode (102) are arranged on the base plate (1) in a relative manner, and a trapezoidal suspension electrode (3) is arranged between the two; The cover plate (2) is provided with an oil phase inlet (4), a water phase inlet (5), a liquid droplet channel (8) and a plurality of outlets, and the trapezoidal suspension electrode (3) is located in the liquid droplet channel (8); The oil phase inlet (4) is connected to the droplet channel (8) through the oil phase channel (401), and the water phase inlet (5) is connected to the droplet channel (8) through the water phase channel (501), and the water phase channel (501) and the oil phase channel (401) intersect at one end of the droplet channel (8), and the multiple outlets are all connected to the other end of the droplet channel (8) in a divergent manner through independent outflow channels (7).

2. The exosome enrichment microfluidic chip based on induced charge electroosmotic micro-vortex according to claim 1, characterized in that: The oil phase channel (401) and the water phase channel (501) are both opened at the bottom of the cover plate (2), the oil phase channel (401) is a rectangular structure, and the water phase inlet (5) and the water phase channel (501) are both located on the inner side of the rectangular structure.

3. The exosome enrichment microfluidic chip based on induced charge electroosmotic micro-vortex according to claim 1, characterized in that: There are four outlets, namely a first outlet (601), a second outlet (602), a third outlet (603) and a fourth outlet (604).

4. The exosome enrichment microfluidic chip based on induced charge electroosmotic micro-vortices according to claim 3, characterized in that: The first outlet (601), the second outlet (602), the third outlet (603) and the fourth outlet (604) are respectively connected to a collecting pipe for discharging liquid.

5. The exosome enrichment microfluidic chip based on induced charge electroosmotic micro-vortices according to claim 4, characterized in that: The oil phase inlet (4) and the water phase inlet (5) are respectively connected to hoses for injecting liquid.

6. The exosome enrichment microfluidic chip based on induced charge electroosmotic micro-vortex according to claim 1, characterized in that: The long side of the trapezoidal suspension electrode (3) is close to the water phase inlet (5), and the short side of the trapezoidal suspension electrode (3) is close to the outlet.

7. The exosome enrichment microfluidic chip based on induced charge electroosmotic micro-vortices according to claim 6, characterized in that: The oblique side of the trapezoidal floating electrode (3) is close to the first excitation electrode (101).

8. The exosome enrichment microfluidic chip based on induced charge electroosmotic micro-vortices according to claim 7, characterized in that: The output end of the signal generator is connected to the input end of the signal amplifier, the positive electrode of the output end of the signal amplifier is connected to the first excitation electrode (101), and the negative electrode is connected to the second excitation electrode (102).

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