Extracellular vesicle separation device

Abstract

The utility model relates to the technical field of cell separation, in particular to an extracellular vesicle separation device, which comprises a support, an inlet assembly, a filter assembly, a separation chip assembly, a collection assembly and a vacuum assembly, the collection assembly is arranged on the support, the separation chip assembly is arranged on the collection assembly, and the vacuum assembly is arranged on the collection assembly. The filter assembly is arranged on the separation chip assembly, the vacuum assembly is arranged on the support, the inlet assembly comprises a feeding pipe, a valve, a sealing cover and a sealing ring, the sealing cover is arranged on the filter assembly, the feeding pipe is arranged on the sealing cover, the valve is arranged on the feeding pipe, and the sealing ring is arranged on the sealing cover. The whole machine can be sealed through the inlet assembly, the interior of the machine is in a vacuum state through the vacuum assembly, it is guaranteed that the experiment process is conducted in the vacuum state, time consumed in the whole separation process is shortened, and the whole sample treatment speed is increased.

Description

Technical Field

[0001] The present application relates to the field of cell separation technology, and in particular to an extracellular vesicle separation device. Background Art

[0002] Extracellular vesicles are a class of nano- to micro-sized membrane vesicles secreted by cells, which play an important role in intercellular communication. Extracellular vesicles include different types such as exosomes, microvesicles and apoptotic bodies. Due to their potential applications in disease diagnosis, treatment and biomarker discovery, the separation and purification of extracellular vesicles has become an important research field. Extracellular vesicle separation devices and methods include ultracentrifugation, size exclusion chromatography, immunoaffinity chromatography, precipitation, microfluidics, and field flow fractionation.

[0003] At present, the Chinese utility model patent application with the publication date of January 31, 2023 and the publication number of CN 218404184U provides an extracellular vesicle separation device, including a filtration separation chamber and a collection chamber, wherein a plurality of filter membranes are vertically spaced in the filtration separation chamber, and the two sides of the filter membranes are connected to the side walls of the filtration separation chamber, and a baffle is hinged at the bottom of each filter membrane; an inlet and a liquid inlet are provided at the top of the filtration separation chamber, and the inlet and the liquid inlet are connected by a control valve, and a sample outlet is provided at the lower part of the side of the filtration separation chamber away from the inlet; the filtration separation chamber is separated from the collection chamber by a slide, and the slide slide is slidably arranged in the chamber, wherein the height of the baffle is the same as the thickness of the slide, and the collection chamber includes a plurality of collection chambers, and the spacing between the plurality of collection chambers and the bottom of the plurality of filter membranes corresponds one to one. The present application has a multi-stage filtration and separation function, and can perform multi-stage filtration and separation on the extracellular vesicles, and rinse and clean them to obtain substances of different particle sizes, with good separation effect and high separation efficiency.

[0004] The extracellular vesicle separation device in the related art uses multiple filter membranes to filter the sample, thereby collecting substances of different particle sizes. However, the overall flow rate of this method is slow, which causes the entire separation process to take a long time, limiting the sample processing speed. Utility Model Content

[0005] An embodiment of the present application provides an extracellular vesicle separation device to improve the technical problem in the related art that the overall flow rate is slow, resulting in a long separation process and limiting the sample processing speed.

[0006] An embodiment of the present application provides an extracellular vesicle separation device, comprising a bracket, an inlet component, a filtering component, a separation chip component, a collection component and a vacuum component, wherein the collection component is arranged on the bracket, the separation chip component is arranged on the collection component, the filtering component is arranged on the separation chip component, and the vacuum component is arranged on the bracket, the inlet component comprises a feed pipe, a valve, a sealing cover and a sealing ring, the sealing cover is arranged on the filtering component, the feed pipe is arranged on the sealing cover, the valve is arranged on the feed pipe, and the sealing ring is arranged on an end of the sealing cover away from the feed pipe.

[0007] The above-mentioned technical scheme in the embodiment of the present application has at least the following technical effects: before injecting the extracellular vesicles, the vacuum component is turned on to put the inside of the machine in a vacuum state. During the separation of the extracellular vesicles, the operator uses the injection pump to inject the sample liquid from the feed tube through the electric valve. The sealing cover and the sealing ring ensure the sealing effect of the entire machine, reducing the probability of reducing the separation efficiency due to leakage.

[0008] The extracellular vesicle separation device provided in the embodiment of the present application can seal the entire machine through an inlet component, and put the inside of the machine in a vacuum state through a vacuum component, thereby ensuring that the experimental process is carried out in a vacuum state, reducing the time consumption of the entire separation process, and improving the speed of overall sample processing.

[0009] In some embodiments, the valve includes a fixed plate, a blocking plate, a frustum, a spring and a baffle. The fixed plate is arranged on the feed pipe and a circular hole is formed on the fixed plate. The baffle is arranged on the feed pipe and a through hole is formed on the baffle. One end of the spring abuts against the baffle. A blocking plate is arranged on the other end of the spring and a plurality of connecting holes are formed on the blocking plate. A frustum is arranged on the end of the blocking plate away from the spring and the frustum abuts against the fixed plate through the circular hole.

[0010] In some embodiments, the collection assembly includes a collection box and a liquid outlet pipe, the collection box is disposed on a bracket, and the liquid outlet pipe is disposed on the collection box.

[0011] In some embodiments, the vacuum assembly includes a vacuum pump and an air suction pipe, the vacuum pump is disposed on a bracket, one end of the air suction pipe is disposed on the vacuum pump, and the other end of the air suction pipe is disposed on a collection box.

[0012] In some embodiments, the separation chip assembly includes a separation chamber and a microfluidic chip. The separation chamber is disposed on an end of the liquid outlet tube away from the collection box, and the microfluidic chip is disposed in the separation chamber.

[0013] In some embodiments, the filtration assembly includes an injector, a first PMMA ring, a second PMMA ring, an AAO membrane and a connecting tube, wherein the connecting tube is arranged at one end of the separation chamber away from the liquid outlet pipe, the injector is arranged at one end of the connecting tube away from the separation chamber, the first PMMA ring is arranged in the injector, the second PMMA ring is arranged in the injector, one end of the AAO membrane abuts against the first PMMA ring, the other end of the AAO membrane abuts against the second PMMA ring, and the sealing cover is arranged on one end of the injector away from the connecting tube.

[0014] In some embodiments, a handle is provided on the bracket, a display screen is provided on the bracket, and a button is provided on the bracket. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 A schematic diagram of the three-dimensional structure of the extracellular vesicle separation device provided in an embodiment of the present application;

[0016] Figure 2 A schematic diagram of the cross-sectional structure of an extracellular vesicle separation device provided in an embodiment of the present application;

[0017] Figure 3 for Figure 2 Enlarged view of point A in the middle;

[0018] Figure 4 A schematic diagram of the explosion structure of a valve provided in an embodiment of the present application;

[0019] Among them, the reference numerals in the figure are:

[0020] 1. Bracket; 2. Inlet assembly; 21. Feed pipe; 22. Valve; 221. Fixing plate; 222. Round hole; 223. Blocking plate; 224. Cone; 225. Connecting hole; 226. Spring; 227. Baffle; 228. Through hole; 23. Sealing cover; 24. Sealing ring; 3. Filter assembly; 31. Sampler; 32. First PMMA ring; 33. Second PMMA ring; 34. AAO membrane; 35. Connecting pipe; 4. Separation chip assembly; 41. Separation chamber; 42. Microfluidic chip; 5. Collection assembly; 51. Collection box; 52. Liquid outlet pipe; 6. Vacuum assembly; 61. Vacuum pump; 62. Suction pipe; 7. Handle; 8. Button; 9. Display screen. DETAILED DESCRIPTION

[0021] Based on this, in order to improve the technical problem in the related art that the overall flow rate is slow, resulting in a long separation process and limiting the sample processing speed, the embodiments of the present application provide the following solution.

[0022] Please also read Figures 1 to 4An embodiment of the present application provides an extracellular vesicle separation device, which includes a bracket 1, an inlet component 2, a filtering component 3, a separation chip component 4, a collecting component 5 and a vacuum component 6. The collecting component 5 is arranged on the bracket 1, the separation chip component 4 is arranged on the collecting component 5, the filtering component 3 is arranged on the separation chip component 4, the vacuum component 6 is arranged on the bracket 1, and the inlet component 2 is arranged on the filtering component 3.

[0023] In some embodiments, please refer to Figures 1 to 3 The collecting assembly 5 includes a collecting box 51 and a liquid outlet pipe 52. The collecting box 51 is arranged on the support 1, and the liquid outlet pipe 52 is arranged on the collecting box 51. The vacuum assembly 6 includes a vacuum pump 61 and an air suction pipe 62. The vacuum pump 61 is arranged on the support 1, one end of the air suction pipe 62 is arranged on the vacuum pump 61, and the other end of the air suction pipe 62 is arranged on the collecting box 51. The separation chip assembly 4 includes a separation chamber 41 and a microfluidic chip 42. The separation chamber 41 is arranged on one end of the liquid outlet pipe 52 away from the collecting box 51, and the microfluidic chip 42 is arranged in the separation chamber 41. The filter assembly 3 includes an injector 31, a first PMMA ring 32, a second PMMA ring 33, an AAO membrane 34 and a connecting tube 35. The connecting tube 35 is arranged on the end of the separation chamber 41 away from the liquid outlet pipe 52. The injector 31 is arranged on the end of the connecting tube 35 away from the separation chamber 41. The first PMMA ring 32 is arranged in the injector 31, and the second PMMA ring 33 is arranged in the injector 31. One end of the AAO membrane 34 abuts on the first PMMA ring 32, and the other end of the AAO membrane 34 abuts on the second PMMA ring 33.

[0024] With such a configuration, before the user performs the separation operation, the vacuum pump 61 is turned on to put the entire machine in a vacuum state. After the operator injects the sample liquid into the sampler 31, the sample liquid is filtered through the AAO membrane 34, and then reaches the separation chamber 41 through the connecting tube 35, and then the extracellular vesicles are separated through the microfluidic chip 42. The separated extracellular vesicles then enter the collection box 51. The vacuum state can accelerate the filtration process of the sample liquid through the AAO membrane 34, thereby improving the separation efficiency. The negative pressure under the vacuum state can promote the rapid passage of the liquid through the filter membrane and reduce the separation time. Under the vacuum state, the sample liquid can be more evenly distributed on the filter membrane, reducing the sample residue in the pipeline, thereby reducing the sample loss. The microfluidic chip 42 technology can accurately control the flow path and speed of the sample liquid in the chip, thereby improving the separation purity of the extracellular vesicles. By accurately controlling the flow rate and flow path, the extracellular vesicles of the target size can be separated more effectively. The closed system under the vacuum state can reduce the entry of external pollutants, thereby reducing the risk of sample contamination. The separation system using the vacuum pump 61 and the microfluidic chip 42 can more easily realize automated operation, reduce errors caused by human operation, and improve the repeatability and reliability of the experiment. The automated and efficient separation process can save the time and labor of the experimenters, allowing them to focus on other experimental steps or data analysis.

[0025] In some embodiments, please refer to Figures 1 to 4 The inlet assembly 2 includes a feed pipe 21, a valve 22, a sealing cover 23 and a sealing ring 24. The sealing cover 23 is arranged on the end of the sampler 31 away from the connecting pipe 35. The feed pipe 21 is arranged on the sealing cover 23. The valve 22 is arranged on the feed pipe 21. The sealing ring 24 is arranged on the end of the sealing cover 23 away from the feed pipe 21. The valve 22 includes a fixing plate 221, a blocking plate 223, a round table 224, a spring 226 and a baffle 227. The fixing plate 221 is arranged on On the feed pipe 21, a circular hole 222 is provided on the fixed plate 221, the baffle 227 is arranged on the feed pipe 21, a through hole 228 is provided on the baffle 227, one end of the spring 226 abuts on the baffle 227, a blocking plate 223 is provided on the other end of the spring 226, a plurality of connecting holes 225 are provided on the blocking plate 223, a frustum 224 is provided on the end of the blocking plate 223 away from the spring 226, and the frustum 224 abuts against the fixed plate 221 through the circular hole 222.

[0026] With such a configuration, during the separation of extracellular vesicles, the operator injects the sample liquid from the feed tube 21 through the valve 22 through the syringe pump. The syringe pump first abuts against the fixed plate 221, and the syringe pump compresses the spring 226 downward by abutting against the truncated cone 224. At the same time, the spring 226 drives the blocking plate 223 and the truncated cone 224 to move downward at the same time. The sample liquid flows down through the connecting hole 225 on the blocking plate 223, and then flows into the sample injector 31 through the through hole 228 on the baffle 227. After the sample liquid is injected, the syringe pump is taken out, and the blocking plate 223 and the truncated cone 224 rebound under the action of the spring 226. The truncated cone 224 seals the fixed plate 221 through the circular hole 222 on the fixed plate 221 to ensure the vacuum state in the machine. In this way, the syringe pump can accurately control the injection amount of the sample liquid to ensure the consistency and repeatability of each experiment, which is crucial for the separation and subsequent analysis of extracellular vesicles. Through the automated operation of the mechanical device, the error of human operation can be reduced, and the accuracy and efficiency of the experiment can be improved. After the sample liquid is added, the mechanical device can automatically seal the circular hole 222 on the fixed plate 221 to maintain the vacuum state in the machine. The vacuum state is very important for the separation of extracellular vesicles because it can reduce the oxidation and contamination of the sample, and at the same time help stabilize and preserve the sample. By using components such as the sealing plate 223 and the round table 224, the sample liquid can be effectively prevented from leaking or contaminating during the addition process, ensuring the cleanliness and accuracy of the experiment. This design is usually compact in structure, occupies a small space, and is suitable for laboratory environments.

[0027] In some embodiments, please refer to Figure 1 to Figure 2 The bracket 1 is provided with a handle 7, the bracket 1 is provided with a display screen 9, and the bracket 1 is provided with a button 8.

[0028] With such a configuration, the design of the handle 7 makes the device easy to move and position, and it is convenient for the experimenter to adjust the position of the device as needed, especially when the laboratory space is limited. The display screen 9 can provide real-time operation information and feedback, such as the current operation step, the status of the separation process, the remaining time, etc. This helps the experimenter to monitor the progress of the experiment, find and solve problems in time, and the setting of the button 8 allows the user to directly interact with the device to perform basic operations such as start, stop, pause, etc. This intuitive control method reduces the learning curve, so that new users can also get started quickly. Through the combination of the display screen 9 and the button 8, the user can quickly set and adjust the experimental parameters, reducing the time of finding and adjusting the settings, thereby improving the experimental efficiency. The intuitive user interface and clear button 8 identification can reduce operating errors. In an emergency, the button 8 can quickly stop the operation of the equipment to prevent accidents or reduce the impact of accidents. Some advanced display screens 9 may also have a data recording function, which can record experimental data for subsequent analysis and recording.

[0029] The implementation principle of an extracellular vesicle separation device in an embodiment of the present application is as follows: before performing the separation operation, the user turns on the vacuum pump 61 to put the entire machine in a vacuum state. During the separation of the extracellular vesicles, the operator uses the syringe pump to inject the sample liquid from the feed tube 21 through the valve 22. The syringe pump first abuts against the fixed plate 221, and the syringe pump compresses the spring 226 downward by abutting the round table 224. At the same time, the spring 226 drives the blocking plate 223 and the round table 224 to move downward at the same time. The sample liquid flows down through the connecting hole 225 on the blocking plate 223, and then flows into the sample injector 31 through the through hole 228 on the baffle 227. After the sample liquid is added, the injection pump is taken out, and the sealing plate 223 and the round table 224 rebound under the action of the spring 226. The round table 224 seals the fixed plate 221 through the circular hole 222 on the fixed plate 221 to ensure the vacuum state in the machine. The sealing cover 23 and the sealing ring 24 ensure the sealing effect of the entire machine to reduce the probability of reducing the separation efficiency due to leakage. After the operator injects the sample liquid into the sampler 31, the sample liquid is filtered through the AAO membrane 34, and then reaches the separation chamber 41 through the connecting tube 35, and then the extracellular vesicles are separated through the microfluidic chip 42, and then the separated extracellular vesicles enter the collection box 51.

[0030] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. An extracellular vesicle separation device, characterized in that: The invention comprises a support (1), an inlet assembly (2), a filter assembly (3), a separation chip assembly (4), a collection assembly (5) and a vacuum assembly (6); the collection assembly (5) is arranged on the support (1); the separation chip assembly (4) is arranged on the collection assembly (5); the filter assembly (3) is arranged on the separation chip assembly (4); the vacuum assembly (6) is arranged on the support (1); the inlet assembly (2) comprises a feed pipe (21), a valve (22), a sealing cover (23) and a sealing ring (24); the sealing cover (23) is arranged on the filter assembly (3); the feed pipe (21) is arranged on the sealing cover (23); the valve (22) is arranged on the feed pipe (21); and the sealing ring (24) is arranged on an end of the sealing cover (23) away from the feed pipe (21).

2. The extracellular vesicle separation device according to claim 1, characterized in that: The valve (22) comprises a fixed plate (221), a blocking plate (223), a round platform (224), a spring (226) and a baffle (227); the fixed plate (221) is arranged on the feed pipe (21); a round hole (222) is provided on the fixed plate (221); the baffle (227) is arranged on the feed pipe (21); a through hole (228) is provided on the baffle (227); one end of the spring (226) abuts against the baffle (227); the other end of the spring (226) is provided with a blocking plate (223); a plurality of connecting holes (225) are provided on the blocking plate (223); a round platform (224) is provided at one end of the blocking plate (223) away from the spring (226); the round platform (224) abuts against the fixed plate (221) through the round hole (222).

3. The extracellular vesicle separation device according to claim 2, characterized in that: The collecting assembly (5) comprises a collecting box (51) and a liquid outlet pipe (52); the collecting box (51) is arranged on the bracket (1), and the liquid outlet pipe (52) is arranged on the collecting box (51).

4. The extracellular vesicle separation device according to claim 3, characterized in that: The vacuum assembly (6) comprises a vacuum pump (61) and an air suction pipe (62); the vacuum pump (61) is arranged on the bracket (1); one end of the air suction pipe (62) is arranged on the vacuum pump (61); and the other end of the air suction pipe (62) is arranged on the collection box (51).

5. The extracellular vesicle separation device according to claim 4, characterized in that: The separation chip assembly (4) comprises a separation chamber (41) and a microfluidic chip (42); the separation chamber (41) is arranged on one end of the liquid outlet tube (52) away from the collection box (51); and the microfluidic chip (42) is arranged in the separation chamber (41).

6. The extracellular vesicle separation device according to claim 5, characterized in that: The filter assembly (3) comprises a sample injector (31), a first PMMA ring (32), a second PMMA ring (33), an AAO membrane (34) and a connecting tube (35); the connecting tube (35) is arranged on one end of the separation chamber (41) away from the liquid outlet tube (52); the sample injector (31) is arranged on one end of the connecting tube (35) away from the separation chamber (41); the first PMMA ring (32) is arranged in the sample injector (31); the second PMMA ring (33) is arranged in the sample injector (31); one end of the AAO membrane (34) abuts against the first PMMA ring (32); the other end of the AAO membrane (34) abuts against the second PMMA ring (33); and the sealing cover (23) is arranged on one end of the sample injector (31) away from the connecting tube (35).

7. The extracellular vesicle separation device according to claim 6, characterized in that: The bracket (1) is provided with a handle (7), the bracket (1) is provided with a display screen (9), and the bracket (1) is provided with a button (8).

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