Plasma exosome separation and extraction device

Abstract

The invention relates to the technical field of plasma exosome treatment, in particular to a plasma exosome separation and extraction device. The invention aims to solve the technical problems that when an ultracentrifugation method is used for extracting the plasma exosome, the operation is tedious, and the difficulty of operators is increased. According to the technical scheme, the plasma exosome separation and extraction device comprises a centrifugal machine body, the surface of the centrifugal machine body is rotationally connected with a sealing cover through a rotating shaft, and a controller is arranged on the surface of the centrifugal machine body and used for controlling operation in the centrifugal machine. The centrifugal machine has the beneficial effects that the reagent adding mechanism, the sample storage mechanism and the injection mechanism are matched for use, most of tedious operations are replaced by electric programming, an operator is prevented from repeatedly taking out a plasma sample from the centrifugal machine body, the plasma exosome separation and extraction time is shortened, and the operation efficiency is improved. And the operation difficulty of operators is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of plasma exosome processing, and in particular to a plasma exosome separation and extraction device. Background Art

[0002] Plasma exosomes are a specific extracellular vesicle with a diameter usually between 30-150 nanometers. They are released into the extracellular matrix after the fusion of intracellular multivesicular bodies with the cell membrane. Exosomes can participate in biological studies such as the body's immune response, antigen presentation, cell migration, cell differentiation and tumor invasion. Currently, there are many methods for the extraction of exosomes, such as ultracentrifugation, polymer sedimentation, ultrafiltration, etc. Among them, ultracentrifugation is the gold standard for exosome separation and the method is mature. However, the operation is cumbersome when separating and extracting exosomes, which increases the difficulty for operators.

[0003] Combining the above problems, we will find that it is difficult to avoid the above problems at the same time when using the existing plasma exosome separation and extraction devices on the market, and even if they can be solved, they need to be solved with the cooperation of external tools, which makes it impossible to achieve the desired effect. Therefore, we propose a plasma exosome separation and extraction device. Summary of the invention

[0004] The object of the present invention is to provide a plasma exosome separation and extraction device to solve the problems raised in the above background technology.

[0005] To achieve the above-mentioned object, the present invention provides the following technical solutions: a plasma exosome separation and extraction device, comprising a centrifuge body, a sealing cover is rotatably connected to the surface of the centrifuge body through a rotating shaft, a controller is arranged on the surface of the centrifuge body, and the controller is used to control the operation in the centrifuge, a centrifugal disk is rotatably connected to the inner cavity of the centrifuge body, and the centrifugal disk is used to rotate the sample at a high speed, six centrifugal grooves are opened on the surface of the centrifugal disk, and a reagent adding mechanism is arranged on the top of the centrifugal disk, and the reagent adding mechanism is used to add different reagents; The inner cavity of the centrifugal tank is provided with a sample storage mechanism, and the sample storage mechanism is used to store samples that need to be centrifuged, and the sample storage mechanism is used in conjunction with a reagent adding mechanism; The top of the centrifugal disk is provided with a push injection mechanism, which is used to push the sample, and the push injection mechanism is used in conjunction with the reagent adding mechanism and the sample storage mechanism.

[0006] Preferably, the reagent adding mechanism comprises a liquid storage box, the inner cavity of the liquid storage box is divided into a waste liquid area and a reagent area, the reagent area is arranged at the top of the waste liquid area, the inner wall of the liquid storage box is fixedly connected with a partition, the waste liquid area and the reagent area are divided by the partition, the top of the partition is fixedly connected with a positioning cylinder, the surface of the positioning cylinder is fixedly connected with four baffles, the four baffles divide the reagent area into four equal intervals, the four equal intervals are PBS area, TRIZOL area, chloroform area and isopropanol area in clockwise order, the inner cavity of the positioning cylinder is fixedly connected with a motor, the output end of the motor is fixedly connected with a first electric telescopic rod, the telescopic end of the first electric telescopic rod is fixedly connected with an L-shaped pressing plate, the inner cavities of the PBS area, TRIZOL area, chloroform area and isopropanol area are all slidably connected with first pistons, the tops of the four first pistons are fixedly connected with pressure rods, the L-shaped pressing plate is used in conjunction with the four pressure rods, and the surface of the liquid storage box is fixedly connected with two first Through tubes, one end of the two first through tubes are connected to the PBS area, the surface of the liquid storage box is fixedly connected with two second through tubes, one end of the two second through tubes are connected to the TRIZOL area, the other end of each of the first through tube and the second through tube is jointly provided with a first screw cover, the surface of the liquid storage box is fixedly connected with two third through tubes, one end of the two third through tubes are connected to the inner cavity of the chloroform area, the other end of each of the third through tubes is provided with a second screw cover, the surface of the liquid storage box is fixedly connected with two fourth through tubes, one end of the two fourth through tubes are connected to the isopropanol area, the other end of each of the fourth through tubes is provided with a third screw cover, the tops of the first screw cover, the second screw cover and the third screw cover are fixedly connected with a convex cylinder, the surface of the convex cylinder is rotatably connected with a cover cylinder through a sealing bearing, the other ends of the first through tube and the second through tube are jointly connected to the surface of a cover cylinder, and the third through tube and the fourth through tube are respectively fixedly connected to the surfaces of different cover cylinders.

[0007] Preferably, the surface of the liquid storage box is fixedly connected with two fifth through tubes, one end of the two fifth through tubes is connected with the waste liquid area, and the other end of the two fifth through tubes is fixedly connected with a flexible joint.

[0008] Preferably, the number of the sample storage mechanisms is six, and every three of the sample storage mechanisms form a group, and each group of the sample storage mechanisms includes a first tube, a second tube and a third tube, and the first tube, the second tube and the third tube are correspondingly distributed in three adjacent centrifugal grooves in a clockwise manner in sequence, the inner walls of the first tube, the second tube and the third tube are all slidably connected with piston blocks, the bottom of the piston block is fixedly connected with a push rod, and the surfaces of the three push rods are respectively slidably connected with the inner walls of the first tube, the second tube and the third tube, a waste liquid pipe is fixedly connected to the surface of the first tube, and the waste liquid pipe is connected to the fifth through pipe through a flexible joint, a sixth through pipe is fixedly connected to the surface of the first tube, the first tube and the second tube are connected through the sixth through pipe, a seventh through pipe is fixedly connected to the surface of the second tube, and the second tube and the third tube are connected through the seventh through pipe, and the tops of the first tube, the second tube and the third tube are all fixedly connected with threaded tubes, and the first screw cover, the second screw cover and the third screw cover are respectively threadedly connected to the surface of the threaded tube.

[0009] Preferably, a clamping block is fixedly connected to the inner wall of the centrifugal groove, and a limiting block is fixedly connected to the bottom of the push rod. The distance between the bottom of the clamping block and the bottom of the inner cavity of the centrifugal groove is equal to the thickness of the limiting block, and the limiting block is used in conjunction with the clamping block.

[0010] Preferably, a support block is provided on the surface of the seventh through tube, and the support block is fixedly connected to the top of the centrifugal disk. The inner wall of the support block is slidably connected to the surface of the seventh through tube. A locking block is provided on the surface of the seventh through tube, and a trapezoidal groove is provided on the inner wall of the locking block. The inner cavity of the trapezoidal groove is slidably connected to a limiting flow block, and the limiting flow block is used in conjunction with the seventh through tube.

[0011] Preferably, a spring is fixedly connected to one side of the current limiting block, and one end of the spring is fixedly connected to one side of the support block.

[0012] Preferably, the injection mechanism includes a second electric telescopic rod, the number of the second electric telescopic rod is four, the second electric telescopic rod is fixedly connected to the top of the centrifugal disk, the telescopic end of the second electric telescopic rod is fixedly connected to a positioning ring, the first tube and the second tube are respectively sleeved in the inner cavity of the positioning ring, the top of the positioning ring is fixedly connected to two mounting blocks, the inner sides of the two mounting blocks are respectively rotatably connected to the first rotating bar and the second rotating bar through a rotating shaft, one end of the first rotating bar is rotatably connected to the first clamping block through the rotating shaft, one end of the second rotating bar is rotatably connected to the second clamping block through the rotating shaft, a groove is provided on the surface of the first clamping block, and the surface of the second clamping block is in contact with the groove.

[0013] Preferably, the surfaces of the first tube and the second tube are both fixedly connected with clamping columns, and the clamping columns are used in conjunction with the clamping grooves.

[0014] Preferably, two L-shaped clamping rods are fixedly connected to the top of the positioning ring, and one side of the first rotating bar and the second rotating bar are both provided with a clamping slot, and the L-shaped clamping rods are used in conjunction with the clamping slots.

[0015] Compared with the prior art, the present invention has the following beneficial effects: The present invention utilizes a reagent adding mechanism to realize zoned storage of four reagents, and automatically adds required reagents according to operating steps, replacing manual addition by operators, thereby eliminating related cumbersome operating steps for adding reagents and reducing the operating difficulty for operators.

[0016] The present invention utilizes a sample storage mechanism and adopts a piston block and a push rod push injection type to replace a traditional integrated test tube, thereby facilitating subsequent push injection operations on plasma samples.

[0017] The present invention utilizes a push injection mechanism, and drives the first tube and the second tube to descend by contracting the second electric telescopic rod, thereby realizing the descent of the first tube and the second tube on the surface of the piston block and the push rod. When the first tube and the second tube descend, the plasma samples in the first tube and the second tube will rise under the action of the piston block, thereby realizing the push injection of the plasma sample. Through the coordinated use of the reagent adding mechanism, the sample storage mechanism and the push injection mechanism, most of the cumbersome operations are replaced by electric programming, avoiding the operator from repeatedly taking out the plasma sample from the centrifuge body, shortening the time for plasma exosome separation and extraction, and reducing the operating difficulty of the operator. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is a schematic diagram of the overall structure of the present invention; Figure 2 A top view of the inner cavity of the centrifuge body of the present invention; Figure 3 It is a three-dimensional schematic diagram of the relevant structure of the top of the centrifugal disk of the present invention; Figure 4 It is a cross-sectional schematic diagram of the reagent adding mechanism of the present invention; Figure 5 It is a three-dimensional schematic diagram of the push injection mechanism of the present invention; Figure 6 It is a schematic diagram of a cross-sectional three-dimensional exploded view of the sample storage mechanism of the present invention; Figure 7 is a three-dimensional schematic diagram of a support block of the present invention; Figure 8 It is a cross-sectional schematic diagram of the trapezoidal groove and the current limiting block of the present invention.

[0019] In the figure: 1, centrifuge body; 11, sealing cover; 12, controller; 13, centrifugal disc; 14, centrifugal tank; 2, reagent adding mechanism; 201, liquid storage box; 202, waste liquid area; 203, reagent area; 204, partition; 205, positioning cylinder; 206, baffle; 207, PBS area; 208, TRIZOL area; 209, chloroform area; 210, isopropanol area; 211, motor; 212, first electric telescopic rod; 213, L-shaped pressure plate; 214, first piston; 215, pressure rod; 216, first through pipe; 217, second through pipe; 218, first screw cover; 219, third through pipe; 220, second screw cover; 221, fourth through pipe; 222, third screw cover; 223, convex cylinder; 224, cover cylinder; 225, fifth through tube; 226, flexible joint; 3, sample storage mechanism; 31, first tube; 32, second tube; 33, third tube; 34, piston block; 35, push rod; 36, waste liquid tube; 37, sixth through tube; 38, seventh through tube; 39, threaded cylinder; 310, clamping block; 311, limiting block; 312, supporting block; 313, locking block; 314, trapezoidal groove; 315, current limiting block; 316, spring; 4, injection mechanism; 41, second electric telescopic rod; 42, positioning ring; 43, mounting block; 44, first rotating strip; 45, second rotating strip; 46, first clamping block; 47, second clamping block; 48, clamping groove; 49, clamping column; 410, L-shaped clamping rod; 411, bayonet. DETAILED DESCRIPTION

[0020] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] Example 1: Please refer to Figure 1-8 The present invention provides a technical solution: a plasma exosome separation and extraction device. The present invention makes corresponding improvements to the technical problems mentioned in the background technology, including a centrifuge body 1, a sealing cover 11 is rotatably connected to the surface of the centrifuge body 1 through a rotating shaft, a controller 12 is arranged on the surface of the centrifuge body 1, and the controller 12 is used to control the operation in the centrifuge, and a centrifugal disk 13 is rotatably connected to the inner cavity of the centrifuge body 1, and the centrifugal disk 13 is used to rotate the sample at a high speed, and six centrifugal grooves 14 are opened on the surface of the centrifugal disk 13, and a reagent adding mechanism 2 is arranged on the top of the centrifugal disk 13, and the reagent adding mechanism 2 is used to add different reagents.

[0022] As a further limitation of the reagent adding mechanism 2 of the present invention, the reagent adding mechanism 2 includes a liquid storage box 201, the inner cavity of the liquid storage box 201 is divided into a waste liquid area 202 and a reagent area 203, the reagent area 203 is arranged on the top of the waste liquid area 202, the inner wall of the liquid storage box 201 is fixedly connected with a partition 204, the waste liquid area 202 and the reagent area 203 are divided by the partition 204, the top of the partition 204 is fixedly connected with a positioning cylinder 205, the surface of the positioning cylinder 205 is fixedly connected with four baffles 206, the four baffles 206 divide the reagent area 203 into four equal intervals, and the four equal intervals are PBS area 207, TRIZOL area 208, chloroform area 209, and the like in a clockwise direction. 209 and isopropanol area 210, the inner cavity of the positioning cylinder 205 is fixedly connected with a motor 211, the output end of the motor 211 is fixedly connected with a first electric telescopic rod 212, the telescopic end of the first electric telescopic rod 212 is fixedly connected with an L-shaped pressing plate 213, the inner cavities of the PBS area 207, the TRIZOL area 208, the chloroform area 209 and the isopropanol area 210 are all slidably connected with first pistons 214, the tops of the four first pistons 214 are all fixedly connected with pressing rods 215, the L-shaped pressing plate 213 is used in conjunction with the four pressing rods 215, the surface of the liquid storage box 201 is fixedly connected with two first through pipes 216, one end of the two first through pipes 216 is connected with the PBS area 207, The surface of the liquid storage box 201 is fixedly connected with two second through-tubes 217, one end of the two second through-tubes 217 is connected to the TRIZOL area 208, and the other end of each first through-tube 216 and the second through-tube 217 is jointly provided with a first screw cap 218. The surface of the liquid storage box 201 is fixedly connected with two third through-tubes 219, one end of the two third through-tubes 219 is connected to the inner cavity of the chloroform area 209, and the other end of each third through-tube 219 is provided with a second screw cap 220. The surface of the liquid storage box 201 is fixedly connected with two fourth through-tubes 221, one end of the two fourth through-tubes 221 is connected to the isopropyl alcohol area 210, and the other end of each fourth through-tube 221 is provided with There is a third screw cover 222, and the tops of the first screw cover 218, the second screw cover 220 and the third screw cover 222 are all fixedly connected with a convex cylinder 223, and the surface of the convex cylinder 223 is rotatably connected with a cover cylinder 224 through a sealing bearing. The other ends of the first through tube 216 and the second through tube 217 are commonly connected to the surface of a cover cylinder 224, and the third through tube 219 and the fourth through tube 221 are respectively fixedly connected to the surfaces of different cover cylinders 224; by setting up a reagent adding mechanism 2, zoned storage of four reagents is realized, and the required reagents are automatically added according to the operating steps, instead of manual addition by the operator, which eliminates the related cumbersome operating steps of adding reagents and reduces the operating difficulty of the operator.

[0023] Two fifth through tubes 225 are fixedly connected on the surface of the liquid storage box 201, one end of the two fifth through tubes 225 is connected to the waste liquid area 202, and the other end of the two fifth through tubes 225 is fixedly connected to a flexible joint 226; by setting the fifth through tube 225, after the first ultracentrifugation is completed, the fifth through tube 225 can transfer the supernatant to be skimmed off to the waste liquid area 202, thereby realizing the first separation of the plasma sample.

[0024] The specific implementation of this embodiment is as follows: the operator needs to extract plasma exosomes, firstly, draw 1 ml of plasma sample and add it into the sample storage mechanism 3, position the sample storage mechanism 3 containing the plasma sample in the inner cavity of the centrifugal tank 14, seal the sample storage mechanism 3 through the first screw cover 218, the second screw cover 220 and the third screw cover 222, and position the sample storage mechanism 3 through the injection mechanism 4 at the same time, the operator closes the sealing cover 11 on the top of the centrifuge body 1, seals the centrifuge body 1, sets the operation steps through the controller 12, and then starts the operation, firstly, add 1 ml of PBS to each 1 ml of plasma sample, and in the initial state, the L-shaped pressing plate 213 is in the positive position of the PBS area 207 The first electric telescopic rod 212 is started by the program set by the controller 12 to shrink the first electric telescopic rod 212. The shrinkage of the first electric telescopic rod 212 drives the L-shaped pressing plate 213 to press downward. During the downward pressing process of the L-shaped pressing plate 213, the pressing rod 215 is pressed to drive the pressing rod 215 to move downward. The downward movement of the pressing rod 215 presses the first piston 214, so that the first piston 214 moves downward in the PBS area 207 to press the PBS stored in the PBS area 207. Quantitative solenoid valves are provided at the outlet ends of the first through-tube 216, the second through-tube 217, the third through-tube 219 and the fourth through-tube 221. When the first electric telescopic rod 212 is started for the first time, the second through-tube 217, the third through-tube 219 and the fourth through-tube 221 are started. A quantitative solenoid valve is arranged on the surface of the through pipe 216. The quantitative solenoid valve is an automatic control element with high precision and stability, and can accurately control the flow rate of the reagent. The PBS is pressed into the first through pipe 216 from the PBS area 207, and enters the plasma sample of the sample storage mechanism 3 through the first through pipe 216. When the flow rate of the PBS reaches 1 ml as monitored by the quantitative solenoid valve, the quantitative solenoid valve is closed, and at the same time, the telescopic end of the first electric telescopic rod 212 is controlled to extend to drive the L-shaped pressing plate 213 to reset. After the reset, the power supply of the first electric telescopic rod 212 is turned off, thereby realizing the quantitative injection of the PBS. Subsequently, the power supply of the centrifuge body 1 is started, and the centrifugal operation of 12000G is performed for 15 minutes according to the program setting. After the centrifugal operation of 15 minutes is completed, the plasma The sample is stratified, and the plasma sample is pushed upward for a distance by the push mechanism 4, so that the supernatant layer produced after centrifugation of the plasma sample completely enters the fifth through-tube 225, and enters the waste liquid area 202 through the fifth through-tube 225. The surface of the fifth through-tube 225 is provided with a solenoid valve. After the first centrifugation is completed, the solenoid valve is opened, and after the supernatant in the plasma sample is skimmed off, the solenoid valve is closed to prevent the skimmed supernatant from flowing back. While closing the solenoid valve, the motor 211 in the positioning cylinder 205 is started, and the rotation stroke of the motor 211 is a quarter of a turn clockwise. The rotation of the output end of the motor 211 drives the first electric telescopic rod 212 to rotate 90 degrees clockwise. At this time, the L-shaped pressing plate 213 is located above the pressing rod 215 of the TRIZOL area 208.At this time, the power of the motor 211 is turned off, and the first electric telescopic rod 212 is started, so that the telescopic end of the first electric telescopic rod 212 is contracted, and the L-shaped pressing plate 213 is driven to press the pressing rod 215 and the first piston 214 during the contraction, so that the TRIAOL in the TRIZOL area 208 enters the second through pipe 217 through pressure. At this time, the quantitative solenoid valve at the port of the second through pipe 217 is in an open state. When the TRIZOL reagent flowing through reaches 500 microliters, the quantitative solenoid valve is closed, and at the same time, the telescopic end of the first electric telescopic rod 212 is controlled to extend to drive the L-shaped pressing plate 213 to reset. After the reset, the power of the first electric telescopic rod 212 is turned off. After the injection of 500 microliters of TRIZOL reagent is completed, the centrifuge body has a temperature. The control function can accurately adjust the temperature during the centrifugation process. At this time, the temperature is adjusted to 4 degrees Celsius, and the TRIZOL reagent is reacted for 30 minutes. Then, the plasma sample is pushed up and down reciprocatingly through the push mechanism 4. During the push, the highest point where the plasma sample rises is lower than the inlet of the sixth through tube 37, thereby achieving mixing of the reacted plasma sample. Then, the mixed plasma is actually pushed upward by the push unit to enter the sixth through tube 37, and enters the new sample storage mechanism 3 through the sixth through tube 37. The surface of the sixth through tube 37 is provided with an electromagnetic valve. The electromagnetic valve on the surface of the sixth through tube 37 is opened at this time point through the controller 12 program setting. When the transfer of the serum sample in the first sample storage mechanism 3 is completed, the motor 2 is started. 11, the motor 211 continues to rotate clockwise 90 degrees, driving the L-shaped pressing plate 213 to rotate to the top of the pressing rod 215 in the chloroform area 209, and starting the first electric telescopic rod 212 to drive the L-shaped pressing plate 213 to move downward to press the pressing rod 215 and the first piston 214, so that the chloroform reagent in the chloroform area 209 enters the second sample storage mechanism 3 from the third through pipe 219. At this time, the quantitative solenoid valve at the port of the third through pipe 219 is in an open state. When the chloroform reagent flowing through reaches 100 microliters, the quantitative solenoid valve is closed, and the push injection mechanism 4 is started to move the plasma sample up and down, so that the plasma sample in the second sample storage mechanism 3 is fully mixed with the 100 microliters of chloroform reagent. At this time, the temperature in the centrifuge body 1 is adjusted to room temperature, and then it is left to stand for 3 minutes. Then, the centrifugal temperature of the centrifuge body 1 is adjusted to 4 degrees Celsius, 12000G, and centrifuged for 15 minutes. After the centrifugation is completed, the plasma sample is moved upward by the push injection mechanism 4, so that the supernatant produced after the centrifugation enters the seventh through tube 38. A flow sensor is arranged near the outlet end of the seventh through tube 38. The flow sensor can monitor the flow of the supernatant flowing through. When the flow of the supernatant flowing through reaches 200 microliters, the flow sensor controls the push injection mechanism 4 to move a slight distance in the reverse direction to avoid continuing to push the supernatant. 200 microliters of supernatant are pushed into the third sample storage mechanism 3. At this time, the motor 211 is started, and the motor 211 continues to rotate clockwise 90 degrees, driving the L-shaped pressing plate 213 to rotate to the top of the pressing rod 215 of the isopropanol area 210.The first electric telescopic rod 212 is started to drive the L-shaped pressing plate 213 to move downward to press the pressing rod 215 and the first piston 214, so that the isopropyl alcohol reagent in the isopropyl alcohol area 210 enters the third sample storage mechanism 3 from the fourth through pipe 221, and the injection amount of the isopropyl alcohol reagent is controlled to 200 microliters through the quantitative solenoid valve, and then the quantitative solenoid valve is closed, and at the same time, the first electric telescopic rod 212 is controlled to extend to reset the L-shaped pressing plate 213, and the motor 211 continues to rotate clockwise 90 degrees, so that the L-shaped pressing plate 213 returns to the top of the PBS area 207, and then the plasma sample with the isopropyl alcohol reagent is centrifuged at 12000G at a temperature of 4 degrees Celsius for 10 minutes, and then the centrifuged sample is taken out for subsequent operations.

[0025] Example 2: Please refer to Figure 1-8 The present invention provides a technical solution: a plasma exosome separation and extraction device. The present invention makes corresponding improvements to the technical problems mentioned in the background technology. The inner cavity of the centrifuge tank 14 is provided with a sample storage mechanism 3, and the sample storage mechanism 3 is used to store samples that need to be centrifuged. The sample storage mechanism 3 is used in conjunction with the reagent adding mechanism 2.

[0026] As a further limitation of the sample storage mechanism 3 of the present invention, the number of sample storage mechanisms 3 is six, and every three sample storage mechanisms 3 form a group. Each group of sample storage mechanisms 3 includes a first tube 31, a second tube 32, and a third tube 33. The first tube 31, the second tube 32, and the third tube 33 are correspondingly distributed in three adjacent centrifugal slots 14 in a clockwise manner. The inner walls of the first tube 31, the second tube 32, and the third tube 33 are all slidably connected with a piston block 34, and the bottom of the piston block 34 is fixedly connected with a push rod 35. The surfaces of the three push rods 35 are respectively slidably connected to the inner walls of the first tube 31, the second tube 32, and the third tube 33. The surface of the first tube 31 is fixedly connected with a waste liquid pipe 36, and the waste liquid pipe 36 is connected through a soft The connector 226 is connected to the fifth through tube 225, the surface of the first tube 31 is fixedly connected with the sixth through tube 37, the first tube 31 and the second tube 32 are connected through the sixth through tube 37, the surface of the second tube 32 is fixedly connected with the seventh through tube 38, the second tube 32 and the third tube 33 are connected through the seventh through tube 38, the tops of the first tube 31, the second tube 32 and the third tube 33 are fixedly connected with the threaded tube 39, the first screw cover 218, the second screw cover 220 and the third screw cover 222 are respectively threadedly connected to the surface of the threaded tube 39; by setting the sample storage mechanism 3 and adopting the piston block 34 and the push rod 35 push type, the traditional integrated test tube is replaced, which facilitates the subsequent push operation of the plasma sample.

[0027] A clamping block 310 is fixedly connected to the inner wall of the centrifugal tank 14, and a limiting block 311 is fixedly connected to the bottom of the push rod 35. The distance between the bottom of the clamping block 310 and the bottom of the inner cavity of the centrifugal tank 14 is equal to the thickness of the limiting block 311, and the limiting block 311 is used in conjunction with the clamping block 310. By setting the clamping block 310 and the limiting block 311, when the sample storage mechanism 3 is positioned in the centrifugal tank 14, the limiting block 311 is vertically moved downward to press against the bottom of the inner cavity of the centrifugal tank 14, and then the limiting block 311 is rotated to make the limiting block 311 embedded between the clamping block 310 and the centrifugal tank 14, thereby realizing the limitation of the sample storage mechanism 3.

[0028] The surface of the seventh through-tube 38 is provided with a support block 312, which is fixedly connected to the top of the centrifugal disc 13. The inner wall of the support block 312 is slidably connected to the surface of the seventh through-tube 38. The surface of the seventh through-tube 38 is provided with a locking block 313, and the inner wall of the locking block 313 is provided with a trapezoidal groove 314. The inner cavity of the trapezoidal groove 314 is slidably connected to a flow limiting block 315, and the flow limiting block 315 is used in conjunction with the seventh through-tube 38. By providing the support block 312, the sliding support of the seventh through-tube 38 is realized. The setting of the locking block 313 provides an area for the opening of the trapezoidal groove 314. When the centrifugal disk 13 rotates centrifugally, the locking block 313 will generate centrifugal movement on the surface of the seventh through tube 38. With the action of centrifugal force, the flow limiting block 315 will slide in the inner cavity of the trapezoidal groove 314. With the oblique trajectory of the trapezoidal groove 314, the flow limiting block 315 gradually clamps the surface of the seventh through tube 38. The seventh through tube 38 is a rubber hose, thereby achieving the purpose of limiting the flow of the seventh through tube 38 during centrifugation.

[0029] A spring 316 is fixedly connected to one side of the current limiting block 315, and one end of the spring 316 is fixedly connected to one side of the support block 312; by setting the spring 316, when centrifuging, the movement of the current limiting block 315 stretches the spring 316, thereby achieving elastic limitation of the moving distance of the current limiting block 315. When the centrifugal motion stops, the reaction force of the spring 316 drives the current limiting block 315 to reset.

[0030] The specific implementation of this embodiment is as follows: when positioning the sample storage mechanism 3, the limiting block 311 is moved downward vertically to press the limiting block 311 against the bottom of the inner cavity of the centrifugal tank 14, and then the limiting block 311 is rotated to make the limiting block 311 embedded between the clamping block 310 and the centrifugal tank 14, so as to achieve that before the perfusion of the plasma sample, the push rod 35 and the piston block 34 have been embedded in the inner cavity of the first tube 31, the second tube 32 and the third tube 33, and the push rod 35 is pulled down to drive the piston block 34 to move to the first tube 31, the second tube 32 and the third tube 33. 32 and the bottom of the inner cavity of the third tube 33, then add 1 ml of plasma sample, and after completing the embedding of the limit block 311 into the block 310, tighten the first screw cap 218 to the surface of the threaded tube 39 at the top of the first tube 31, tighten the second screw cap 220 to the surface of the threaded tube 39 at the top of the second tube 32, and tighten the third screw cap 222 to the surface of the threaded tube 39 at the top of the third tube 33, so as to achieve the closure of the first tube 31, the second tube 32 and the third tube 33, thereby ensuring the stability of the plasma sample during storage.

[0031] Example 3: Please refer to Figure 1-8 The present invention provides a technical solution: a plasma exosome separation and extraction device. The present invention makes corresponding improvements to the technical problems mentioned in the background technology. A push mechanism 4 is provided on the top of the centrifugal disk 13. The push mechanism 4 is used to push the sample. The push mechanism 4 is used in conjunction with a reagent adding mechanism 2 and a sample storage mechanism 3.

[0032] As a further limitation of the push injection mechanism 4 of the present invention, the push injection mechanism 4 includes a second electric telescopic rod 41, the number of the second electric telescopic rod 41 is four, the second electric telescopic rod 41 is fixedly connected to the top of the centrifugal disc 13, the telescopic end of the second electric telescopic rod 41 is fixedly connected to a positioning ring 42, the first tube 31 and the second tube 32 are respectively sleeved in the inner cavity of the positioning ring 42, the top of the positioning ring 42 is fixedly connected to two mounting blocks 43, the inner sides of the two mounting blocks 43 are respectively connected to the first rotating bar 44 and the second rotating bar 45 through the rotating shaft, and one end of the first rotating bar 44 is connected to the first clamping block through the rotating shaft 46, one end of the second rotating bar 45 is rotatably connected to the second clamping block 47 through a rotating shaft, a groove 48 is provided on the surface of the first clamping block 46, and the surface of the second clamping block 47 is in contact with the groove 48; by setting the push injection mechanism 4, the first tube 31 and the second tube 32 are driven to descend by the contraction of the second electric telescopic rod 41, so that the first tube 31 and the second tube 32 are lowered on the surface of the piston block 34 and the push rod 35. When the first tube 31 and the second tube 32 descend, the plasma samples in the first tube 31 and the second tube 32 will rise under the action of the piston block 34, thereby realizing the push injection of the plasma sample.

[0033] The surfaces of the first tube 31 and the second tube 32 are fixedly connected with a clamping column 49, which is used in conjunction with the clamping groove 48; by providing the clamping column 49, the clamping groove 48 is sleeved on the surface of the clamping column 49 when in use, thereby realizing vertical limitation and support for the first clamping block 46, and increasing the stability of the first clamping block 46 when positioned.

[0034] Two L-shaped clamping rods 410 are fixedly connected to the top of the positioning ring 42, and a bayonet 411 is provided on one side of the first turning bar 44 and the second turning bar 45, and the L-shaped clamping rod 410 is used in conjunction with the bayonet 411; by setting the L-shaped clamping rod 410 and the bayonet 411, when the first turning bar 44 and the second turning bar 45 drive the first clamping block 46 and the second clamping block 47 to clamp the first tube 31 and the second tube 32, the L-shaped clamping rod 410 is embedded in the inner cavity of the bayonet 411 to prevent the first turning bar 44 and the second turning bar 45 from continuing to rotate, thereby realizing the limitation of the rotation angle of the first turning bar 44 and the second turning bar 45.

[0035] The specific implementation of this embodiment is as follows: the positioning ring 42 is located just above the centrifugal groove 14, and the inner diameter of the positioning ring 42 is equal to the diameter of the centrifugal groove 14. When positioning the sample storage mechanism 3, the sample storage mechanism 3 is passed through the positioning ring 42 from top to bottom into the inner cavity of the centrifugal groove 14, and then the first rotating bar 44 and the second rotating bar 45 on the inner side of the two mounting blocks 43 are rotated in relative directions. The first rotating bar 44 and the second rotating bar 45 rotate to drive the first clamping block 46 and the second clamping block 47 to move relative to each other and move toward the surface of the first tube 31 and the second tube 32 at the same time. When the first clamping block 46 moves, the groove 48 on its surface will be embedded in the surface of the clamping column 49. The waste liquid pipe 36 is arranged on the surface of the first tube 31 opposite to the clamping column 49. The first clamp block 46 is supported and limited by the clamping column 49 and the waste liquid pipe 36. The surface of the second tube 32 is provided with two clamping columns 49 arranged opposite to each other to support the first clamp block 46. When fully clamped, one end of the second clamp block 47 is set to a rubber material. When it meets the first clamp block 46, it is embedded in the inner cavity of the clamping groove 48. Then, the second clamp block 47 and the first clamp block 46 are tightened by bolts to achieve a stable connection between the first clamp block 46 and the second clamp block 47. One end of the second clamp block 310 is embedded in the inner cavity of the clamping groove 48. In the initial state, the second electric telescopic rod 41 is in an extended state. When the plasma sample needs to be pushed, it can be achieved by contracting the second electric telescopic rod 41. When the sample needs to be mixed, it can be achieved by reciprocating the second electric telescopic rod 41.

[0036] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0037] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A plasma exosome separation and extraction device, comprising a centrifuge body (1), a sealing cover (11) being rotatably connected to the surface of the centrifuge body (1) via a rotating shaft, a controller (12) being arranged on the surface of the centrifuge body (1), the controller (12) being used to control the operation in the centrifuge, a centrifugal disc (13) being rotatably connected to the inner cavity of the centrifuge body (1), the centrifugal disc (13) being used to rotate a sample at a high speed, and six centrifugal grooves (14) being arranged on the surface of the centrifugal disc (13), characterized in that: A reagent adding mechanism (2) is provided on the top of the centrifugal disk (13), and the reagent adding mechanism (2) is used to add different reagents; The inner cavity of the centrifugal tank (14) is provided with a sample storage mechanism (3), the sample storage mechanism (3) is used to store samples that need to be centrifuged, and the sample storage mechanism (3) is used in conjunction with the reagent adding mechanism (2); A push mechanism (4) is provided on the top of the centrifugal disk (13), and the push mechanism (4) is used to push the sample, and the push mechanism (4) is used in conjunction with the reagent adding mechanism (2) and the sample storage mechanism (3); The reagent adding mechanism (2) comprises a liquid storage box (201), the inner cavity of the liquid storage box (201) is divided into a waste liquid area (202) and a reagent area (203), the reagent area (203) is arranged on the top of the waste liquid area (202), the inner wall of the liquid storage box (201) is fixedly connected to a partition (204), the waste liquid area (202) and the reagent area (203) are divided by the partition (204), the top of the partition (204) is fixedly connected to a positioning cylinder (205), the surface of the positioning cylinder (205) is fixedly connected to four baffles (206), the four baffles (206) divide the reagent area (203) into four equal intervals, and the four equal intervals are PBS areas in a clockwise direction. (207), a TRIZOL zone (208), a chloroform zone (209) and an isopropanol zone (210); the inner cavity of the positioning cylinder (205) is fixedly connected to a motor (211); the output end of the motor (211) is fixedly connected to a first electric telescopic rod (212); the telescopic end of the first electric telescopic rod (212) is fixedly connected to an L-shaped pressing plate (213); the inner cavities of the PBS zone (207), the TRIZOL zone (208), the chloroform zone (209) and the isopropanol zone (210) are all slidably connected to first pistons (214); the tops of the four first pistons (214) are all fixedly connected to pressing rods (215); the L-shaped pressing plate (213) and the four pressing rods (215) are connected to each other in a sliding manner. ), the surface of the liquid storage box (201) is fixedly connected with two first through tubes (216), one end of each of the two first through tubes (216) is connected to the PBS area (207), the surface of the liquid storage box (201) is fixedly connected with two second through tubes (217), one end of each of the two second through tubes (217) is connected to the TRIZOL area (208), the other end of each of the first through tubes (216) and the second through tubes (217) is jointly provided with a first screw cap (218), the surface of the liquid storage box (201) is fixedly connected with two third through tubes (219), one end of each of the two third through tubes (219) is connected to the inner cavity of the chloroform area (209), and each of the third through tubes The other end of each of the first and second screw caps (219) is provided with a second screw cap (220); the surface of the liquid storage box (201) is fixedly connected with two fourth through tubes (221); one end of each of the two fourth through tubes (221) is connected with the isopropyl alcohol zone (210); the other end of each of the fourth through tubes (221) is provided with a third screw cap (222); the tops of the first screw cap (218), the second screw cap (220) and the third screw cap (222) are fixedly connected with a convex cylinder (223); the surface of the convex cylinder (223) is rotatably connected with a cover cylinder (224) via a sealing bearing; the other ends of the first through tube (216) and the second through tube (217) are commonly connected to the surface of a cover cylinder (224);The third through pipe (219) and the fourth through pipe (221) are respectively fixedly connected to the surfaces of different cover tubes (224).

2. A plasma exosome separation and extraction device according to claim 1, characterized in that: The surface of the liquid storage box (201) is fixedly connected to two fifth through tubes (225), one end of each of the two fifth through tubes (225) is connected to the waste liquid area (202), and the other ends of each of the two fifth through tubes (225) are fixedly connected to a flexible joint (226).

3. A plasma exosome separation and extraction device according to claim 1, characterized in that: The number of the sample storage mechanisms (3) is six, and every three of the sample storage mechanisms (3) form a group. Each group of the sample storage mechanisms (3) comprises a first tube (31), a second tube (32) and a third tube (33). The first tube (31), the second tube (32) and the third tube (33) are sequentially and correspondingly distributed in three adjacent centrifugal grooves (14) in a clockwise direction. The inner walls of the first tube (31), the second tube (32) and the third tube (33) are all slidably connected to piston blocks (34). The bottom of the piston block (34) is fixedly connected to a push rod (35). The surfaces of the three push rods (35) are slidably connected to the inner walls of the first tube (31), the second tube (32) and the third tube (33). The surface of the first tube (31) is connected to the inner walls of the second tube (32) and the third tube (33). A waste liquid pipe (36) is fixedly connected to the surface of the first tube (31), and the waste liquid pipe (36) is connected to a fifth through pipe (225) through a flexible joint (226). A sixth through pipe (37) is fixedly connected to the surface of the first tube (31). The first tube (31) and the second tube (32) are connected through the sixth through pipe (37). A seventh through pipe (38) is fixedly connected to the surface of the second tube (32). The second tube (32) and the third tube (33) are connected through the seventh through pipe (38). The tops of the first tube (31), the second tube (32) and the third tube (33) are all fixedly connected to a threaded tube (39). The first screw cap (218), the second screw cap (220) and the third screw cap (222) are respectively threadedly connected to the surface of the threaded tube (39).

4. A plasma exosome separation and extraction device according to claim 3, characterized in that: A clamping block (310) is fixedly connected to the inner wall of the centrifugal groove (14), and a limiting block (311) is fixedly connected to the bottom of the push rod (35). The distance between the bottom of the clamping block (310) and the bottom of the inner cavity of the centrifugal groove (14) is equal to the thickness of the limiting block (311), and the limiting block (311) and the clamping block (310) are used in conjunction with each other.

5. The plasma exosome separation and extraction device according to claim 3, characterized in that: A support block (312) is provided on the surface of the seventh through tube (38), the support block (312) is fixedly connected to the top of the centrifugal disk (13), the inner wall of the support block (312) is slidably connected to the surface of the seventh through tube (38), a locking block (313) is provided on the surface of the seventh through tube (38), a trapezoidal groove (314) is provided on the inner wall of the locking block (313), the inner cavity of the trapezoidal groove (314) is slidably connected to a flow limiting block (315), and the flow limiting block (315) is used in conjunction with the seventh through tube (38).

6. A plasma exosome separation and extraction device according to claim 5, characterized in that: A spring (316) is fixedly connected to one side of the current limiting block (315), and one end of the spring (316) is fixedly connected to one side of the support block (312).

7. A plasma exosome separation and extraction device according to claim 3, characterized in that: The injection mechanism (4) comprises a second electric telescopic rod (41), the number of the second electric telescopic rod (41) is four, the second electric telescopic rod (41) is fixedly connected to the top of the centrifugal disc (13), the telescopic end of the second electric telescopic rod (41) is fixedly connected to a positioning ring (42), the first tube (31) and the second tube (32) are respectively sleeved in the inner cavity of the positioning ring (42), the top of the positioning ring (42) is fixedly connected to two mounting blocks (43), the inner sides of the two mounting blocks (43) are respectively rotatably connected to a first rotating bar (44) and a second rotating bar (45) via a rotating shaft, one end of the first rotating bar (44) is rotatably connected to a first clamping block (46) via a rotating shaft, one end of the second rotating bar (45) is rotatably connected to a second clamping block (47) via a rotating shaft, a surface of the first clamping block (46) is provided with a clamping groove (48), and a surface of the second clamping block (47) is in contact with the clamping groove (48).

8. A plasma exosome separation and extraction device according to claim 7, characterized in that: A clamping column (49) is fixedly connected to the surfaces of the first tube (31) and the second tube (32), and the clamping column (49) is used in conjunction with the clamping groove (48).

9. A plasma exosome separation and extraction device according to claim 8, characterized in that: Two L-shaped clamping rods (410) are fixedly connected to the top of the positioning ring (42); one side of each of the first rotating bar (44) and the second rotating bar (45) is provided with a clamping opening (411); the L-shaped clamping rods (410) are used in conjunction with the clamping opening (411).

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