Exosome extraction device and method

Abstract

The invention belongs to the technical field of exosome extraction, and particularly relates to an exosome extraction device and an extraction method.The exosome extraction device comprises a centrifugal machine, a reagent installation frame comprises a base connected with a driving motor, a vertical rod is fixed to the middle of the base, and a top base is fixed to the upper end of the vertical rod; a plurality of groups of symmetrically distributed reagent mounting seats are arranged on the lower side of the vertical rod, an adjusting assembly is arranged between the reagent mounting seats and the vertical rod, the plurality of groups of reagent mounting seats are uniformly distributed on the outer side of the vertical rod, and a sealing cover is arranged on the top seat; the multiple sealing covers are connected with the top base through connecting rods, channels are formed in the connecting rods, liquid inlets are formed in the bottoms of the sealing covers, a cavity is formed in the middle of the top base, and the multiple channels communicate with the cavity. By turning over the reagent mounting frame, the plurality of sample tubes are communicated with one another by utilizing a communicating vessel principle, so that the internal liquid reagents are balanced, and a worker does not need to balance the internal reagents of the plurality of sample tubes one by one, so that the working intensity is greatly reduced.

Description

Technical Field

[0001] The present invention belongs to the technical field of exosome extraction, and specifically relates to an exosome extraction device and an extraction method. Background Art

[0002] With the rapid development of biomedical technologies, exosomes, as important intercellular communication vectors, have shown tremendous potential in disease diagnosis, drug delivery, and regenerative medicine. Exosomes are rich in bioactive molecules such as proteins, nucleic acids (such as miRNA and mRNA), and lipids, and can reflect the physiological or pathological state of their cells of origin. Therefore, they have become a hot topic in the discovery of disease biomarkers and novel therapeutic strategies. However, the effective extraction and purification of exosomes is a key technical bottleneck that hinders their in-depth research and application.

[0003] Currently, methods for extracting exosomes primarily include ultracentrifugation, density gradient centrifugation, immunoaffinity capture, and polymer precipitation. Ultracentrifugation is widely used in both research and clinical practice due to its ease of use, relatively low cost, and ability to yield high-purity exosomes. Ultracentrifugation utilizes centrifugal force to separate the different components of a mixed solution based on density and particle size, enabling efficient extraction of exosomes.

[0004] During the ultracentrifugation extraction of exosomes, balancing the sample tubes is crucial. If there are mass differences between the sample tubes, unbalanced forces will be generated during the centrifugation process, causing increased vibration and noise in the centrifuge, and may even damage the centrifuge or cause a safety accident. Traditionally, staff need to manually balance multiple sample tubes one by one. This process is not only cumbersome and time-consuming, but also difficult to ensure the accuracy of balancing, which increases experimental errors and operational risks. Summary of the Invention

[0005] The purpose of the present invention is to provide an exosome extraction device and an extraction method to solve the problems raised in the background technology.

[0006] In order to achieve the above technical objectives, the technical solutions adopted by the present invention are as follows: An exosome extraction device includes a centrifuge, a working chamber is provided on the left side of the centrifuge, a drive motor is installed at the bottom of the working chamber, a reagent mounting rack connected to the drive motor is provided in the working chamber, the reagent mounting rack is used to mount sample tubes, the reagent mounting rack includes a base connected to the drive motor, a vertical rod is fixed in the middle of the base, and a top seat is fixed at the upper end of the vertical rod; Several groups of symmetrically distributed reagent mounting seats are provided on the lower side of the vertical pole, an adjustment component is provided between the reagent mounting seats and the vertical pole, several groups of the reagent mounting seats are evenly distributed on the outside of the column, and the top seat is provided with covers corresponding to the several reagent mounting seats, and the covers match the sample tubes; The plurality of covers are connected to the top seat by connecting rods, a channel is provided in the connecting rods, a liquid inlet is provided at the bottom of the cover and is connected to the channel, a cavity is provided in the middle of the top seat, and the plurality of channels are connected to the cavity; The sample tube is installed on the reagent installation seat, and an opening is provided on the lower side of the sample tube, and the opening is provided with a plug.

[0007] The sealing cover is provided with a sealing ring.

[0008] A blocking block is slidably provided in the cavity, and the blocking block is provided with a communication groove communicating with the two groups of channels symmetrically arranged.

[0009] A receiving groove is provided on the upper end of the top seat, a connecting block matching the receiving groove is fixed on the upper end of the blocking block, a magnet ring is connected to the lower side of the connecting block, and an iron ring matching the magnet ring is provided on the receiving groove.

[0010] The adjustment component includes a threaded section processed at the bottom of the vertical pole, a sliding sleeve threadedly connected to the threaded section is provided at the bottom of the vertical pole, a rotating ring is fixed to the upper end of the sliding sleeve, and the vertical pole sliding sleeve is provided with a fixed ring rotatably connected to the upper end of the rotating ring, and several of the reagent mounting seats are connected to the fixed ring via a fixed rod.

[0011] A sliding groove is provided on the side wall of the vertical rod, and a sliding block which slides in the sliding groove is fixed on the inner side of the fixing ring.

[0012] The side walls of the reagent mounting seat are provided with a plurality of elastic clips.

[0013] An extraction method of an exosome extraction device comprises the following steps: Step 1: Pour the liquid sample to be centrifuged into the sample tube, ensuring that the sample volume meets the experimental requirements; Step 2: Remove the reagent mounting rack from the working chamber and place it on the table with the base as the reference, ensuring that it is horizontal and stable. Insert the sample tube into the reagent mounting bracket and use the elastic clip to initially fix the bottom of the sample tube to prevent it from tipping over. Step 3: By rotating the sleeve, the threaded connection between the threaded section and the sleeve is utilized to move the sleeve upward, thereby pushing the reagent mounting seat and the sample tube upward until the upper end of the sample tube abuts against the cover; Step 4: Turn the reagent mounting rack upside down so that the top seat serves as the reference seat and is placed on the table. With the sample tube opening facing downward, remove the plug on the bottom side of the sample tube to connect the sample tube to the outside world. Using the communicating vessel principle, the liquid inside the sample tube will flow into the channel through the liquid inlet. Since all channels are connected to the cavity, all sample tubes will be interconnected, and the liquid level will remain the same, achieving sample tube balancing. Step 5: Re-attach the plug to the opening of the sample tube to ensure a seal. Flip the reagent mounting bracket to the right side so that the opening of the sample tube faces upward. The liquid in the channel will flow back into the respective sample tubes. Step 6: Place the balanced reagent mounting rack into the working chamber and connect it to the drive motor. Start the centrifuge and drive the reagent mounting rack to rotate at high speed through the drive motor to perform centrifugal work. Step 7: After centrifugation is complete, turn off the centrifuge and remove the reagent mounting rack after it stops rotating.

[0014] The present invention flips the reagent mounting rack and utilizes the communicating vessel principle to interconnect multiple sample tubes, thereby achieving internal liquid reagent balancing. Workers do not need to balance the reagents in multiple sample tubes one by one, which greatly reduces work intensity.

[0015] A blocking block is slidingly arranged in the cavity of the present invention. The blocking block is replaceable by cooperating with a magnet ring and an iron ring, and the number of connecting grooves of the blocking block can be selected in multiple ways. When centrifuging different numbers of sample tubes, the blocking blocks corresponding to the connecting grooves and the number of sample tubes can be replaced to achieve flexible balancing of different numbers of sample tubes. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The present invention is further illustrated by means of the following non-limiting examples.

[0017] Figure 1 This is a schematic structural diagram of an embodiment of an exosome extraction device and extraction method of the present invention; Figure 2 The structure of the reagent mounting rack of the present invention is shown as follows Figure 1 ; Figure 3 The structure of the reagent mounting rack of the present invention is shown as follows Figure 2 ; Figure 4 Schematic diagram of the cross-sectional structure of the reagent mounting rack of the present invention; Figure 5 for Figure 4 A schematic diagram of the structure at point A in the middle; Figure 6 It is a schematic diagram of the overall structure of the blocking block of the present invention.

[0018] The main component symbols are described as follows: Sample tube 0, plug 01, centrifuge 1, working chamber 11, reagent mounting rack 2, base 21, vertical rod 22, top seat 23, cavity 231, reagent mounting seat 24, elastic clip 241, cover 25, liquid inlet 251, sealing ring 252, connecting rod 26, channel 27, blocking block 3, connecting groove 31, receiving groove 32, connecting block 33, magnet ring 34, iron ring 35, threaded section 4, sliding sleeve 41, rotating ring 42, fixed ring 43, fixed rod 44, slide groove 45, slider 46. DETAILED DESCRIPTION

[0019] In order to enable those skilled in the art to better understand the present invention, the technical solution of the present invention is further described below with reference to the accompanying drawings and embodiments.

[0020] like Figure 1-6 As shown, an exosome extraction device of the present invention includes a centrifuge 1, a working chamber 11 is provided on the left side of the centrifuge 1, a driving motor is installed at the bottom of the working chamber 11, a reagent mounting rack 2 connected to the driving motor is provided in the working chamber 11, and the reagent mounting rack 2 is used to mount a sample tube 0, characterized in that: the reagent mounting rack 2 includes a base 21 connected to the driving motor, a vertical rod 22 is fixed in the middle of the base 21, and a top seat 23 is fixed at the upper end of the vertical rod 22; Several groups of symmetrically distributed reagent mounting seats 24 are provided on the lower side of the upright post 22. An adjustment assembly is provided between the reagent mounting seats 24 and the upright post 22. Several groups of reagent mounting seats 24 are evenly distributed on the outside of the upright post 22. The top seat 23 is provided with a cover 25 corresponding to the several reagent mounting seats 24. The cover 25 matches the sample tube 0. The plurality of covers 25 and the top seat 23 are connected to each other by connecting rods 26. A channel 27 is provided in the connecting rods 26. A liquid inlet 251 is provided at the bottom of the cover 25 and communicates with the channel 27. A cavity 231 is provided in the middle of the top seat 23. The plurality of channels 27 are communicated with the cavity 231. The sample tube 0 is installed on the reagent mounting seat 24 , and an opening is provided on the lower side of the sample tube 0 , and a plug 01 is provided at the opening.

[0021] Centrifuge 1 is an ultracentrifuge, an instrument that uses centrifugal force to quickly separate, precipitate and purify mixed solutions. Its rotation speed is extremely high, usually reaching tens of thousands of revolutions per minute or even higher. The centrifugal force generated is tens of thousands of times that of gravity, and it can separate substances with extremely small particle sizes and very small density differences; The drive motor is the power source of the ultracentrifuge. It can generate a strong driving force to drive the rotor (reagent mounting frame 2) to rotate at high speed. The motor is usually specially designed to withstand the huge centrifugal force, friction and heat generated during high-speed operation. The reagent mounting rack 2 in the present application is a horizontal rotor connected to a drive motor, and the sample reagent tube always remains in a horizontal state during the centrifugation process, which is conducive to the uniform sedimentation of the sample; When in use, the liquid sample is injected into the sample tube 0, and the staff takes out the reagent mounting rack 2 from the working chamber 11, and places the reagent mounting rack 2 on the desktop with the base 21 as the reference seat, and installs the sample tube 0 on the reagent mounting seat 24, and then drives the reagent mounting seat 24 upward through the adjustment component, so that the sample tube 0 moves upward and abuts against the cover 25, and the cover 25 can completely block the upper end of the sample tube 0, and the staff turns the reagent mounting rack 2 upside down, and the top seat 23 is placed on the desktop as the reference seat at this time, and the sample tube 0 is inverted at this time, and then the staff removes the plug 01 of the sample tube 0, so that The sample tube 0 is connected to the outside world. Therefore, the principle of communicating vessels is used between multiple sample tubes 0 to ensure that the liquid reagents inside the sample tubes 0 are the same. That is, the liquid sample inside the sample tube 0 will flow into the channel 27 through the liquid inlet 251. Since the uniform cavities 231 of the multiple channels 27 are connected, that is, the multiple sample tubes 0 are interconnected, the liquid levels of the multiple sample tubes 0 remain the same, so that the mass of the liquid reagents inside the multiple sample tubes 0 will be equal. After the staff re-fixes the plug 01 to the opening of the sample tube 0, and then flips the reagent mounting rack 2 to the forward direction, the liquid sample in the channel 27 will flow to the respective sample tubes 0. Through the above preparations, by turning over the reagent mounting rack 2 so that the openings of the sample tubes 0 face in the right direction, the multiple sample tubes 0 are connected to each other. By utilizing the principle of communicating vessels, the sample reagents in the multiple sample tubes 0 are balanced, eliminating the need for workers to balance the reagents in the multiple sample tubes 0 one by one, thus reducing work intensity. Finally, the reagent mounting rack 2 is placed in the working chamber 11 and connected to the driving motor, which drives the reagent mounting rack 2 to rotate at high speed to perform centrifugal work; The opening of the sample tube 9 and the plug 01 can be connected by a threaded connection to prevent the plug 01 from falling off during centrifugation.

[0022] The cover 25 is provided with a sealing ring 252 ; the sealing ring 252 can increase the sealing between the cover 25 and the sample tube 0 .

[0023] A blocking block 3 is slidably provided in the cavity 231 , and the blocking block 3 is provided with a communication groove 31 communicating with the two sets of channels 27 symmetrically arranged.

[0024] A receiving groove 32 is formed at the upper end of the top seat 23 , a connecting block 33 matching the receiving groove 32 is fixed to the upper end of the blocking block 3 , a magnet ring 34 is connected to the lower side of the connecting block 33 , and an iron ring 35 matching the magnet ring 34 is provided at the receiving groove 32 .

[0025] The sealing head 3 can be installed in the cavity 231 through the cooperation of the magnetic ring 34 and the iron ring 35, so the sealing block 3 can be replaced. The number of connecting grooves 31 of the sealing block 3 is even and symmetrical to each other, that is, the sealing block 3 with two connecting grooves 31, the sealing block 3 with four connecting grooves 31 or the sealing block 3 with six connecting grooves all correspond to the channels 27. Therefore, when centrifuging different numbers of sample tubes 0, it is necessary to replace the sealing blocks 3 with corresponding connecting grooves 31 to the number of sample tubes 0. For example, when centrifuging two sample tubes 0, the two sample tubes 0 are symmetrically placed on the reagent mounting rack 0, and the sealing block 3 with two connecting grooves 31 is taken and slid into the cavity 231. The two connecting grooves 31 of the sealing block 31 will correspond to the two channels 27 and connect the two channels 27. The remaining channels 27 will be blocked by the side walls of the sealing block 31. In this way, only the two sample tubes 0 can be connected to each other for balancing. The provision of the receiving groove 32 enables the blocking block 3 as a whole to be flush with the upper end surface of the top seat 23 , ensuring that the top seat 23 can be placed flat on the desktop.

[0026] The adjustment component includes a threaded section 4 processed at the bottom of the vertical rod 22, a sliding sleeve 41 threadedly connected to the threaded section 4 is provided at the bottom of the vertical rod 22, a rotating ring 42 is fixed to the upper end of the sliding sleeve 41, and the sliding sleeve of the vertical rod 22 is provided with a fixed ring 43 rotatably connected to the upper end of the rotating ring 42, and several reagent mounting seats 24 are connected to the fixed ring 43 through a fixed rod 44; after the sample tube 0 is inserted into the reagent mounting seat 24, the sliding sleeve 41 is rotated. Since the sliding sleeve 41 is threadedly connected to the threaded section 4 at the bottom of the vertical rod 22, the sliding sleeve 41 will move upward, and the rotating sleeve 42 drives the fixed ring 42 to move upward, thereby driving the sample tube 0 to move upward and abut against the cover.

[0027] A sliding groove 45 is provided on the side wall of the vertical rod 22, and a slider 46 sliding in the sliding groove 45 is fixed to the inner side of the fixed ring 43; the fixed ring 43 slides in the sliding groove 45 through the slider 46, so that it can only move up and down along the vertical rod 22, and when the sliding sleeve 41 moves upward, it will drive the rotating ring 42 to rotate. Since the fixed ring 43 is rotatably connected to the rotating ring 42, the rotation of the rotating ring 42 will not drive the fixed ring 43 to rotate, and can only push it to move upward.

[0028] The sidewall of the reagent mounting seat 24 is provided with a plurality of elastic clips 241. The elastic clips 241 are used to initially clamp the bottom of the sample tube 0 so that it does not fall over.

[0029] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the present invention. Anyone skilled in the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by one of ordinary skill in the art without departing from the spirit and technical principles disclosed herein are intended to be covered by the claims of the present invention.

Claims

1. An exosome extraction device, comprising a centrifuge, a working chamber provided on the left side of the centrifuge, a drive motor installed at the bottom of the working chamber, a reagent mounting rack connected to the drive motor provided in the working chamber, the reagent mounting rack being used to mount sample tubes, characterized in that: The reagent mounting rack includes a base connected to the drive motor, a vertical rod is fixed in the middle of the base, and a top seat is fixed at the upper end of the vertical rod; Several groups of symmetrically distributed reagent mounting seats are provided on the lower side of the vertical pole, an adjustment component is provided between the reagent mounting seats and the vertical pole, several groups of the reagent mounting seats are evenly distributed on the outside of the column, and the top seat is provided with covers corresponding to the several reagent mounting seats, and the covers match the sample tubes; The plurality of covers are connected to the top seat by connecting rods, a channel is provided in the connecting rods, a liquid inlet is provided at the bottom of the cover and is connected to the channel, a cavity is provided in the middle of the top seat, and the plurality of channels are connected to the cavity; The sample tube is installed on the reagent installation seat, and an opening is provided on the lower side of the sample tube, and the opening is provided with a plug.

2. The exosome extraction device according to claim 1, characterized in that: The sealing cover is provided with a sealing ring.

3. The exosome extraction device according to claim 2, characterized in that: A blocking block is slidably provided in the cavity, and the blocking block is provided with a communication groove communicating with the two groups of channels symmetrically arranged.

4. The exosome extraction device according to claim 3, characterized in that: A receiving groove is provided on the upper end of the top seat, a connecting block matching the receiving groove is fixed on the upper end of the blocking block, a magnet ring is connected to the lower side of the connecting block, and an iron ring matching the magnet ring is provided on the receiving groove.

5. The exosome extraction device according to claim 4, characterized in that: The adjustment component includes a threaded section processed at the bottom of the vertical pole, a sliding sleeve threadedly connected to the threaded section is provided at the bottom of the vertical pole, a rotating ring is fixed to the upper end of the sliding sleeve, and the vertical pole sliding sleeve is provided with a fixed ring rotatably connected to the upper end of the rotating ring, and several of the reagent mounting seats are connected to the fixed ring via a fixed rod.

6. The exosome extraction device according to claim 5, characterized in that: A sliding groove is provided on the side wall of the vertical rod, and a sliding block which slides in the sliding groove is fixed on the inner side of the fixing ring.

7. The exosome extraction device according to claim 6, characterized in that: The side walls of the reagent mounting seat are provided with a plurality of elastic clips.

8. The method for extracting exosomes according to claim 1-7, characterized in that: The following steps are involved: Step 1: Pour the liquid sample to be centrifuged into the sample tube, ensuring that the sample volume meets the experimental requirements; Step 2: Remove the reagent mounting rack from the working chamber and place it on the table with the base as the reference, ensuring that it is horizontal and stable. Insert the sample tube into the reagent mounting bracket and use the elastic clip to initially fix the bottom of the sample tube to prevent it from tipping over. Step 3: By rotating the sleeve, the threaded connection between the threaded section and the sleeve is utilized to move the sleeve upward, thereby pushing the reagent mounting seat and the sample tube upward until the upper end of the sample tube abuts against the cover; Step 4: Turn the reagent mounting rack upside down so that the top seat serves as the reference seat and is placed on the table. With the sample tube opening facing downward, remove the plug on the bottom side of the sample tube to connect the sample tube to the outside world. Using the communicating vessel principle, the liquid inside the sample tube will flow into the channel through the liquid inlet. Since all channels are connected to the cavity, all sample tubes will be interconnected, and the liquid level will remain the same, achieving sample tube balancing. Step 5: Re-attach the plug to the opening of the sample tube to ensure a seal. Flip the reagent mounting bracket to the right side so that the opening of the sample tube faces upward. The liquid in the channel will flow back into the respective sample tubes. Step 6: Place the balanced reagent mounting rack into the working chamber and connect it to the drive motor. Start the centrifuge and drive the reagent mounting rack to rotate at high speed through the drive motor to perform centrifugal work. Step 7: After centrifugation is complete, turn off the centrifuge and remove the reagent mounting rack after it stops rotating.

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