Cell exosome extraction device and extraction method

Abstract

The invention is suitable for the technical field of cell exosome extraction, and provides a cell exosome extraction device and method, the cell exosome extraction device comprises a base box, the top of the base box is provided with a centrifugal box, the centrifugal box is internally provided with a rotatable centrifugal seat, and the centrifugal seat is used for placing a centrifugal test tube containing a cell sample solution; the filter shell is arranged above the base box and located on the side edge of the centrifugal box, the filter shell can transversely move, a valve type filter disc capable of being rotationally adjusted is arranged in the filter shell, and the valve type filter disc is used for filtering and treating a centrifuged cell sample solution. According to the cell exosome extraction device and the extraction method provided by the scheme, the problem that the extraction efficiency is affected due to the fact that the steps of cleaning, dismounting and mounting a filter inner core of an existing cell exosome extraction device are tedious and time and labor are consumed is solved.

Description

Technical Field

[0001] The present invention belongs to the technical field of extracellular vesicle extraction, and in particular relates to an extracellular vesicle extraction device and an extraction method. Background Art

[0002] Exosomes are small vesicles with a diameter of 30-150nm that are released by multivesicular bodies through exocytosis. They contain components such as proteins, nucleic acids, and lipids. They play a key role in the occurrence and development of diseases such as tumors, and have great potential in disease diagnosis and treatment.

[0003] At present, in biological research, the centrifugation and filtration method is commonly used for extracellular vesicle extraction, and laboratory centrifuges play an important role. However, existing extracellular vesicle filtration devices also have defects. The filter core is fixed and difficult to clean, and there are many residues and impurities that are easily introduced, which reduces the filtration effect. Taking the device with patent number CN15197827B as an example, after the extraction is completed, the disassembly, cleaning and installation steps of the filtration tube, filtration core and other components are cumbersome, which is time-consuming and labor-intensive, and the extraction quality and efficiency are easily affected by improper operation. Summary of the invention

[0004] The present invention provides an extracellular vesicle extraction device and an extraction method, aiming to solve the problems in the above-mentioned background technology that the cleaning, disassembly and installation steps of the filter inner core of the existing extracellular vesicle extraction device are cumbersome, time-consuming and labor-intensive, affecting the extraction efficiency.

[0005] To solve the above problems, the present invention is implemented as follows: an extracellular vesicle extraction device comprises: a base box, a centrifugal box is arranged on the top of the base box, a rotatable centrifugal seat is arranged in the centrifugal box, and the centrifugal seat is used to place a centrifugal test tube containing a cell sample solution; a filter shell arranged above the base box and located on the side of the centrifugal box, the filter shell can move laterally, and a rotatable and adjustable valve-type filter disc is arranged in the filter shell, and the valve-type filter disc is used to filter and process the cell sample solution after centrifugation; a cleaning box assembled on the top of the base box, the cleaning box is located on one side of the lateral movement track of the filter shell and has an inlet and outlet for the filter shell to enter and exit, a flushing pipe is fixedly installed in the cleaning box, which is used to spray cleaning liquid to flush the filter shell and the valve-type filter disc located below after the filter shell enters the cleaning box; a third air duct fixedly connected to the cleaning box is used to introduce hot air to dry the filter shell and the valve-type filter disc; an adjusting mechanism arranged on the cleaning box for regulating the lateral movement of the filter shell; and a cooling mechanism arranged in the base box and the centrifugal box, which is used to supply air and cool the inside of the centrifugal box.

[0006] Preferably, the adjustment mechanism includes: a slide plate fixed on the top of the cleaning box, a baffle plate slidably sleeved on the slide plate to cover the inlet and outlet of the filter shell after the filter shell enters the cleaning box, a connecting frame fixedly mounted on the baffle plate, a placement frame for placing the filter shell fixedly mounted on the connecting frame, and the connecting frame and the placement frame with the filter shell placed thereon can both follow the movement of the baffle plate to enter and exit along the inlet and outlet of the cleaning box.

[0007] Preferably, the cooling mechanism includes: a plurality of Peltier cooling plates mounted on the inner wall of the centrifugal box for cooling; an air supply box fixed in the base box, a mounting rod provided in the air supply box, and fan blades mounted on the mounting rod; a transmission rod rotatably mounted on the air supply box, one end of the transmission rod being fixedly connected to the mounting rod via a coupling; and a first air duct fixedly connected to the air supply box, the air outlet end of the first air duct being connected to the centrifugal box.

[0008] Preferably, a placement seat located between the centrifuge box and the cleaning box is provided on the top of the base box, and at least two beakers are provided on the placement seat. The beakers are placed on the movement trajectory of the filter shell so that the beakers can receive the solution filtered by the filter shell. The filter shell is fixedly connected with a liquid outlet pipe, and the liquid outlet pipe is used to guide the supernatant filtered in the filter shell to the corresponding beaker.

[0009] Preferably, the bottom of the cleaning box is fixedly connected to a drain pipe for discharging dirty liquid, the liquid outlet end of the drain pipe extends into the base box, and a collecting box is provided in the base box, and the collecting box is used to receive and collect the dirty liquid discharged from the drain pipe.

[0010] Preferably, one side of the centrifugal box is fixedly connected with an exhaust pipe and a drain pipe, the exhaust pipe is used for exhausting heat, and the drain pipe is used for discharging condensed water.

[0011] Preferably, a detachable cover is provided on the top of the centrifuge box, a handle for grasping is fixedly mounted on the cover, and a liquid supply pipe is fixedly mounted on the liquid inlet end of the flushing pipe, and the liquid supply pipe is used to connect to a cleaning liquid supply device.

[0012] Preferably, a placement groove and a plurality of air holes are provided on the centrifugal seat, a second spring is symmetrically fixedly mounted on the inner wall of the placement groove, and a clamping block for clamping the centrifugal test tube is fixedly mounted on the second spring.

[0013] Preferably, a limiting ring is fixedly mounted on the filter shell, a plug-in block is fixedly mounted on the bottom of the limiting ring, a groove matching the plug-in block is provided on the placement rack, a second rotating shaft is rotatably mounted on the filter shell through a bearing, and the second rotating shaft is fixedly connected to the valve-type filter disc.

[0014] The present invention also provides an extraction method of an extracellular vesicle extraction device, comprising the above-mentioned extracellular vesicle filtration extraction device, and the extraction method comprises the following steps: S1: Sample preparation: Select the liquid of the cell solution for biological culture to obtain the sample solution. Take a certain amount of sample solution, add it into the centrifuge test tube, place the centrifuge test tube in the placement slot of the centrifuge seat, and clamp the centrifuge test tube with the clamp under the action of the second spring; S2: First centrifugation: Close the cover on the top of the centrifuge box, set the speed and centrifugation time through the control module, start the rotation of the centrifuge seat, and perform the first centrifugation operation on the sample solution. During this process, the Peltier refrigeration sheet on the inner wall of the centrifuge box works to cool the high-speed rotating centrifuge seat and centrifuge test tube; at the same time, the fan blades of the air supply box in the base box rotate, and the air is supplied to the centrifuge box through the first air duct for cooling, the exhaust pipe exhausts the heat, and the drain pipe discharges the condensed water; S3: First filtration: After the first centrifugation is completed, pour the liquid after centrifugation in the centrifuge test tube into the filter shell, and the liquid is filtered through the valve filter. The supernatant after filtration enters the beaker through the outlet pipe to remove the sediment, collect the supernatant, and remove living cells and larger impurities. ; S4: Cleaning operation: Use the adjustment mechanism to move the filter shell into the cleaning box, and use the flushing pipe to spray the cleaning liquid to rinse the filter shell and the valve filter disc. The cleaned dirty liquid is discharged into the collection box through the drain pipe; S5: Second centrifugation: Add the supernatant in the beaker to the centrifuge tube again, set the centrifugation time through the control module again, and perform the second centrifugation operation. During the centrifugation process, the cooling components of the Peltier cooling plate and the air supply box work synchronously; S6: Second filtration: After the second centrifugation is completed, repeat the first filtration operation to obtain the supernatant again, and this time the dead cells and cell debris are removed; S7: Third centrifugation and exosome preparation: Add the supernatant obtained from the second filtration to the centrifuge tube again, set the centrifugation time through the control module again, perform strong centrifugation, and then pour the liquid in the centrifuge tube into the filter shell for filtration. This time, remove the supernatant to obtain a precipitate. The precipitate is resuspended in μL PBS for purification and extraction to obtain cell exosomes.

[0015] Compared with the related art, the cell extracellular vesicle extraction device and extraction method provided by the present invention have the following beneficial effects: Compared with the prior art, the cell exosome extraction device provided in this solution; Through standardized sample preparation, centrifugation, filtration, washing and re-centrifugation steps, the cell exosomes are gradually purified to obtain high-quality products. The laterally adjustable filter shell cooperates with the washing box and the adjustment mechanism to facilitate cleaning, simplify the process, reduce operating losses, and improve extraction efficiency. The cooling mechanism in the base box and the centrifuge box solves the problem of heat generation during high-speed rotation, ensures the extraction environment of cell exosomes, improves quality and experimental reliability, and extends the life of the equipment. The placement seat, beaker and liquid outlet pipe of the filter shell on the top of the base box are precisely connected to achieve accurate collection of filtrate, improve efficiency and basic quality of subsequent experiments. The drain pipe of the washing box and the collection box in the base box effectively collect dirty liquid, keep the device clean, and ensure the cleaning effect and extraction quality. The exhaust pipe and drain pipe of the centrifuge box exhaust heat and drain water to avoid high-temperature water accumulation in the box, ensuring stable operation of the equipment and extraction quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is a schematic diagram of the main structure of an exosome extraction device and an extraction method provided by the present invention; Figure 2 It is a rear structural schematic diagram of an exosome extraction device and an extraction method provided by the present invention; Figure 3 It is a schematic diagram of the main cross-sectional structure of an exosome extraction device and an extraction method provided by the present invention; Figure 4 for Figure 3 An enlarged structural diagram of part A shown in FIG. Figure 5 for Figure 3 An enlarged schematic diagram of the structure of part B shown in FIG. Figure 6 for Figure 3 An enlarged schematic diagram of the structure of part C shown in FIG. Figure 7 for Figure 3 An enlarged structural diagram of part D shown in FIG. Figure 8 It is a structural schematic diagram of the centrifugal seat in the present invention; Fig. 9 It is a structural schematic diagram of the mesh plate in the present invention; Fig.10 It is a structural schematic diagram of the baffle and the connecting frame in the present invention; Fig.11 It is a schematic diagram of the structure of the placement rack in the present invention; Fig.12 It is a schematic diagram of the top view of the filter shell in the present invention.

[0017] 1. Base box; 2. Centrifugal box; 3. Centrifugal seat; 4. Peltier refrigeration plate; 5. Cleaning box; 6. Slide plate; 7. Baffle; 8. Connecting frame; 9. Placement frame; 10. Filter shell; 11. Valve filter; 12. Placement seat; 13. Beaker; 14. Flushing pipe; 15. Liquid supply pipe; 16. Air supply box; 17. Motor; 18. Transmission rod; 19. Rotating rod; 20. Gear sleeve; 21. Electric telescopic rod; 22. Assembly plate; 23. Gear cylinder; 24. Tooth groove block; 25. Tooth groove rod; 26. First bevel gear; 27. Second bevel gear; 28. First pulley; 29. ​​Second pulley; 30. First timing belt; 31. First air duct; 32. Mounting rod; 33. Fan blade; 34. First The second air duct; 35. the heating box; 36. the third air duct; 37. the electric heating tube; 38. the mesh plate; 39. the first rotating shaft; 40. the cleaning brush; 41. the activated carbon plate; 42. the solenoid valve; 43. the diverter plate; 44. the second rotating shaft; 45. the third bevel gear; 46. the first shaft rod; 47. the fourth bevel gear; 48. the second shaft rod; 49. the third pulley; 50. the second timing belt; 51. the fifth bevel gear; 52. the drain pipe; 53. the collecting box; 54. the cover plate; 55. the exhaust pipe; 56. the drain pipe; 57. the pin rod; 58. the first spring; 59. the positioning block; 60. the limit block; 61. the test tube; 62. the second spring; 63. the clamping block; 64. the connecting rod; 65. the liquid outlet pipe; 66. the partition. DETAILED DESCRIPTION

[0018] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by technicians in the technical field of the present application; the terms used in the specification of the application herein are only for the purpose of describing specific embodiments and are not intended to limit the present application; the terms "including" and "having" in the specification and claims of the present application and the above-mentioned figure descriptions and any variations thereof are intended to cover non-exclusive inclusions. The terms "first", "second", etc. in the specification and claims of the present application or the above-mentioned figures are used to distinguish different objects, rather than to describe a specific order; the terms "inside", "outside", "left", and "right" indicate directions or positional relationships based on the directions or positional relationships shown in the drawings, which are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction, and therefore cannot be understood as a limitation on the present invention.

[0019] Reference to "embodiments" herein means that a particular feature, structure, or characteristic described in conjunction with the embodiments may be included in at least one embodiment of the present application. The appearance of the phrase in various locations in the specification does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment that is mutually exclusive with other embodiments. It is explicitly and implicitly understood by those skilled in the art that the embodiments described herein may be combined with other embodiments.

[0020] The embodiment of the present invention provides a device and method for extracting extracellular vesicles, such as Figure 1-12 As shown, the cell extracellular vesicle extraction device and extraction method include: a base box 1, a centrifugal box 2 is arranged on the top of the base box 1, a rotatable centrifugal seat 3 is arranged in the centrifugal box 2, and the centrifugal seat 3 is used to place a centrifugal test tube 61 containing a cell sample solution; a filter shell 10 is arranged above the base box 1 and located on the side of the centrifugal box 2, the filter shell 10 can move laterally, and a rotatable and adjustable valve-type filter disc 11 is arranged in the filter shell 10, and the valve-type filter disc 11 is used to filter and process the cell sample solution after centrifugation; a cleaning box 5 assembled on the top of the base box 1, the cleaning box 5 An inlet and outlet for the filter shell 10 to enter and exit is provided on one side of the lateral movement trajectory of the filter shell 10. A flushing pipe 14 is fixedly installed in the cleaning box 5, which is used to spray cleaning liquid to flush the filter shell 10 and the valve-type filter disc 11 located below after the filter shell 10 enters the cleaning box 5; a third air duct 36 fixedly connected to the cleaning box 5 is used to introduce hot air to dry the filter shell 10 and the valve-type filter disc 11; an adjustment mechanism is arranged on the cleaning box 5 for regulating the lateral movement of the filter shell 10; and a cooling mechanism is arranged in the base box 1 and the centrifugal box 2, which is used to supply air to cool the inside of the centrifugal box 2.

[0021] In this embodiment, when preparing the sample, the cell sample solution is loaded into the centrifuge test tube 61 and placed in the placement groove of the centrifuge seat 3 for clamping. Close the cover plate 54 of the centrifuge box 2, set the parameters and start the centrifuge seat 3 to rotate and centrifuge, and the Peltier cooling plate 4 in the centrifuge box 2 and the cooling component of the air supply box 16 in the base box 1 work synchronously. After the centrifugation is completed, the test tube liquid is poured into the filter shell 10, filtered through the valve filter 11, and the supernatant flows into the beaker 13. After filtration, the filter shell 10 is moved to the cleaning box 5 for cleaning by the adjustment mechanism, and the dirty liquid is discharged into the collection box 53. If purification is required, the supernatant can repeat the steps of centrifugation, filtration, and cleaning, and finally the precipitate is obtained by strong centrifugation and filtration, and the cell exosomes are extracted by resuspending with 100μL PBS.

[0022] The filter housing 10 of the present invention can be adjusted laterally, and is matched with a cleaning box 5 and an adjustment mechanism. Compared with the traditional device, it can be easily moved to the cleaning box 5 for cleaning, which simplifies the process, avoids tedious disassembly, reduces operating losses, and improves extraction efficiency.

[0023] A cooling mechanism is provided on the base box 1 and the centrifugal box 2, including a Peltier cooling sheet 4, an air supply box 16, etc., as well as an exhaust pipe 55 and a drain pipe 56. This mechanism effectively solves the problem of heat generation during high-speed rotation of the centrifuge, ensures that the cell exosomes are extracted at a suitable temperature, and protects their structure and components from being damaged by high temperature.

[0024] In a further preferred embodiment of the present invention, the adjusting mechanism includes: a slide plate 6 fixed on the top of the cleaning box 5, a baffle 7 is slidably sleeved on the slide plate 6 to cover the inlet and outlet of the filter shell 10 after entering the cleaning box 5, a connecting frame 8 is fixedly mounted on the baffle 7, and a placement frame 9 for placing the filter shell 10 is fixedly mounted on the connecting frame 8. The connecting frame 8 and the placement frame 9 with the filter shell 10 can both follow the movement of the baffle 7 and enter and exit the inlet and outlet of the cleaning box 5.

[0025] In this embodiment, when the filter shell 10 needs to be cleaned, first push the baffle 7 to slide on the slide plate 6. Since the baffle 7 is fixedly mounted on the connecting frame 8, and the placement frame 9 is assembled on the connecting frame 8, as the baffle 7 slides, the connecting frame 8 and the placement frame 9 will move synchronously. At this time, the filter shell 10 placed on the placement frame 9 will also move laterally until the filter shell 10 moves into the cleaning box 5. When the cleaning is completed, push the baffle 7 in the reverse direction to remove the filter shell 10 from the cleaning box 5 and restore it to the filtering working position. During cleaning, the baffle 7 can block the inlet and outlet of the cleaning box 5 to play a protective role; In traditional extracellular vesicle extraction devices, the filter core is fixed and difficult to clean, but the adjustment mechanism of the present invention has completely changed this situation. Through the simple operation of the sliding baffle 7, the filter shell 10 can be easily moved into the cleaning box 5, and the filter shell 10 and the valve-type filter disc 11 can be thoroughly cleaned by the flushing pipe 14. This design avoids complicated disassembly steps and greatly saves cleaning time and energy. For example, compared with the device of patent number CN15197827B, which requires multiple cumbersome steps to clean the filter component, the adjustment mechanism of the present invention makes the cleaning process simple and efficient, significantly improving the convenience of cleaning; The setting of the regulating mechanism optimizes the entire extracellular vesicle extraction process. It not only simplifies the cleaning process, but also reduces the risk of component damage caused by frequent disassembly and installation, thereby increasing the service life of the device. At the same time, efficient cleaning ensures the cleanliness of the filter housing 10 and the valve-type filter disc 11, so that each filtering operation can maintain a good effect, thereby improving the extraction quality of extracellular vesicles. In addition, due to the simplification of the cleaning process, the efficiency of the entire extraction process has been greatly improved, saving valuable time for scientific researchers and helping to promote the rapid development of extracellular vesicle-related research.

[0026] In a further preferred embodiment of the present invention, the cooling mechanism includes: a plurality of Peltier refrigeration plates 4 mounted on the inner wall of the centrifugal box 2 for cooling; an air supply box 16 fixed in the base box 1, a mounting rod 32 is provided in the air supply box 16, and a fan blade 33 is mounted on the mounting rod 32; a transmission rod 18 rotatably mounted on the air supply box 16, one end of the transmission rod 18 is fixedly connected to the mounting rod 32 through a coupling; a first air duct 31 fixedly connected to the air supply box 16, and the air outlet end of the first air duct 31 is connected to the centrifugal box 2.

[0027] In this embodiment, when the centrifugal seat 3 in the centrifugal box 2 drives the centrifugal test tube 61 to rotate at high speed, the cooling mechanism is activated. The Peltier cooling sheet 4 on the inner wall of the centrifugal box 2 works, and its cooling end cools the centrifugal seat 3 and the centrifugal test tube 61. At the same time, the air supply box 16 in the base box 1 operates, and the transmission rod 18 rotates to drive the installation rod 32 and the fan blade 33 to rotate, and the wind is sent into the centrifugal box 2 through the first air guide 31, taking away the heat, and achieving effective cooling.

[0028] In the traditional cell extracellular vesicle extraction device, the centrifuge rotates at high speed and generates heat that is difficult to dissipate, which affects the extraction quality. The cooling mechanism of the present invention has significant advantages. The Peltier cooling sheet 4 cools down from the heat source, and the air supply box 16 strengthens the air circulation and heat dissipation. The double cooling can accurately and efficiently control the temperature and avoid the temperature affecting the extraction quality.

[0029] The cooling mechanism optimizes the extracellular vesicle extraction process. The stable temperature environment ensures the extracellular vesicle structure and composition, improves the extraction quality, reduces experimental errors, and enhances experimental repeatability and reliability. It also extends the service life of the centrifuge box 2 and other components, reduces maintenance costs, provides strong support for scientific research, and promotes the development of extracellular vesicle research. In addition, the Peltier cooling sheet 4 is controlled by the controller, the end is cooled, and the heat conduction end dissipates heat outside the centrifuge box 2.

[0030] In a further preferred embodiment of the present invention, a placement seat 12 located between the centrifuge box 2 and the cleaning box 5 is provided on the top of the base box 1, and at least two beakers 13 are provided on the placement seat 12. The beakers 13 are placed on the movement trajectory of the filter shell 10 so that the beakers 13 can receive the solution filtered by the filter shell 10. The filter shell 10 is fixedly connected with a liquid outlet pipe 65, and the liquid outlet pipe 65 is used to guide the supernatant filtered in the filter shell 10 to the corresponding beaker 13.

[0031] In this embodiment, during the extraction of extracellular vesicles, when the centrifuged cell sample solution enters the filter housing 10 and is filtered by the valve filter 11, the supernatant will flow out through the liquid outlet pipe 65 fixedly connected to the filter housing 10. The position of the liquid outlet pipe 65 is carefully designed, and its outlet is just aligned with the beaker 13 placed on the placement seat 12. In this way, the filtered supernatant can flow smoothly into the beaker 13, realizing the accurate collection of the filtered solution, and the whole process is coherent and smooth.

[0032] Compared with the traditional extracellular vesicle extraction device, the solution collection design in the present invention has significant advantages. The precise docking of the liquid outlet tube 65 and the beaker 13 avoids the spillage and loss of the supernatant during the collection process, greatly improving the accuracy of the solution collection after filtration. At the same time, the fixed placement seat 12 and the matching liquid outlet tube 65 design eliminate the need for operators to perform tedious positioning and placement operations after each filtration, saving time and improving filtration efficiency. For example, in some traditional devices, the supernatant may be partially lost due to unstable placement or position deviation of the collection container, affecting subsequent experimental results, and the present invention solves these problems well; The coordinated design of the placement seat 12, the beaker 13 and the liquid outlet tube 65 has played a positive role in promoting the entire cell exosome extraction work. Accurate solution collection ensures that a sufficient amount of pure supernatant can be obtained in each experiment, which provides a good foundation for subsequent centrifugation and purification operations and helps to improve the extraction quality of cell exosomes. The efficient collection process also speeds up the progress of the entire extraction work, allowing researchers to complete multiple experiments in a shorter time and improve experimental efficiency. In addition, this stable and reliable collection method reduces the uncertainty in the experimental process and improves the repeatability and reliability of the experimental results.

[0033] In a further preferred embodiment of the present invention, the bottom of the cleaning box 5 is fixedly connected to a drain pipe 52 for discharging dirty liquid, and the liquid outlet end of the drain pipe 52 extends into the base box 1. A collecting box 53 is provided in the base box 1, and the collecting box 53 is used to receive and collect the dirty liquid discharged from the drain pipe 52.

[0034] In this embodiment, when the filter housing 10 and the valve-type filter disc 11 are cleaned, the flushing pipe 14 in the cleaning box 5 sprays the cleaning liquid to flush them. The dirty liquid generated by the flushing will flow downward under the action of gravity and be discharged through the drain pipe 52 fixedly connected to the bottom of the cleaning box 5. The liquid outlet end of the drain pipe 52 extends into the base box 1 and is aligned with the collection box 53 placed in the base box 1. In this way, the dirty liquid can flow smoothly into the collection box 53, realizing the effective collection of the dirty liquid generated during the cleaning process. The whole process is simple and efficient, and the drain pipe 52 can also be used for exhaust during the drying process.

[0035] Compared with the traditional extracellular vesicle extraction device, the dirty liquid discharge and collection design of the present invention has obvious advantages. The setting of the drain pipe 52 and the collection box 53 enables the dirty liquid after cleaning to be collected in a timely and orderly manner, avoiding the dirty liquid from remaining in the device or flowing around, maintaining the cleanliness of the inside of the device, and reducing the bacteria breeding and equipment corrosion problems that may be caused by the residual dirty liquid. At the same time, this integrated design eliminates the need for operators to manually clean the dirty liquid after each cleaning, greatly simplifies the operation process, and improves the convenience of operation. For example, in some traditional devices, the treatment of the dirty liquid after cleaning is relatively cumbersome and may require manual dumping, which is not only time-consuming and labor-intensive, but also prone to pollution. The present invention solves these problems well.

[0036] The coordinated design of the cleaning box 5, the drain pipe 52 and the collection box 53 plays an important supporting role in the entire extracellular vesicle extraction work. A good waste liquid collection system ensures the smooth progress of the cleaning work, thereby ensuring the cleanliness of the filter shell 10 and the valve filter 11, so that each filtering operation can maintain a good effect, which helps to improve the extraction quality of extracellular vesicles. The simplified waste liquid treatment process also improves the efficiency of the entire extraction work, saves the time and energy of scientific researchers, and allows them to focus more on the extraction and research of extracellular vesicles.

[0037] In a further preferred embodiment of the present invention, an exhaust pipe 55 and a drain pipe 56 are fixedly connected to one side of the centrifugal box 2, wherein the exhaust pipe 55 is used for exhausting heat, and the drain pipe 56 is used for discharging condensed water.

[0038] In this embodiment, when the extracellular vesicle extraction device is running, the centrifugal seat 3 in the centrifugal box 2 drives the centrifugal test tube 61 to rotate at high speed, which will generate a lot of heat. At this time, the Peltier cooling sheet 4 on the inner wall of the centrifugal box 2 works to cool down, and the air supply box 16 supplies air to the box through the first air duct 31. The hot air and condensed water generated in the process need to be discharged in time, and the exhaust pipe 55 is fixed on one side of the centrifugal box 2 to discharge hot air and dissipate heat, and the drain pipe 56 also discharges condensed water on one side to prevent water accumulation in the box.

[0039] Compared with the conventional device, the exhaust pipe 55 and the drain pipe 56 of the present invention have obvious design advantages. Continuous exhaust and heat dissipation prevent the centrifuge box 2 from being overheated, preventing high temperature from damaging the cell exosome samples and equipment components, and ensuring the stable operation of the equipment. The drain pipe 56 drains water in time to avoid condensed water accumulation causing short circuits, corrosion and other problems, thereby extending the service life of the equipment. The conventional device lacks effective heat dissipation and drainage measures and is prone to failure. The present invention solves these problems.

[0040] The coordinated design of the exhaust pipe 55 and the drain pipe 56 plays a key supporting role in the cell exosome extraction experiment. The stable temperature environment and dry internal environment ensure that the structure and composition of the cell exosomes are not affected during the extraction process, thereby improving the extraction quality of the cell exosomes.

[0041] In a further preferred embodiment of the present invention, a removable cover plate 54 is provided on the top of the centrifuge box 2, a handle for grasping is fixedly mounted on the cover plate 54, and a liquid supply pipe 15 is fixedly mounted on the liquid inlet end of the flushing pipe 14, and the liquid supply pipe 15 is used to connect to a cleaning liquid supply device.

[0042] In this embodiment, when the cell exosomes are extracted, if the inside of the centrifuge box 2 needs to be operated, such as taking and placing the centrifuge tube 61, the researcher can easily remove the detachable cover plate 54 by grasping the handle on the cover plate 54, and then put it back after the operation to ensure that the centrifuge box 2 is closed during operation. When cleaning the filter housing 10 and the valve-type filter disc 11, the liquid supply pipe 15 is connected to the cleaning liquid supply device, and the cleaning liquid enters the flushing pipe 14 through the liquid supply pipe 15 for spray cleaning.

[0043] Compared with the traditional device, this part of the design of the present invention has significant advantages. The removable cover plate 54 with a handle on the top of the centrifugal box 2 makes the operation more convenient, and the centrifugal box 2 can be opened and closed without additional tools, which reduces the operation time. The flushing pipe 14 is connected to the cleaning liquid supply device through the liquid supply pipe 15 to ensure the stable and uniform supply of the cleaning liquid, improve the cleaning effect, ensure the cleanliness of the filter components, and provide good conditions for subsequent extraction.

[0044] The detachable cover 54 of the centrifuge box 2 and the coordinated design of the flushing tube 14 and the liquid supply tube 15 play an important role in promoting the extraction of extracellular vesicles. The convenient operation of the centrifuge box is conducive to the rapid replacement of samples and improves the experimental efficiency. The good cleaning effect ensures the performance of the filter component, which in turn helps to improve the extraction quality of extracellular vesicles.

[0045] In a further preferred embodiment of the present invention, a placement groove and a plurality of air holes are provided on the centrifugal seat 3, a second spring 62 is symmetrically fixedly mounted on the inner wall of the placement groove, and a clamping block 63 for clamping the centrifugal test tube 61 is fixedly mounted on the second spring 62.

[0046] In this embodiment, when performing an exosome extraction experiment, a centrifugal test tube 61 containing a cell sample solution is placed in the placement groove of the centrifugal seat 3. The second spring 62 on the inner wall of the placement groove drives the clamp 63 to clamp the centrifugal test tube 61 to ensure its stability when the centrifugal seat 3 rotates at high speed. During centrifugation, the air holes on the centrifugal seat 3 cooperate with the cooling mechanism to enhance air circulation and heat dissipation.

[0047] Compared with the traditional centrifugal device, the centrifugal seat 3 of the present invention has obvious design advantages. The second spring 62 and the clamp 63 can adapt to test tubes of different sizes, and can be fixed reliably to avoid shaking, displacement or falling off, thereby improving the safety of the experiment. The pores optimize air circulation, allowing the cold air of the cooling mechanism to circulate better, reduce the temperature of the test tube, ensure that the sample is centrifuged in a suitable environment, and improve the quality of cell exosome extraction.

[0048] The coordinated design of the placement groove, air hole, second spring 62 and clamp block 63 of the centrifugal seat 3 brings positive effects. Stable fixation saves experimental preparation time, good heat dissipation ensures the properties of the sample solution, and helps to obtain purer and more complete cell exosomes.

[0049] In a further preferred embodiment of the present invention, a limiting ring is fixedly provided on the filter shell 10, a plug-in block is fixedly installed on the bottom of the limiting ring, a groove adapted to the plug-in block is provided on the placement rack 9, and a second rotating shaft 44 is rotatably installed on the filter shell 10 through a bearing, and the second rotating shaft 44 is fixedly connected to the valve type filter 11.

[0050] In this embodiment, during the extracellular vesicle extraction process, when the filter shell 10 needs to be placed on the placement rack 9, the plug-in block at the bottom of the limiting ring fixed on the filter shell 10 is aligned with the corresponding groove on the placement rack 9 and then inserted into it, so that the filter shell 10 can be quickly and accurately positioned on the placement rack 9.

[0051] Compared with the traditional filtering device, the present invention has significant advantages in the design of the filter housing 10. The cooperation between the limit ring and the plug-in block and the groove of the placement frame 9 realizes the precise positioning of the filter housing 10, avoids the leakage of the solution caused by unstable placement or position deviation, and improves the stability and reliability of the filtering process. At the same time, the second rotating shaft 44 rotatably mounted by the bearing is fixedly connected to the valve-type filter disc 11, so that the valve-type filter disc 11 can be flipped for subsequent cleaning to make it clean more thoroughly.

[0052] In order to further improve the use effect of the device, in addition to the above scheme, this scheme also has the following embodiments: In another embodiment of the present invention, a driving mechanism for driving the centrifugal seat 3 and the transmission rod 18 to rotate is provided in the base box 1, and the driving mechanism includes: a motor 17 fixedly installed in the base box 1, and the output shaft of the motor 17 is fixedly connected to the transmission rod 18 through a coupling; a rotating rod 19 rotatably installed in the base box 1, and a gear sleeve 20 is fixedly installed on one end of the rotating rod 19, and a detachable tooth groove block 24 is meshed in the gear sleeve 20; an electric telescopic rod 21 fixedly installed in the base box 1, and an assembly plate 22 is fixedly installed on the output rod of the electric telescopic rod 21; a gear cylinder 23 rotatably installed on the assembly plate 22 through a bearing, and the gear cylinder 23 One end is fixedly connected to the tooth groove block 24; the tooth groove rod 25 meshing in the gear cylinder 23; the connecting rod 64 rotatably mounted on the base box 1 through a bearing, the top end of the connecting rod 64 extends into the centrifugal box 2 and is fixedly connected to the centrifugal seat 3 through a cross seat; the first bevel teeth 26 and the second bevel teeth 27 for transmission are respectively fixedly mounted on the tooth groove rod 25 and the bottom end of the connecting rod 64, the first bevel teeth 26 meshing with the second bevel teeth 27; the first pulley 28 and the second pulley 29 are respectively fixedly mounted on the transmission rod 18 and the rotating rod 19, and the first pulley 28 and the second pulley 29 are provided with a first timing belt 30 for transmission.

[0053] In this embodiment, when the extracellular vesicle extraction device is started, the motor 17 in the base box 1 works, and its output shaft drives the transmission rod 18 to rotate through the coupling, and the first pulley 28 on the transmission rod 18 rotates accordingly, and the second pulley 29 and the rotating rod 19 are driven to rotate through the first timing belt 30, and the gear sleeve 20 at one end of the rotating rod 19 also rotates accordingly. If the centrifugal seat 3 is to be driven, the electric telescopic rod 21 is started, which pushes the assembly plate 22 to make the tooth groove block 24 at one end of the gear cylinder 23 mesh with the gear sleeve 20, thereby driving the gear cylinder 23 to rotate, and the gear cylinder 23 drives the tooth groove rod 25 to move, and the first bevel teeth 26 on the tooth groove rod 25 mesh with the second bevel teeth 27 at the bottom end of the connecting rod 64, driving the connecting rod 64 to rotate, and finally realizing the rotation of the centrifugal seat 3.

[0054] Compared with the traditional driving method, the driving mechanism of the present invention has obvious advantages. Multiple components work together to achieve efficient transmission and flexible distribution of the power of the motor 17, which can stably transmit power to the centrifugal seat 3 and the transmission rod 18, ensuring stable and reliable operation of the equipment. In addition, the electric telescopic rod 21 cooperates with the gear sleeve 20 and the tooth groove block 24 to flexibly control the drive of the centrifugal seat 3. The tooth groove block 24 and the gear sleeve 20 can be separated when not needed to avoid energy consumption and component wear.

[0055] The driving mechanism is key to improving the performance of the extracellular vesicle extraction device. Stable power transmission ensures the stability of the centrifugal seat 3 during high-speed rotation, which helps to improve the centrifugal effect and extraction quality of extracellular vesicles. The flexible operation mode allows researchers to accurately control the operating state of the centrifugal seat 3 and improve the accuracy and repeatability of the experiment.

[0056] In another embodiment of the present invention, the air supply box 16 is provided with an air supply mechanism for supplying hot air into the third air duct 36, and the air supply mechanism includes: a second air duct 34 fixedly connected to the air supply box 16, and the air outlet end of the second air duct 34 is fixedly connected to the heating box 35; a plurality of partitions 66 fixedly installed in the heating box 35 for separating spaces, and ventilation holes are opened on adjacent partitions 66; and a plurality of electric heating tubes 37 arranged in the heating box 35 for heating.

[0057] In this embodiment, when hot air is to be supplied to the third air duct 36, the air in the air supply box 16 enters the heating box 35 through the second air duct 34. In the heating box 35, the air flows in a zigzag manner in the space separated by the multiple partitions 66, and the temperature controller controls the multiple electric heating tubes 37 to heat the air, and the heated hot air flows into the third air duct 36, thereby realizing the hot air supply to a specific area.

[0058] Compared with the traditional temperature control method, the air supply mechanism of the present invention has significant advantages. Multiple partitions 66 extend the air residence time, allowing it to fully contact the electric heating tube 37, and the heating is more uniform and effective. The multiple electric heating tubes 37 work together to accurately control the temperature, meet the requirements of hot air temperature for different experiments, avoid abnormal temperature affecting the extraction of extracellular vesicles, and flexibly adjust the local temperature of the device.

[0059] The air supply mechanism has a significant effect on the extraction of extracellular vesicles. Precise temperature control ensures that the structure and composition of extracellular vesicles are not damaged by temperature fluctuations, thereby improving the quality of extraction. Flexible hot air supply provides more possibilities for experiments, expands the scope of experimental application, enhances the functionality and practicality of the device, improves the overall performance of the equipment, provides strong technical support for extracellular vesicle research, and promotes the development of extraction technology.

[0060] In another embodiment of the present invention, a filtering mechanism for filtering impurities is provided on the air supply box 16, and the filtering mechanism includes: a mesh plate 38 fixedly installed on the air inlet of the air supply box 16 by bolts, and the mesh plate 38 is used to filter large impurities and flying insects; a first rotating shaft 39 rotatably installed on the mesh plate 38 through a bearing, and a cleaning brush 40 for cleaning the mesh plate 38 is fixedly installed on the first rotating shaft 39; and a detachable activated carbon plate 41 is provided in the air supply box 16, and the activated carbon plate 41 is used to filter air.

[0061] In this embodiment, when the air supply box 16 is working, outside air enters from the air inlet. The mesh plate 38 at the air inlet initially filters the air, blocks large impurities and flying insects, and ensures that the incoming air is free of large particle pollutants. When the air passes through the mesh plate 38, the cleaning brush 40 on the first rotating shaft 39 rotates to clean the mesh plate 38 to prevent impurities from accumulating and affecting the filtering effect. The air after the initial filtration enters the air supply box 16, and then passes through the detachable activated carbon plate 41. The activated carbon plate 41 uses adsorption to filter odors, tiny particles and harmful gases, and the purified air is sent to the subsequent process.

[0062] Compared with the traditional air supply filtering method, the filtering mechanism of the present invention has obvious advantages. The mesh plate 38 intercepts large impurities at the source, reducing the risk of equipment damage. The cleaning brush 40 ensures the filtering performance of the mesh plate 38 and saves maintenance time. The detachable activated carbon plate 41 deeply purifies the air and is easy to replace and maintain, ensuring a good filtering effect.

[0063] The filtration mechanism is crucial for the extraction of extracellular vesicles. A pure air environment maintains the stability of extracellular vesicles, prevents their structure and composition from being affected, and improves the extraction quality. Good filtration effects reduce the occurrence of equipment failures, extend the service life of the equipment, and reduce maintenance costs.

[0064] The setting of the filtration mechanism plays a key role in ensuring the extraction of extracellular vesicles. A pure air environment helps maintain the stability of extracellular vesicles and prevents impurities and harmful substances in the air from adversely affecting the structure and composition of extracellular vesicles, thereby improving the extraction quality of extracellular vesicles. Good filtration effects reduce the failure rate caused by air pollution inside the equipment, extend the service life of the equipment, and reduce maintenance costs.

[0065] In another embodiment of the present invention, a transmission mechanism for driving the valve-type filter 11 to flip is provided on the cleaning box 5, the base box 1 and the second rotating shaft 44, and the transmission mechanism includes: a first shaft 46 and a second shaft 48 rotatably installed on the cleaning box 5 and the base box 1 respectively; a third pulley 49 fixedly installed on the first shaft 46 and the second shaft 48 respectively, and the two third pulleys 49 are sleeved with the same second timing belt 50 for transmission; a fifth bevel tooth 51 fixedly installed on the first shaft 46; a third bevel tooth 45 fixedly installed on the second rotating shaft 44, and the third bevel tooth 45 can be engaged with and separated from the fifth bevel tooth 51, and is used to drive the valve-type filter 11 to flip; a fourth bevel tooth 47 fixedly installed on the first rotating shaft 39 and the second shaft 48 respectively, and the two fourth bevel teeth 47 are engaged with each other to drive the second shaft 48 to rotate.

[0066] In this embodiment, the valve-type filter disc 11 will flip over during cleaning, and the first rotating shaft 39 will rotate first, because the fourth bevel gear 47 on it meshes with the fourth bevel gear 47 on the second shaft 48, driving the second shaft 48 to rotate. The third pulley 49 on the second shaft 48 rotates, and the third pulley 49 on the first shaft 46 is also rotated through the second timing belt 50, thereby driving the first shaft 46 to rotate. The fifth bevel gear 51 on the first shaft 46 rotates accordingly, and when the filter housing 10 enters the cleaning box 5, the third bevel gear 45 meshes with the fifth bevel gear 51, and the fifth bevel gear 51 drives the third bevel gear 45 to rotate. Since the third bevel gear 45 is fixed on the second rotating shaft 44, the second rotating shaft 44 is connected to the valve-type filter disc 11, and finally the valve-type filter disc 11 is flipped.

[0067] Compared with the traditional driving method, the transmission mechanism of the present invention has significant advantages. The system composed of multiple shafts, pulleys, and bevel gears realizes efficient power transmission and flexible control, and the transmission is smooth, avoiding the valve-type filter 11 from turning over and getting stuck or inaccurate. The third bevel gear 45 and the fifth bevel gear 51 can be disengaged, and the operator can flexibly control the turning timing, thereby improving the convenience and flexibility of operation.

[0068] The transmission mechanism plays an important role in improving the cleaning work, and the stable flipping operation is helpful for backwashing the valve type filter 11.

[0069] In another embodiment of the present invention, a pin rod 57 is provided on the baffle 7, a first spring 58 is sleeved on the pin rod 57, a positioning block 59 for positioning is fixedly installed on the bottom end of the pin rod 57, a limiting block 60 is fixedly installed on one end of the slide plate 6, and two limiting grooves are provided on the slide plate 6, and both limiting grooves are adapted to the positioning block 59.

[0070] In this embodiment, when the baffle 7 is to be moved, the positioning block 59 at the bottom end of the pin rod 57 on the baffle 7 is initially located in the limiting groove at one end of the slide plate 6. The positioning block 59 needs to be pulled out by overcoming the elastic force of the first spring 58, and then the baffle 7 is pushed to slide on the slide plate 6. After reaching the appropriate position, it is released, and the positioning block 59 is embedded in another limiting groove of the slide plate 6 under the elastic force of the spring to achieve the positioning of the baffle 7. The limiting block 60 at one end of the slide plate 6 can prevent the positioning block 59 from sliding off, ensuring the stability of the structure; Compared with the traditional baffle positioning, the positioning structure of the present invention has obvious advantages. Through the cooperation of the pin 57, the first spring 58, the positioning block 59 and the limit groove, the baffle 7 can be accurately positioned, and the operator can fix it in different positions as needed to meet a variety of work scenarios. For example, when cleaning the filter housing 10, the baffle 7 can be placed in a suitable position, and after positioning, the baffle 7 is stable and does not slide. This positioning method is easy to operate, and only requires plugging and unplugging the positioning block 59, without the need for complex tools, which improves the convenience of operation; The positioning structure of the baffle 7 has a positive effect on the extracellular vesicle extraction device. Precise positioning ensures the stable movement of the filter housing 10, avoids collision damage caused by the instability of the baffle 7, and prolongs the life of the equipment. The convenient operation allows researchers to efficiently clean the filter housing 10, etc., and improve the efficiency of extracellular vesicle extraction.

[0071] In another embodiment of the present invention, solenoid valves 42 are provided on the first air duct 31 and the second air duct 34, a diverter plate 43 for diverting hot air is provided in the cleaning box 5, a mounting block is fixedly installed on the outer wall of the filter shell 10, and a limiting bolt that can be screwed to abut against the second rotating shaft 44 is provided on the mounting block. The limiting bolt can be screwed loose to separate it from the second rotating shaft 44 before cleaning the valve type filter 11.

[0072] In this embodiment, when the extracellular vesicle extraction device is in operation, the air volume of the first and second air ducts can be adjusted by controlling the solenoid valve 42 to accurately control the air flow interruption and flow rate. When cleaning the valve-type filter 11, the diverter plate 43 in the cleaning box 5 diverts the hot air to evenly dry and assist in cleaning the filter housing 10 and the valve-type filter 11. Before cleaning, loosen the limit bolts to allow the second rotating shaft 44 to rotate freely, which is convenient for all-round cleaning of the valve-type filter 11; after cleaning, tighten the bolts to fix its position to ensure filtering stability.

[0073] Compared with the traditional device, the new components of the present invention have significant advantages. The solenoid valve 42 accurately and efficiently controls the air duct, flexibly adjusts the air volume according to the experimental requirements and the equipment status, improves energy efficiency, and stabilizes the equipment temperature flow environment. The diverter plate 43 evenly distributes the hot air, enhances the cleaning and drying effect, and prolongs the life of the components. The limit bolt facilitates the cleaning and maintenance of the valve type filter 11, avoids incomplete cleaning, and ensures working stability and filtering accuracy.

[0074] The coordinated design of the solenoid valve 42, the diverter plate 43 and the limit bolt promotes the extraction of extracellular vesicles. Precise air duct control and hot air diversion keep the equipment in the best condition and improve the extraction quality. Convenient cleaning and maintenance improve equipment reliability, reduce experimental interruptions, improve experimental efficiency, and enhance device stability and repeatability.

[0075] In summary, compared with the relevant technologies, through standardized sample preparation, centrifugation, filtration, washing and re-centrifugation steps, the cell exosomes are gradually purified to obtain high-quality products. The laterally adjustable filter shell 10 cooperates with the washing box 5 and the adjustment mechanism to facilitate cleaning, simplify the process, reduce operating losses, and improve extraction efficiency. The cooling mechanism in the base box 1 and the centrifugal box 2 solves the problem of high-speed rotation heat generation, ensures the cell exosome extraction environment, improves quality and experimental reliability, and extends the life of the equipment. The placement seat 12, beaker 13 and the liquid outlet pipe 65 of the filter shell 10 on the top of the base box 1 are precisely docked to achieve accurate collection of filtrate, improve efficiency and basic quality of subsequent experiments. The drain pipe 52 of the washing box 5 and the collection box 53 in the base box 1 effectively collect dirty liquid, keep the device clean, and ensure the cleaning effect and extraction quality. The exhaust pipe 55 and the drain pipe 56 of the centrifugal box 2 exhaust heat and drain water to avoid high-temperature water accumulation in the box, and ensure stable operation of the equipment and extraction quality.

[0076] The present invention also provides an extraction method of an exosome extraction device, and the extraction method comprises the following steps: S1: Sample preparation: Select liquid of cell solution, perform biological culture, and obtain sample solution. Take a certain amount of sample solution, add it into the centrifuge test tube 61, place the centrifuge test tube 61 in the placement groove of the centrifuge seat 3, and clamp the centrifuge test tube 61 through the clamp block 63 under the action of the second spring 62; S2: First centrifugation: Close the cover 54 on the top of the centrifuge box 2, set the speed and centrifugation time through the control module, start the centrifuge seat 3 to rotate, and perform the first centrifugation operation on the sample solution. During this process, the Peltier refrigeration sheet 4 on the inner wall of the centrifuge box 2 works to cool the high-speed rotating centrifuge seat 3 and the centrifuge test tube 61; at the same time, the fan blades 33 of the air supply box 16 in the base box 1 rotate, and air is supplied to the centrifuge box 2 through the first air duct 31 for cooling, the exhaust pipe 55 exhausts and dissipates heat, and the drain pipe 56 discharges condensed water; S3: First filtration: After the first centrifugation is completed, the liquid after centrifugation in the centrifuge test tube 61 is poured into the filter shell 10, and the liquid is filtered through the valve type filter 11. The supernatant after filtration enters the beaker 13 through the liquid outlet pipe 65 to remove the sediment, collect the supernatant, and remove Living cells and larger impurities; S4: Cleaning operation: Use the adjustment mechanism to move the filter shell 10 into the cleaning box 5, and the flushing pipe 14 sprays the cleaning liquid to rinse the filter shell 10 and the valve-type filter disc 11. The cleaned dirty liquid is discharged into the collection box 53 through the drain pipe 52; S5: Second centrifugation: The supernatant in the beaker 13 is added to the centrifuge tube 61 again, and the centrifugation time is set again through the control module to perform a second centrifugation operation. During the centrifugation process, the cooling components of the Peltier cooling plate 4 and the air supply box 16 work synchronously; S6: Second filtration: After the second centrifugation is completed, the first filtration operation is repeated to obtain the supernatant again, and this time the dead cells and cell fragments are removed; S7: Third centrifugation and exosome preparation: The supernatant obtained by the second filtration is added to the centrifuge tube 61 again, and the centrifugation time is set again through the control module for strong centrifugation. Then, the liquid in the centrifuge tube 61 is poured into the filter shell 10 for filtration. This time, the supernatant is removed to obtain a precipitate. The precipitate was resuspended in 100 μL PBS for purification and extraction to obtain cell exosomes.

[0077] In the several embodiments provided in this application, it should be understood that the disclosed device can be implemented in other ways.

[0078] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the scope of protection of the invention. Obviously, the described embodiments are only some embodiments of the present invention, rather than all embodiments. Based on these embodiments, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in this field can still combine, add, delete or make other adjustments to the features in the various embodiments of the present invention according to the circumstances without conflict, without making creative work, so as to obtain different other technical solutions that do not deviate from the concept of the present invention in essence, and these technical solutions also belong to the scope of protection of the present invention.

Claims

1. A device for extracting extracellular vesicles, characterized in that: include: A base box, wherein a centrifuge box is disposed on the top of the base box, and a rotatable centrifuge seat is disposed in the centrifuge box, and the centrifuge seat is used to place a centrifuge test tube containing a cell sample solution; A filter housing is arranged above the base box and located at the side of the centrifuge box, the filter housing can move laterally, and a rotatably adjustable valve-type filter is arranged in the filter housing, the valve-type filter is used to filter and process the cell sample solution after centrifugation; A cleaning box is mounted on the top of the base box. The cleaning box is located on one side of the lateral movement track of the filter shell and has an inlet and outlet for the filter shell to enter and exit. A flushing pipe is fixedly installed in the cleaning box for spraying cleaning liquid to flush the filter shell and the valve-type filter disc located below after the filter shell enters the cleaning box; A third air duct fixedly connected to the cleaning box, used for introducing hot air to dry the filter housing and the valve-type filter disc; An adjusting mechanism disposed on the cleaning box for adjusting the lateral movement of the filter housing; and The cooling mechanism arranged in the base box and the centrifugal box is used for supplying air to cool the inside of the centrifugal box.

2. The device for extracting extracellular vesicles according to claim 1, characterized in that: The regulating mechanism comprises: A slide plate is fixed on the top of the cleaning box, and a baffle is slidably sleeved on the slide plate to cover the inlet and outlet of the filter shell after the filter shell enters the cleaning box. A connecting frame is fixedly mounted on the baffle, and a placement frame for placing the filter shell is fixedly mounted on the connecting frame. The connecting frame and the placement frame with the filter shell placed thereon can both follow the movement of the baffle to enter and exit along the inlet and outlet of the cleaning box.

3. The device for extracting extracellular vesicles according to claim 1, characterized in that: The cooling mechanism comprises: A plurality of Peltier cooling sheets mounted on the inner wall of the centrifugal box for cooling; An air supply box fixed in the base box, wherein a mounting rod is provided in the air supply box, and a fan blade is mounted on the mounting rod; Rotate a transmission rod installed on the air supply box, one end of the transmission rod being fixedly connected to the installation rod via a coupling; A first air duct is fixedly connected to the air supply box, and an air outlet end of the first air duct is connected to the centrifugal box.

4. The exosome extraction device according to claim 1, characterized in that: A placement seat located between the centrifuge box and the cleaning box is provided on the top of the base box, and at least two beakers are provided on the placement seat. The beakers are placed on the movement trajectory of the filter shell so that the beakers can receive the solution filtered by the filter shell. The filter shell is fixedly connected with a liquid outlet pipe, and the liquid outlet pipe is used to guide the supernatant filtered in the filter shell to the corresponding beaker.

5. The device for extracting extracellular vesicles according to claim 2, characterized in that: The bottom of the cleaning box is fixedly connected with a drain pipe for discharging dirty liquid, and the liquid outlet end of the drain pipe extends into the base box. A collecting box is arranged in the base box, and the collecting box is used to receive and collect the dirty liquid discharged from the drain pipe.

6. The device for extracting extracellular vesicles according to claim 2, characterized in that: An exhaust pipe and a drain pipe are fixedly connected to one side of the centrifugal box. The exhaust pipe is used for exhausting and dissipating heat, and the drain pipe is used for discharging condensed water.

7. The device for extracting extracellular vesicles according to claim 1, characterized in that: A detachable cover is provided on the top of the centrifuge box, a handle for grasping is fixedly mounted on the cover, and a liquid supply pipe is fixedly mounted on the liquid inlet end of the flushing pipe, and the liquid supply pipe is used to connect to a cleaning liquid supply device.

8. The device for extracting extracellular vesicles according to claim 1, characterized in that: The centrifugal seat is provided with a placement groove and a plurality of air holes, a second spring is symmetrically fixedly installed on the inner wall of the placement groove, and a clamping block for clamping the centrifugal test tube is fixedly installed on the second spring.

9. The device for extracting extracellular vesicles according to claim 2, characterized in that: A limiting ring is fixedly sleeved on the filter shell, a plug-in block is fixedly installed on the bottom of the limiting ring, a groove adapted to the plug-in block is provided on the placement rack, a second rotating shaft is rotatably installed on the filter shell through a bearing, and the second rotating shaft is fixedly connected to the valve-type filter disc.

10. A method for extracting exosomes using the exosome extraction device according to any one of claims 1 to 9, characterized in that: The following steps are involved: S1: Sample preparation: Select liquid of cell solution, perform biological culture, and obtain sample solution. Take a certain amount of sample solution, add it into a centrifuge test tube, place the centrifuge test tube in the placement slot of the centrifuge seat, and clamp the centrifuge test tube with a clamp block under the action of the second spring; S2: First centrifugation: Close the cover on the top of the centrifuge box, set the speed and centrifugation time through the control module, start the centrifuge seat to rotate, and perform the first centrifugation operation on the sample solution. During this process, the Peltier cooling sheet on the inner wall of the centrifuge box works to cool the high-speed rotating centrifuge seat and centrifuge test tube; at the same time, the fan blades of the air supply box in the base box rotate, supply air to the centrifuge box through the first air duct for cooling, exhaust pipe exhausts heat, and the drain pipe discharges condensed water; S3: First filtration: After the first centrifugation is completed, the liquid in the centrifuge tube is poured into the filter housing, and the liquid is filtered through the valve filter. The filtered supernatant enters the beaker through the outlet pipe to remove the sediment, collect the supernatant, and remove the living cells and larger impurities; S4: Cleaning operation: Use the adjustment mechanism to move the filter housing into the cleaning box, and use the flushing pipe to spray the cleaning liquid to flush the filter housing and the valve filter disc. The cleaned dirty liquid is discharged into the collection box through the drain pipe; S5: Second centrifugation: the supernatant in the beaker is added to the centrifuge tube again, the centrifugation time is re-set through the control module, and the second centrifugation operation is performed. During the centrifugation process, the cooling components of the Peltier cooling sheet and the air supply box work synchronously; S6: Second filtration: After the second centrifugation, the first filtration operation is repeated to obtain the supernatant again, and this time the dead cells and cell debris are removed; S7: Third centrifugation and exosome preparation: The supernatant obtained from the second filtration was added to the centrifuge tube again, and the centrifugation time was set again through the control module for strong centrifugation. Then the liquid in the centrifuge tube was poured into the filter shell for filtration. The supernatant was removed this time to obtain a precipitate. The precipitate was resuspended in μL PBS for purification and extraction to obtain cell exosomes.

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