A plasma exosome filtration and extraction device and its usage method

Abstract

The present invention discloses a plasma exosome filtration and extraction device and its usage method. The filtration and extraction device includes: a cryogenic centrifuge, a dilution dispenser, a shaker, a filter, and an exosome collection box. The cryogenic centrifuge is arranged at the front end of the filtration and extraction device and is used for performing cryogenic centrifugation on the pre-treated samples after thawing and balancing. A dilution dispenser is arranged on one side of the cryogenic centrifuge. The dilution dispenser includes a distribution chamber, a shaker arranged at the bottom side of the distribution chamber, a dilution tube arranged in the distribution chamber, an input pump head, an output pump head, and an input suction tube and an output suction tube respectively connected to the input pump head and the output pump head. The end of the input suction tube is branched into a plurality of input heads, and the plurality of input heads can extend into the upper layer of the sample tube after the cryogenic centrifuge completes the centrifugation of the sample to collect the supernatant into the distribution chamber. The present invention can process exosome samples in batches and automatically, and further improve the efficiency of separating exosomes in the laboratory.

Description

Technical Field

[0001] The present invention relates to the field of plasma exosome extraction, and specifically to a plasma exosome filtration extraction device and its usage method. Background Art

[0002] Exosomes are small vesicles with a diameter of about 30 - 150 nm secreted by living cells, having a typical lipid bilayer structure, and existing in cell culture supernatants, serum, plasma, saliva, urine, amniotic fluid, and other biological fluids. Exosomes carry a variety of important information such as proteins, lipids, and RNAs. They not only play an important role in the material and information transfer between cells, but also are expected to become early diagnostic markers for various diseases. Tumors occur before tiny lesions can be detected by imaging. By using tumor cells, cfDNA, or exosomes in the blood and detecting the content of markers directly related to tumors such as cfDNA and CTC in the blood through means such as PCR and high-throughput sequencing, early tumors that cannot be identified by imaging can be discovered. Detecting tumors early and intervening can reduce the incidence and mortality of tumors.

[0003] The extraction of exosomes in the prior art generally includes the following process:

[0004] Step 1: Collect a peripheral blood sample from a volunteer using an anticoagulant tube containing ethylenediaminetetraacetic acid and store it at room temperature for 10 min;

[0005] Step 2: After sample balancing, centrifuge at 500 x g for 10 min to remove formed elements such as cells and fibers in the blood. Then use a pipette tip to transfer the supernatant to a 2 mL sterile and enzyme-free EP tube;

[0006] Step 3: After EP tube balancing, set a low-temperature centrifuge at 4 °C and centrifuge at 3000 x g for 20 min to remove apoptotic bodies and cell debris, etc. Then, use a pipette tip to aspirate the supernatant and transfer it to a 2 mL sterile and enzyme-free EP tube;

[0007] Step 4: After EP tube balancing, set a low-temperature centrifuge at 4 °C and centrifuge at 12000 x g for 20 min to remove large vesicles. Then, use a pipette tip to collect the supernatant and transfer it to a 2 mL sterile and enzyme-free EP tube. Freeze it at -80 °C for 24 h;

[0008] Step 5: Take out the frozen sample, place it on ice to thaw. After the sample is completely thawed, balance the EP tube, and then obtain exosomes after filtration extraction and ultracentrifugation.

[0009] For the above-mentioned exosome extraction method, for example, there is also a method for separating and extracting exosomes disclosed in the publication number CN117721062A. The general steps include: Step 1: Mix the biological sample to be separated with PBS buffer in a certain proportion, and use a stirrer to stir it at a stirring speed of 60-180 rpm. In the present invention, by mixing the biological sample with PBS buffer, the structure and biological activity of exosomes can be better protected. By separating the precipitate from the supernatant, the supernatant is filtered through a dynamic filter membrane device, and the supernatant obtained after centrifugation of the washed filter membrane is collected. This is repeated at least twice to ensure the separation yield. The collected supernatant is centrifuged with an ultracentrifuge, and exosomes are separated and collected through a dynamic filter membrane device with a smaller pore size of the filter membrane, thereby improving the separation effect and purity; by using technical means such as stirring with a stirrer and centrifuging with a centrifuge, the exosomes in the biological sample can be effectively and quickly separated and extracted, so as to improve the separation and extraction efficiency;

[0010] Both of the above two processes point out that the main measures for exosome extraction are: filtration extraction, which generally includes processes such as low-temperature centrifugation, PBS shaking dilution, and filter filtration. Since this process is relatively cumbersome, manual operation is currently used in the existing technology. Manual operation has low efficiency when dealing with batch samples. Therefore, a plasma exosome filtration extraction device and its use method are needed to handle this filtration extraction process, which can batch and automate the treatment of exosome samples after steps 1-4, and further improve the efficiency of separating exosomes in the laboratory. Summary of the Invention

[0011] The purpose of the present invention is to provide a plasma exosome filtration extraction device and its use method, which can batch and automate the treatment of exosome samples through this device and its use method, and further improve the efficiency of separating exosomes in the laboratory.

[0012] To achieve the above purpose, the present invention provides the following technical solutions: A plasma exosome filtration extraction device, including:

[0013] A low-temperature centrifuge, configured at the front end of the filtration extraction device and used for low-temperature centrifugation of the pre-treated sample after thawing and balancing. A dilution dispenser is provided on one side of the low-temperature centrifuge;

[0014] The dilution dispenser includes a distribution chamber, an oscillator provided at the bottom side of the distribution chamber, a dilution tube provided in the distribution chamber, an input pump head, an output pump head, and an input suction tube and an output suction tube respectively connected to the input pump head and the output pump head;

[0015] A shaker, which is used to shake and mix the diluted sample. The shaker includes a frame, an upper frame, a swing-type shaking component, a shaking tray and a shaking tube module. The upper frame is arranged along the end face of the frame. One-way sliding frames are arranged on both sides of the upper frame, and the shaking tray can be connected in a guiding manner inside the upper frame;

[0016] A filter, which is used to filter the sample after dilution, shaking and homogenization. The filter includes a syringe-type pre-filter, a filter tube and a pushing module;

[0017] An exosome collection box, which can be connected to the output end of the filter tube through multiple connectors.

[0018] Preferably, the end of the input suction tube is branched into multiple input heads, and the multiple input heads can extend into the upper layer of the sample tube after the sample is centrifuged in a low-temperature centrifuge to collect the supernatant into the distribution bin; the dilution tube can be connected to an external PBS diluent, and the output side of the output suction tube is connected to the shaker.

[0019] Preferably, the output suction tube is connected to a distribution rack through the end of a hose. The bottom side of the distribution rack is branched into multiple distribution output heads, and the number of the distribution output heads matches the number of the shaking tube modules on the shaking tray. The distribution output heads can extend into the shaking tube modules.

[0020] Preferably, the swing-type shaking component includes a shaking arm, a swing plate, a disc, a motor A, a large pulley and a small pulley. Two groups of discs are provided and are coaxially rotatably connected to both sides of the upper frame. A large pulley is arranged at the rear end of one of the discs. The large pulley is linked with the small pulley through a belt, and the small pulley is connected to the shaft end of the motor A; a slide plate is arranged on the end face of the large pulley, a swing plate is arranged in front of the large pulley, a one-way slot for sliding connection with the slide plate is arranged on the swing plate, the lower end of the swing plate is hinged to the frame, and the upper end of the swing plate is hinged to the shaking arm.

[0021] Preferably, a slide shaft is arranged inside the one-way sliding frame, and the end of the slide shaft can be connected to the shaking tray. The end of the shaking arm is hinged to the slide shaft, and the shaking arm can drive the shaking tray to move back and forth when the turntable rotates; a plurality of slot holes are arranged in an array on the shaking tray, and each slot hole is provided with a shaking tube module. The shaking tube module includes a tube body, an upper cover plate and a confluence tube. The upper cover plate is arranged at the upper end of the tube body, an outlet tube is connected to the bottom side of the tube body, an electromagnetic check valve is arranged on the outlet tube, the ends of the outlet tubes of the multiple shaking tube modules are connected to the confluence tube, the confluence tube is a hose, and the end of the confluence tube is branched into three pipelines to be connected to the filter, and a check valve is arranged on each pipeline.

[0022] Preferably, at least three groups of syringe-type pre-filters are provided. The three groups of syringe-type pre-filters are located on one side of the pushing module and are fixed to the bottom side through a bracket. The syringe-type pre-filter includes a syringe body, a piston push plate and a pre-filter layer. The pre-filter layer is arranged on the front side of the syringe body, the piston push plate is arranged inside the syringe body, and the rear end of the piston push plate extends out of the syringe body and is close to the pushing module.

[0023] Preferably, the pushing module includes a T-shaped bracket, a track, a rack push plate, a gear set and a driving unit. A track is provided on the bottom side of the T-shaped bracket, and the driving unit is installed on the T-shaped bracket. The rack push plate can be slidably connected to the track, and the front end of the rack push plate is close to the piston push plate. The driving unit includes a motor B installed on the upper side of the T-shaped bracket and a gear set linked by a belt and a pulley. The gear set includes three gears connected coaxially. The three gears are respectively engaged with the rack push plate and push the rack push plate to slide along the track to push the piston push plate. The filter tube is connected to the front end of the syringe body through a joint tube, and multiple filter layers for filtering exosome samples are provided in the filter tube.

[0024] The present invention also provides a method for using the above-mentioned plasma exosome filtration and extraction device, and the specific steps include:

[0025] S1: Take out the frozen sample, place it on ice for thawing. After the sample is completely thawed, balance the EP tube, and then place the sample in a low-temperature centrifuge for low-temperature centrifugation to remove large vesicles;

[0026] S2: Use the input pump head and input suction tube of the dilution dispenser to suck and collect the sample supernatant into the distribution bin. The distribution bin is diluted by adding PBS diluent through a dilution tube, then pre-shake using a shaker, and then use the output suction tube to divert the diluted sample into multiple oscillation tube modules of the shaker;

[0027] S3: Multiple oscillation tube modules can achieve the oscillation effect by moving back and forth using an oscillation tray. After multiple reciprocating oscillations, the electromagnetic one-way valve at the bottom of the tube body is opened, and the samples in multiple tube bodies flow together through a confluence tube into the syringe pre-filter;

[0028] S4: The syringe pre-filter can use the gear to engage with the rack push plate and the piston push plate. The piston push plate can filter and push out the sample entering the syringe body, so that the sample enters the filter tube at high speed for filtration to remove impurities. The filtered sample is collected through an exosome collection box and completed exosome filtration and extraction after ultracentrifugation.

[0029] Compared with the prior art, the beneficial effects of the present invention are:

[0030] 1. The present invention can process exosome samples in batches and automatically, further improving the efficiency of separating exosomes in the laboratory.

[0031] 2. The present invention configures a dilution dispenser between a low-temperature centrifuge and a shaker. This type of dilution dispenser can add PBS diluent for dilution, then use the shaker for pre-shaking, and then use the output suction tube to divert the diluted sample into multiple oscillation tube modules of the shaker. The shaker performs an oscillation operation on the oscillation tube modules. The advantage of the dilution dispenser is that it can quickly transfer a large number of samples in the low-temperature centrifuge and achieve sample dilution through pre-shaking and introducing PBS.

[0032] 3. The shaker of the present invention is used to shake the diluted sample to homogenize the sample, and the whole shaking and homogenizing process is convenient and fast.

[0033] 4. The filter of the present invention is based on the combination of a syringe-type pre-filter and a filter tube. The syringe-type pre-filter can be filled with a filter layer in a disposable syringe and automatically achieve automatic filtration by pushing through a pushing module. The sample enters the filter tube for deep filtration after passing through the syringe-type pre-filter. Since the pushing module can pressurize the disposable syringe, the sample after entering the filter tube has pressure and can achieve power-free filtration, and the impurity filtration speed is fast. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 is a schematic diagram of the overall structure of the present invention;

[0035] Figure 2 is a schematic diagram of the structure of the low-temperature centrifuge and the dilution dispenser in the embodiment of the present invention;

[0036] Figure 3 is a schematic diagram of the structure of the shaker in the embodiment of the present invention;

[0037] Figure 4 is a schematic diagram of the structure of the oscillation tube module in the embodiment of the present invention;

[0038] Figure 5 is a schematic diagram of the structure of the filter and the exosome collection box in the embodiment of the present invention.

[0039] In the figure: 1. Low-temperature centrifuge; 2. Dilution dispenser; 201. Distribution bin; 202. Dilution tube; 203. Output suction tube; 204. Distribution rack; 205. Distribution output head; 3. Shaker; 301. Frame; 302. Upper frame; 303. Linear slide; 304. Oscillation arm; 305. Swing plate; 306. Disc; 307. Large pulley; 308. Small pulley; 309. Slide plate; 310. Oscillation tray; 311. Oscillation tube module; 311a. Tube body; 311b. Upper cover plate; 311c. Confluence tube; 311d. Electromagnetic check valve; 4. Filter; 401. Syringe body; 402. Piston push plate; 403. T-shaped bracket; 404. Track; 405. Rack push plate; 406. Gear set; 407. Filter tube; 5. Exosome collection box. Detailed implementation mode

[0040] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0041] In the description of the present invention, it should be noted that the orientation or positional relationship indicated by the terms "vertical", "upper", "lower", "horizontal", etc. is based on the orientation or positional relationship shown in the accompanying drawings. It is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be construed as a limitation to the present invention.

[0042] In the description of the present invention, it should also be noted that unless otherwise clearly specified and limited, the terms "set", "installed", "connected", "connected" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and it can be the communication inside two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific situations.

[0043] A plasma exosome filtration and extraction device can batch-process and automate the exosome samples, further improving the efficiency of exosome separation in the laboratory.

[0044] Please refer to Figure 1 , the present invention provides a technical solution: a plasma exosome filtration and extraction device, including:

[0045] A low-temperature centrifuge 1 is arranged at the front end of the filtration and extraction device and is used for low-temperature centrifugation of the pre-treated samples after thawing and balancing. A dilution dispenser 2 is arranged on one side of the low-temperature centrifuge 1;

[0046] The dilution dispenser 2 can collect the supernatant of the sample tube after low-temperature centrifugation and can add PBS diluent for dilution;

[0047] A shaker 3 is used for shaking and mixing the diluted samples;

[0048] A filter 4 is used for filtering the diluted, shaken and homogenized samples;

[0049] An exosome collection box 5 can be connected to the output end of the filter 4 through multiple connectors.

[0050] Please refer to Figure 2 Figure 2 , a dilution dispenser 2 includes a dispensing bin 201, a shaker (not shown in the figure) disposed at the bottom side of the dispensing bin 201, a dilution tube 202 disposed in the dispensing bin 201, an input pump head, an output pump head (not shown in the figure), and an input suction tube and an output suction tube 203 respectively connected to the input pump head and the output pump head; the end of the input suction tube is branched into a plurality of input heads, and the plurality of input heads can extend into the upper layer of the sample tube after the sample is centrifuged by the low-temperature centrifuge 1 to collect the supernatant into the dispensing bin 201; the dilution tube 202 can be connected to an external PBS diluent, and the output side of the output suction tube 203 is connected to the shaker 3.

[0051] In this embodiment, the output suction tube 203 is connected to a dispensing rack 204 through a hose end, and the bottom side of the dispensing rack 204 is branched into a plurality of dispensing output heads 205. The number of the dispensing output heads 205 matches the number of the oscillation tube modules 311 on the oscillation tray 310, and the dispensing output heads 205 can extend into the oscillation tube modules 311.

[0052] In this embodiment, the above-mentioned dilution dispenser 2 can add PBS diluent for dilution, then use the shaker for pre-shaking, and then use the output suction tube 203 to divert the diluted sample into a plurality of oscillation tube modules 311 of the shaker 3, and the shaker 3 performs an oscillation operation on the oscillation tube modules 311. The advantage of the dilution dispenser 2 is that it can quickly transfer a large number of samples in the low-temperature centrifuge 1 and realize sample dilution through pre-shaking and introducing PBS.

[0053] Please refer to Figure 3 Figure 3 , in this embodiment, the shaker 3 includes a frame 301, an upper frame 302, a swing-type oscillation assembly, an oscillation tray 310 and an oscillation tube module 311. Among them, the upper frame 302 is arranged along the end face of the frame 301, and one-way sliding frames 303 are arranged on both sides of the upper frame 302. The oscillation tray 310 can be connected in a guiding manner inside the upper frame 302; the swing-type oscillation assembly includes an oscillation arm 304, a swing plate 305, a disc 306, a motor A, a large pulley 307 and a small pulley 308. Among them, two groups of discs 306 are provided and are coaxially rotatably connected to both sides of the upper frame 302. A large pulley 307 is arranged at the rear end of one of the discs 306. The large pulley 307 is linked with the small pulley 308 through a belt, and the small pulley 308 is connected to the shaft end of the motor A; a slide plate 309 is arranged on the end face of the large pulley 307, a swing plate 305 is arranged in front of the large pulley 307, and a one-way groove slidably connected to the slide plate 309 is arranged on the swing plate 305. The lower end of the swing plate 305 is hinged to the frame 301, and the upper end of the swing plate 305 is hinged to the oscillation arm 304.

[0054] In this embodiment, a sliding shaft is arranged inside the one-word carriage 303. The end of the sliding shaft can be connected to the oscillating tray 310, and the end of the oscillating arm 304 can be hinged to the sliding shaft. The oscillating arm 304 can drive the oscillating tray 310 to move back and forth when the turntable rotates.

[0055] Please refer to Figure 3 - Figure 4 , in this embodiment, a plurality of slots are arranged in an array on the oscillating tray 310, and an oscillating tube module 311 is placed in each slot. The oscillating tube module 311 includes a tube body 311a, an upper cover plate 311b and a confluence tube 311c. An upper cover plate 311b is arranged at the upper end of the tube body 311a, and an outlet tube is connected to the bottom side of the tube body 311a. An electromagnetic one-way valve 311d is arranged on the outlet tube. The ends of the outlet tubes of the plurality of oscillating tube modules 311 are connected to the confluence tube 311c. The confluence tube 311c is a flexible tube, and the end of the confluence tube 311c is divided into three pipelines and connected to the filter 4, and a one-way valve is arranged on each pipeline.

[0056] Please refer to Figure 5 , in this embodiment, there are at least three syringe-type prefilters. The three syringe-type prefilters are located on one side of the pushing module and are fixed at the bottom through a bracket. The syringe-type prefilter includes a syringe body 401, a piston push plate 402 and a prefilter layer. The prefilter layer is arranged on the front side of the syringe body 401, and the piston push plate 402 is arranged inside the syringe body 401. The rear end of the piston push plate 402 extends out of the syringe body 401 and is close to the pushing module.

[0057] In this embodiment, the pushing module includes a T-shaped bracket 403, a track 404, a rack push plate 405, a gear set 406 and a driving unit. A track 404 is arranged at the bottom side of the T-shaped bracket 403, and a driving unit is installed on the T-shaped bracket 403. The rack push plate 405 can be slidably connected to the track 404, and the front end of the rack push plate 405 is close to the piston push plate 402; the driving unit includes a motor B installed on the upper side of the T-shaped bracket 403 and a gear set 406 linked by a belt and a pulley. The gear set 406 includes three gears connected coaxially. The three gears are respectively engaged with the rack push plate 405 and push the rack push plate 405 to slide along the track 404 to push the piston push plate 402; the filter tube 407 is connected to the front end of the syringe body 401 through a joint tube, and multiple filter layers for filtering exosome samples are arranged in the filter tube 407.

[0058] In this embodiment, the above-mentioned filter 4 is based on the combination of the syringe-type prefilter and the filter tube 407. The syringe-type prefilter can be automatically filtered by filling a filter layer in a disposable syringe and automatically pushing it through the pushing module. The sample enters the filter tube 407 for deep filtration after passing through the syringe-type prefilter. Since the pushing module can pressurize the disposable syringe, the sample after entering the filter tube 407 has pressure and can achieve power-free filtration, and the impurity removal speed is fast.

[0059] An embodiment of the present invention also provides a method for using the above-mentioned plasma exosome filtration and extraction device, and the specific steps include:

[0060] S1: Take out the frozen sample, place it on ice for thawing. After the sample is completely thawed, balance the EP tube, and then place the sample in a low-temperature centrifuge 1 for low-temperature centrifugation to remove large vesicles;

[0061] S2: Use the input pump head and input suction tube of the dilution dispenser 2 to suck and collect the sample supernatant into the distribution chamber 201. The distribution chamber 201 adds PBS diluent through the dilution tube 202 for dilution, then use the shaker for pre-shaking, and then use the output suction tube 203 to divert the diluted sample into multiple oscillation tube modules 311 of the oscillator 3;

[0062] S3: Multiple oscillation tube modules 311 can achieve the oscillation effect by moving the oscillation tray 310 back and forth. After multiple reciprocating oscillations, the electromagnetic one-way valve 311d at the bottom of the tube body 311a is opened, and the samples in multiple tube bodies 311a converge through the confluence tube 311c into the syringe-type pre-filter;

[0063] S4: The syringe-type pre-filter can use the gear to engage with the rack push plate 405 and the piston push plate 402. The piston push plate 402 can filter and push out the sample entering the syringe body 401, so that the sample enters the filter tube 407 at high speed for filtration to remove impurities. The filtered sample is collected by the exosome collection box 5 and completed the exosome filtration and extraction after ultracentrifugation.

[0064] It should be noted that: the entire device is controlled by the total control system. Since the devices matched by the control system are common devices and belong to the existing mature technologies, the electrical connection relationship and the specific circuit structure are not described in detail here.

[0065] Although the embodiments of the present invention have been shown and described, for those of ordinary skill in the art, it can be understood that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principle and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A plasma exosome filtration and extraction device, characterized in that, Including: A low-temperature centrifuge (1), arranged at the front end of the filtration and extraction device and used for low-temperature centrifugation of the pre-treated samples after thawing and balancing. A dilution dispenser (2) is arranged on one side of the low-temperature centrifuge (1); The dilution dispenser (2) includes a distribution bin (201), a shaker arranged at the bottom side of the distribution bin (201), a dilution tube (202) arranged in the distribution bin (201), an input pump head, an output pump head, and an input suction tube and an output suction tube (203) respectively connected to the input pump head and the output pump head; A shaker (3), used for shaking and mixing the diluted samples. The shaker (3) includes a frame (301), an upper frame (302), a swing-type shaking assembly, a shaking tray (310) and a shaking tube module (311). Among them, the upper frame (302) is arranged along the end face of the frame (301), linear slides (303) are arranged on both sides of the upper frame (302), and the shaking tray (310) is connected to the inside of the upper frame (302) in a guiding manner; A filter (4), used for filtering the diluted, shaken and homogenized samples. The filter (4) includes a syringe-type pre-filter, a filter tube (407) and a pushing module. Among them, there are at least three groups of syringe-type pre-filters. The three groups of syringe-type pre-filters are located on one side of the pushing module and are fixed at the bottom side through a bracket. The syringe-type pre-filter includes a syringe body (401), a piston push plate (402) and a pre-filter layer; An exosome collection box (5), connected to the output end of the filter tube (407) through a plurality of connectors.

2. The plasma exosome filtration and extraction device according to claim 1, characterized in that: The end of the input suction tube is branched into a plurality of input heads, and the plurality of input heads extend into the upper layer of the sample tube after the low-temperature centrifuge (1) completes sample centrifugation to collect the supernatant into the distribution bin (201); the dilution tube (202) is connected to the external PBS diluent, and the output side of the output suction tube (203) is connected to the shaker (3).

3. The plasma exosome filtration and extraction device according to claim 1, characterized in that: The swing-type shaking assembly includes a shaking arm (304), a swing plate (305), a disc (306), a motor A, a large pulley (307) and a small pulley (308). Among them, there are two groups of discs (306) which are coaxially rotatably connected to both sides of the upper frame (302). A large pulley (307) is arranged at the rear end of one of the discs (306), and the large pulley (307) is linked with the small pulley (308) through a belt. The small pulley (308) is connected to the shaft end of the motor A; a slide plate (309) is arranged on the end face of the large pulley (307), a swing plate (305) is arranged in front of the large pulley (307), a linear slot for sliding connection with the slide plate (309) is arranged on the swing plate (305), the lower end of the swing plate (305) is hinged to the frame (301), and the upper end of the swing plate (305) is hinged to the shaking arm (304).

4. The plasma exosome filtration and extraction device according to claim 1, wherein: A slide shaft is arranged in the linear slide (303), the end of the slide shaft is connected to the shaking tray (310), the end of the shaking arm (304) is hinged to the slide shaft, and the shaking arm (304) drives the shaking tray (310) to move back and forth when the turntable rotates.

5. The plasma exosome filtration and extraction device according to claim 4, characterized in that: A plurality of slot holes are arranged in an array on the oscillating tray (310), and an oscillating tube module (311) is placed in each slot hole. The oscillating tube module (311) includes a tube body (311a), an upper cover plate (311b) and a confluence tube (311c). An upper cover plate (311b) is arranged at the upper end of the tube body (311a), an outlet tube is connected to the bottom side of the tube body (311a), and an electromagnetic one-way valve (311d) is arranged on the outlet tube. The ends of the outlet tubes of a plurality of oscillating tube modules (311) are connected to the confluence tube (311c). The confluence tube (311c) is a flexible tube, and the end of the confluence tube (311c) is connected to the filter (4) through three pipelines, and a one-way valve is arranged on each pipeline.

6. The plasma exosome filtration and extraction device according to claim 1, wherein: A pre-filter layer is arranged on the front side of the syringe body (401), and a piston push plate (402) is arranged inside the syringe body (401). The rear end of the piston push plate (402) extends out of the syringe body (401) and is close to the pushing module.

7. A plasma exosome filtration and extraction device according to claim 6, characterized in that: The pushing module includes a T-shaped bracket (403), a track (404), a rack push plate (405), a gear set (406) and a driving unit. A track (404) is arranged on the bottom side of the T-shaped bracket (403), a driving unit is installed on the T-shaped bracket (403), the rack push plate (405) is slidably connected to the track (404), and the front end of the rack push plate (405) is close to the piston push plate (402).

8. A plasma exosome filtration and extraction device according to claim 7, characterized in that: The driving unit includes a motor B installed on the upper side of the T-shaped bracket (403) and a gear set (406) linked by a belt and a pulley. The gear set (406) includes three gears connected coaxially. The three gears are respectively engaged with the rack push plate (405) and push the rack push plate (405) to slide along the track (404) to push the piston push plate (402); the filter tube (407) is connected to the front end of the syringe body (401) through a connecting tube, and a plurality of filter layers for filtering exosome samples are arranged in the filter tube (407).

9. The plasma exosome filtration and extraction device according to claim 1, characterized in that: The output suction tube (203) is connected to the distribution rack (204) through the end of a flexible tube. The bottom side of the distribution rack (204) is branched into a plurality of distribution output heads (205). The number of the distribution output heads (205) matches the number of the oscillating tube modules (311) on the oscillating tray (310), and the distribution output heads (205) extend into the oscillating tube modules (311).

10. The method of using a plasma exosome filtration and extraction device according to any one of claims 1-9, characterized in that: Including steps: S1: Take out the frozen sample, thaw it on ice. After the sample is completely thawed, balance the EP tube, and then place the sample in a low-temperature centrifuge (1) for low-temperature centrifugation to remove large vesicles; S2: Use the input pump head and input suction tube of the dilution dispenser (2) to suck and collect the sample supernatant into the distribution bin (201). The distribution bin (201) is diluted by adding PBS diluent through the dilution tube (202), then pre-shake it with a shaker, and then use the output suction tube (203) to divert the diluted sample into a plurality of oscillating tube modules (311) of the oscillator (3); S3: Multiple oscillation tube modules (311) can achieve an oscillation effect by moving back and forth using an oscillation tray (310). After multiple reciprocating oscillations, the electromagnetic one-way valve (311d) at the bottom of the tube body (311a) opens, and the samples in multiple tube bodies (311a) converge through a confluence tube (311c) and flow into a syringe-type pre-filter; S4: The syringe-type pre-filter can utilize the meshing of a gear and a rack push plate (405) and a piston push plate (402). The piston push plate (402) can filter and push out the samples entering the syringe body (401), enabling the samples to enter the filter tube (407) at high speed for filtration to remove impurities. The filtered samples are collected through an exosome collection box (5) and completed exosome filtration and extraction after ultracentrifugation.

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