Experimental device for extracting exosome

Abstract

The invention discloses an experimental device for extracting exosomes, and belongs to the field of biological test equipment. The device comprises a centrifugal machine, a separation mechanism is installed in the centrifugal machine, the separation mechanism comprises a first cylinder, a second cylinder connected to the first cylinder and a third cylinder connected to the second cylinder, a primary sample is placed in the first cylinder, after primary centrifugation, the sample in the first cylinder enters the second cylinder, after secondary centrifugation, the sample in the second cylinder enters the third cylinder, and the sample in the third cylinder enters the third cylinder. The sample in the second cylinder enters the third cylinder; after the device is shut down, the centrifugal cylinder does not need to be replaced for multiple times, multiple centrifugal separation can be achieved only by transferring samples in the three cylinder bodies, and the operation efficiency is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological experimental equipment, in particular to an experimental device for extracting exosomes. BACKGROUND

[0002] Myocardial exosomes are important vesicular structures secreted by myocardial cells, and the extraction purity and efficiency thereof directly affect subsequent functional research and application development. Centrifugal separation is a core and key step in the current myocardial exosome extraction process. The existing myocardial exosome sample pretreatment stage needs to be subjected to multiple gradient centrifugations to gradually remove impurities such as intact cells, cell fragments and large-size vesicles. In the traditional operation, after each centrifugation is completed, the sample needs to be transferred from the current centrifuge tube to another pre-cooled centrifuge tube, and then re-installed in the centrifuge to set the parameters for the next centrifugation. This process not only needs to frequently change the centrifuge tube and repeatedly clamp and position, but also is complicated and time-consuming, thereby reducing the overall extraction efficiency. Meanwhile, during the multiple sample transfer processes, the exosome membrane structure is easily damaged due to improper operation, or the sample is easily contaminated due to exposure to the external environment, thereby affecting the subsequent extraction purity and exosome activity. To solve the above problems, there is an urgent need for an experimental device capable of simplifying the multiple centrifugation operation process and reducing the number of centrifuge tube changes, so as to improve the efficiency and reliability of myocardial exosome extraction. SUMMARY

[0003] The present application aims to provide an experimental device for extracting exosomes, which solves the problem of multiple sample transfers after centrifugation of the existing experimental device.

[0004] To achieve the above-mentioned purpose, the present application provides the following technical scheme: an experimental device for extracting exosomes, comprising a centrifuge, wherein a separation mechanism is installed in the centrifuge, the separation mechanism comprises a first cylinder, a second cylinder connected to the first cylinder, and a third cylinder connected to the second cylinder, a primary sample is placed in the first cylinder, after primary centrifugation, the sample in the first cylinder enters the second cylinder, and after secondary centrifugation, the sample in the second cylinder enters the third cylinder.

[0005] Preferably, an inner thread is formed in the port of the first cylinder, an outer thread matched with the inner thread is arranged at the bottom of the second cylinder, a shaft body is connected through the middle parts of the first cylinder, the second cylinder and the third cylinder, a first piston plate is slidably connected in the interior of the first cylinder, the first piston plate is connected to the bottom end of the shaft body, a second piston plate is slidably connected in the second cylinder, an elastic pipeline is in communication between the first piston plate and the second piston plate, and a one-way valve is arranged on the elastic pipeline. The second piston plate is provided with a through hole matched with the shaft body, the first piston plate is fixedly connected with the bottom end of the shaft body, and the sample in the first cylinder body enters the second cylinder body through the elastic pipeline when the shaft body moves downward.

[0006] Preferably, an annular frame is internally threadedly connected with the top port of the second cylinder body and externally threadedly connected with the third cylinder body, a filter membrane is installed on the annular frame, and the sample after the second centrifugation can pass through the filter membrane and enter the third cylinder body when the second piston plate moves upward.

[0007] Preferably, a wedge block is radially and slidingly connected with the middle part of the shaft body, a spring is fixedly connected between the wedge block and the shaft body, the wedge block can pass through the second piston plate after the shaft body moves downward, and the shaft body can drive the second piston plate to move upward after the wedge block passes through the second piston plate.

[0008] Preferably, the side wall of the first cylinder body close to the second cylinder body, the side wall of the second cylinder body close to the third cylinder body and the top wall of the third cylinder body are all provided with a vent hole.

[0009] Preferably, a machine cover is hingedly connected with the centrifugal machine, a circular hole coaxial with the third cylinder body is formed in the machine cover, a plurality of electric telescopic rods are radially and slidingly connected in the circular hole, L-shaped plates are fixedly connected with the ends of the electric telescopic rods, two ball bearings are installed on the inner walls of the L-shaped plates, and the two ball bearings roll on the top wall and the side wall of the third cylinder body after the electric telescopic rods are elongated.

[0010] Preferably, a lock block is rotatably connected with the machine cover, a lock groove matched with the lock block is arranged on the centrifugal machine, and the lock block can be clamped in the lock groove after rotation.

[0011] Preferably, a pressure-sensitive switch is arranged in the lock groove, and the lock block can press the pressure-sensitive switch when clamped in the lock groove, so that the electric telescopic rods are elongated.

[0012] Preferably, a pneumatic cylinder is fixedly connected with the machine cover, an installation frame is fixedly connected with the output end of the pneumatic cylinder, and a clamping block is installed on the installation frame. A limiting ring is rotatably connected with the upper part of the shaft body, and a tapered block is rotatably connected with the top of the shaft body, the clamping block can be clamped between the tapered block and the limiting ring when the machine cover is closed, and the pneumatic cylinder can drive the shaft body to move axially through extension and contraction.

[0013] Preferably, the clamping block is elastically and slidingly connected with the installation frame.

[0014] Compared with the prior art, the centrifugal machine has the following beneficial effects: The application synchronously rotates three barrels by starting the centrifuge, so that the sample is centrifuged, and the un-adherent complete myocardial cells and large cell groups are removed. After the first centrifugation is completed, the operation of the centrifuge is stopped, the supernatant in the first barrel is transferred to the second barrel, and then the centrifugation is performed again to remove the broken cell fragments, mitochondria and other large-particle impurities. After the centrifugation is completed, the supernatant in the second barrel is filtered into the third barrel to remove the microvesicles and protein aggregates with a diameter greater than 220 nanometers. Thus, the sample pretreatment is completed. After the device is stopped, multiple centrifugation separations can be achieved by transferring the sample in the three barrels, and the operation efficiency is improved. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 It is a schematic diagram of the overall structure of the application; Figure 2 It is a schematic diagram of the structure of the cover of the application; Figure 3 It is a schematic diagram of the structure of the card block of the application; Figure 4 It is a schematic diagram of the structure of the ball of the application; Figure 5 It is a schematic diagram of the structure of the three barrels in the initial state of the application; Figure 6 It is a schematic diagram of the structure of the sample in the first barrel of the application after being transferred to the second barrel; Figure 7 It is a schematic diagram of the structure of the sample in the second barrel of the application after being transferred to the third barrel; Figure 8 It is a schematic diagram of the structure of the shaft body of the application; Figure 9 It is a schematic diagram of the structure of the application Figure 8 A part of the structure of the application.

[0016] In the figure: 100, centrifuge; 110, control panel; 120, cover; 130, lock block; 140, lock groove; 150, N-shaped frame; 160, air cylinder; 170, mounting frame; 180, card block; 190, electric telescopic rod; 191, L-shaped plate; 192, ball; 200, first barrel; 210, first piston plate; 220, elastic pipeline; 300, second barrel; 310, second piston plate; 320, wedge block; 330, spring; 340, ring frame; 350, filter membrane; 400, third barrel; 410, shaft body; 420, limiting ring; 430, taper block. DETAILED DESCRIPTION

[0017] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0018] Reference Figures 1-9 This embodiment provides a technical solution: an experimental device for extracting exosomes, including a centrifuge 100, a separation mechanism installed inside the centrifuge 100, the separation mechanism including a first cylinder 200, a second cylinder 300 connected to the first cylinder 200, and a third cylinder 400 connected to the second cylinder 300. The primary sample is placed in the first cylinder 200. After one centrifugation, the sample in the first cylinder 200 enters the second cylinder 300. After a second centrifugation, the sample in the second cylinder 300 enters the third cylinder 400.

[0019] Collect the supernatant (sample) of cardiomyocytes in the logarithmic growth phase culture. Transfer the sample to the first cylinder 200, then install the second cylinder 300 on top of the first cylinder 200, and install the third cylinder 400 on top of the second cylinder 300. Install the three cylinders into the centrifuge 100 and start the centrifuge to make the three cylinders rotate synchronously, thereby centrifuging the sample to remove intact cardiomyocytes that have not adhered to the wall and large cell clusters. After the first centrifugation, stop the centrifuge 100 and transfer the supernatant in the first cylinder 200 to the second cylinder 300. Centrifuge again to remove broken cell debris, mitochondria and other large particulate impurities. After centrifugation, filter the supernatant in the second cylinder 300 into the third cylinder 400 to remove microvesicles and protein aggregates with a diameter greater than 220 nanometers. The sample pretreatment is now complete and can be further processed. The centrifuge 100 is equipped with a control panel 110, through which the centrifugation time and centrifugation speed are set; After the device is shut down, there is no need to change the centrifuge tubes multiple times. The sample can be transferred between the three tubes to achieve multiple centrifugation separations, thus improving work efficiency.

[0020] The first cylinder 200 has an internal thread at its port, and the second cylinder 300 has an external thread at its bottom that mates with the internal thread. The first cylinder 200, the second cylinder 300, and the third cylinder 400 are connected by a shaft 410 through their middle sections. The first piston plate 210 is slidably connected inside the first cylinder 200 and is connected to the bottom end of the shaft 410. The second piston plate 310 is slidably connected inside the second cylinder 300. An elastic pipe 220 connects the second piston plate 310 and the first piston plate 210 and is equipped with a one-way valve. The second piston plate 310 has a through hole in its middle that mates with the shaft 410. The first piston plate 210 is fixedly connected to the bottom end of the shaft 410. When the shaft 410 moves downward, the sample in the first cylinder 200 enters the second cylinder 300 through the elastic pipe 220.

[0021] After the first centrifugation is completed, the control shaft 410 moves downward. At this time, the shaft 410 drives the first piston plate 210 to move downward, so that the supernatant in the first cylinder 200 enters the second cylinder 300 through the elastic pipe 220 and is located in the upper space of the second piston plate 310. By controlling the downward movement distance of the first piston plate 210, the bottom sediment in the first cylinder 200 can be prevented from entering the second cylinder 300. The one-way valve prevents the sample in the second cylinder 300 from entering the first cylinder 200. After the sample is transferred to the second cylinder 300, a second centrifugation can be performed.

[0022] The top port of the second cylinder 300 is internally threaded to an annular frame 340 and externally threaded to the third cylinder 400. A filter membrane 350 is installed on the annular frame 340. When the second piston plate 310 moves upward, the sample after the second centrifugation can pass through the filter membrane 350 and enter the third cylinder 400.

[0023] After the second centrifugation, the second piston plate 310 is moved upward, pushing the centrifuged sample through the filter membrane 350 into the third cylinder 400. By controlling the upward movement distance of the second piston plate 310, large particles in the second cylinder 300 are prevented from entering the third cylinder 400.

[0024] A wedge 320 is radially slidably connected to the middle of the shaft 410. A spring 330 is fixedly connected between the wedge 320 and the shaft 410. After the shaft 410 moves down, the wedge 320 can pass through the second piston plate 310. After the wedge 320 passes through the second piston plate 310, the shaft 410 moves up, which can drive the second piston plate 310 to move up.

[0025] During the process of the shaft 410 moving downward to drive the upper clear liquid in the first cylinder 200 into the second cylinder 300, the wedge 320 can contact the second piston plate 310, and the wedge 320 is pushed by the inner wall of the second piston plate 310 to compress the spring 330 and slide into the shaft 410. At this time, the wedge 320 passes through the second piston plate 310, and then the spring 330 pushes out the wedge 320. Subsequently, when the shaft 410 moves upward, it can drive the second piston plate 310 to move upward. At this time, the sample after the second centrifugation enters the third cylinder 400.

[0026] Ventilation holes are provided on the side wall of the first cylinder 200 near the second cylinder 300, the side wall of the second cylinder 300 near the third cylinder 400, and the top wall of the third cylinder 400.

[0027] The vent holes ensure that the sliding of each piston plate is not affected by atmospheric pressure. Filter cotton or other filter media that can isolate external impurities and bacteria can be inserted into the vent holes to ensure that the sample inside the cylinder is not contaminated by the external environment.

[0028] A cover 120 is hinged to the centrifuge 100. The cover 120 has a circular hole coaxial with the third cylinder 400. Multiple electric telescopic rods 190 are radially slidably connected inside the circular hole. Each end of the electric telescopic rod 190 is fixedly connected to an L-shaped plate 191. Two ball bearings 192 are installed on the inner wall of the L-shaped plate 191. After the electric telescopic rod 190 is extended, the two ball bearings 192 roll on the top wall and side wall of the third cylinder 400, respectively.

[0029] The three electric telescopic rods 190 extend so that the L-shaped plate 191 can abut against the third cylinder 400. Since the ball bearing 192 is in contact with the outer wall of the third cylinder 400, the rotation of the third cylinder 400 is not interfered with during subsequent centrifugation, and the cylinder is firmly fixed in the centrifugation state.

[0030] A locking block 130 is rotatably connected to the cover 120, and a locking groove 140 is provided on the centrifuge 100 to cooperate with the locking block 130. After the locking block 130 is rotated, it can be locked into the locking groove 140.

[0031] The engagement of the locking groove 140 and the locking block 130 allows the cover 120 to be secured.

[0032] A pressure-sensitive switch is installed in the lock groove 140. When the lock block 130 is engaged in the lock groove 140, the pressure-sensitive switch can be pressed, thereby extending the multiple electric telescopic rods 190.

[0033] Once the locking block 130 is inserted into the locking groove 140, it indicates that the cover 120 is securely fixed. At this time, the locking block 130 applies pressure to the pressure-sensitive switch, thereby causing the electric telescopic rod 190 to extend. The device is then in place and ready for centrifugal extraction.

[0034] A cylinder 160 is fixedly connected to the cover 120. A mounting bracket 170 is fixedly connected to the output end of the cylinder 160. A locking block 180 is installed on the mounting bracket 170. A limit ring 420 is rotatably connected to the upper part of the shaft 410, and a cone block 430 is rotatably connected to the top. When the cover 120 is closed, the locking block 180 can be locked between the cone block 430 and the limit ring 420, so that the extension and retraction of the cylinder 160 can drive the shaft 410 to move axially.

[0035] The N-type frame 150 is fixedly connected to the cover 120, and the cylinder 160 is fixedly connected to the N-type frame 150. When the cover 120 is closed, the locking block 180 at the output end of the cylinder 160 can be locked between the limiting ring 420 and the cone block 430, so that when the cylinder 160 extends or retracts, it can drive the shaft 410 to move vertically.

[0036] The locking block 180 is elastically slidably connected to the mounting bracket 170.

[0037] The locking block 180 can also be elastically slidably connected to the mounting bracket 170 via a return spring. After centrifugation is completed, the locking block 180 can be manually pulled to disengage it from the limiting ring 420 and the cone block 430, thereby allowing the machine cover 120 to be opened.

[0038] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. An experimental apparatus for extracting exosomes, comprising a centrifuge (100), characterized in that: The centrifuge (100) is equipped with a separation mechanism, which includes a first cylinder (200), a second cylinder (300) connected to the first cylinder (200), and a third cylinder (400) connected to the second cylinder (300). The primary sample is placed in the first cylinder (200). After one centrifugation, the sample in the first cylinder (200) enters the second cylinder (300). After a second centrifugation, the sample in the second cylinder (300) enters the third cylinder (400).

2. The experimental apparatus for extracting exosomes according to claim 1, characterized in that: The first cylinder (200) has an internal thread at its port, and the second cylinder (300) has an external thread at its bottom that mates with the internal thread. The first cylinder (200), the second cylinder (300), and the third cylinder (400) are connected by a shaft (410) through their middle parts. The first cylinder (200) has a first piston plate (210) slidably connected inside, and the first piston plate (210) is connected to the bottom end of the shaft (410). The second cylinder (300) has a second piston plate (310) slidably connected inside, and an elastic pipe (220) connects the second piston plate (310) and the first piston plate (210). A one-way valve is provided on the elastic pipe (220). The second piston plate (310) has a through hole in the middle that cooperates with the shaft (410). The first piston plate (210) is fixedly connected to the bottom end of the shaft (410). When the shaft (410) moves down, the sample in the first cylinder (200) enters the second cylinder (300) through the elastic pipe (220).

3. The experimental apparatus for extracting exosomes according to claim 2, characterized in that: The top port of the second cylinder (300) is internally threaded to an annular frame (340) and externally threaded to the third cylinder (400). A filter membrane (350) is installed on the annular frame (340). When the second piston plate (310) moves upward, the sample after the second centrifugation can pass through the filter membrane (350) and enter the third cylinder (400).

4. The experimental apparatus for extracting exosomes according to claim 3, characterized in that: A wedge (320) is radially slidably connected to the middle of the shaft (410). A spring (330) is fixedly connected between the wedge (320) and the shaft (410). After the shaft (410) moves down, the wedge (320) can pass through the second piston plate (310). After the wedge (320) passes through the second piston plate (310), the shaft (410) moves up, which can drive the second piston plate (310) to move up.

5. The experimental apparatus for extracting exosomes according to claim 1, characterized in that: Ventilation holes are provided on the side wall of the first cylinder (200) near the second cylinder (300), the side wall of the second cylinder (300) near the third cylinder (400), and the top wall of the third cylinder (400).

6. The experimental apparatus for extracting exosomes according to claim 2, characterized in that: The centrifuge (100) is hinged to a cover (120), and the cover (120) has a circular hole coaxial with the third cylinder (400). Multiple electric telescopic rods (190) are radially slidably connected inside the circular hole. The ends of the multiple electric telescopic rods (190) are fixedly connected to L-shaped plates (191). Two ball bearings (192) are installed on the inner wall of the L-shaped plate (191). After the electric telescopic rods (190) are extended, the two ball bearings (192) roll on the top wall and side wall of the third cylinder (400) respectively.

7. The experimental apparatus for extracting exosomes according to claim 6, characterized in that: A locking block (130) is rotatably connected to the cover (120), and a locking groove (140) is provided on the centrifuge (100) to cooperate with the locking block (130). The locking block (130) can be engaged in the locking groove (140) after rotation.

8. The experimental apparatus for extracting exosomes according to claim 7, characterized in that: A pressure-sensitive switch is provided in the lock groove (140). When the lock block (130) is engaged in the lock groove (140), it can press the pressure-sensitive switch, thereby extending all of the electric telescopic rods (190).

9. The experimental apparatus for extracting exosomes according to claim 6, characterized in that: A cylinder (160) is fixedly connected to the cover (120), and a mounting bracket (170) is fixedly connected to the output end of the cylinder (160). A locking block (180) is installed on the mounting bracket (170). The upper part of the shaft (410) is rotatably connected to a limiting ring (420), and the top is rotatably connected to a cone block (430). When the cover (120) is closed, the locking block (180) can be locked between the cone block (430) and the limiting ring (420), so that the extension and retraction of the cylinder (160) can drive the shaft (410) to move axially.

10. The experimental apparatus for extracting exosomes according to claim 9, characterized in that: The card block (180) is elastically slidably connected to the mounting bracket (170).

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