Flow pretreatment device and method for detecting surface antigens and intracellular proteins

Abstract

This invention relates to the field of flow cytometry pre-sample processing, specifically to a flow cytometry pre-processing apparatus and method for detecting surface antigens and intracellular proteins. The apparatus includes a support plate, a base, four lifting and vibrating synchronous mechanisms, and a synchronous drive mechanism. Each lifting and vibrating synchronous mechanism includes a vertical plate, a corrugated plate, a first roller, a lifting drive structure, and a vibration transmission structure. The corrugated plate is located on one side of the upper part of the vertical plate, the lifting drive structure is located on the other side of the upper part of the vertical plate, and the vibration transmission structure is located between the first roller and the support plate. The synchronous drive mechanism includes a second rotating shaft, four transmission components, and a rotation drive structure. The four transmission components simultaneously drive the four lifting and vibrating synchronous mechanisms, and the rotation drive structure drives the second rotating shaft to rotate. This invention, by setting up four lifting and vibrating synchronous mechanisms and a synchronous drive mechanism, ensures that the solution is uniformly mixed and immediately placed within the magnetic separation mechanism, eliminating the time required to transfer the test tubes to the magnetic separation mechanism.

Description

Technical Field

[0001] The present invention relates to the field of pre-flow sample processing, and in particular to a pre-flow processing device for detecting surface antigens and intracellular proteins, and in particular to a method for using the pre-flow processing device for detecting surface antigens and intracellular proteins. Background Art

[0002] Flow pretreatment devices play an important role in many fields, such as food processing and biomedical research. These devices pre-treat samples or raw materials through specific processes and technologies for subsequent analysis, detection or processing. Flow pretreatment devices refer to equipment or systems that can automatically or manually complete sample pretreatment steps. In the biomedical field, flow pretreatment devices are often used in the sample preparation stage of flow cytometry.

[0003] The patent with announcement number CN114713367B discloses an oscillating magnetic separation device and method for a flow pre-sample processing instrument. The working principle is as follows: open the top cover and place the microporous tube containing the biological sample and the specifically bound magnetic particle solution into the microporous tray; energize the double-headed motor to drive the eccentric wheel to rotate, and under the action of the limit mechanism, the oscillating disk can reciprocate and make a semi-rotational motion, which can drive the first shell and the second shell to vibrate as a whole, so that the biological sample and the specifically bound magnetic particle solution in the microporous tube can be fully mixed; the first drive motor is energized to drive the worm to rotate, so that the turbine rotates to drive the synchronous shaft to rotate, and the connecting rod rotates along the top The lifting plate is flipped in the lifting direction, so that the lifting rod can drive the lifting plate to rise along the microporous tray, and the electromagnetic rod is also inserted into the gap of the microporous tube; the second driving motor is energized to drive the second bevel gear to rotate, and under the action of the kinetic energy transmission of the second bevel gear, the first bevel gear drives the second gear to rotate through the transmission shaft. At the same time, the first gear drives the bracket to rotate, so that the electromagnetic rod can rotate around the microporous tube. Under continuous rotation, the magnetic particles form a wavy motion trajectory in the microporous tube, so that the magnetic particles in the solution carry the bound target substances and are adsorbed to the side wall to achieve side wall agglomeration. After completing the magnetic separation of the target substance and the non-target substance, the bracket is lowered by the lifting mechanism.

[0004] The following problems still exist in the above patent document: the second shell and the first shell of the oscillating plate drive the microporous tray to vibrate, the microporous tray drives the microporous tube to vibrate, the biological samples in the microporous tube are mixed evenly, and then several electromagnetic rods are raised upward to make the microporous tube within the rotation coverage range of several electromagnetic rods. The two working steps are completely separated, which makes the sample processing time longer, resulting in reduced efficiency of flow detection. Summary of the invention

[0005] In view of the above problems, a flow pretreatment device for detecting surface antigens and intracellular proteins is provided. The present invention is provided with four lifting and vibration synchronization mechanisms and a synchronous driving mechanism, so that the solution is placed in the magnetic separation mechanism after being evenly mixed, thereby eliminating the time of transferring the test tube to the magnetic separation mechanism.

[0006] In order to solve the problems of the prior art, the present invention provides a flow pretreatment device for detecting surface antigens and intracellular proteins, comprising a carrying plate for placing a test tube holder and a base for installing a magnetic separation mechanism, and also comprising four lifting and vibration synchronization mechanisms and a synchronous drive mechanism; the four lifting and vibration synchronization mechanisms are respectively arranged in the middle of the four sides of the carrying plate, the lifting and vibration synchronization mechanism comprises a vertical plate, a wave plate, a first roller, a lifting drive structure and a vibration transmission structure, the lower end of the vertical plate is connected to the base, the wave plate is arranged on one side of the upper part of the vertical plate, the first roller rolls along the wave plate, a first rotating shaft is arranged in the first roller, and the first roller is extended from both ends of the first rotating shaft, the lifting drive structure is arranged on the other side of the upper part of the vertical plate, and the vibration transmission structure is arranged between the first roller and the carrying plate; the synchronous drive mechanism is arranged at the lower end of the base, the synchronous drive mechanism comprises a second rotating shaft, four transmission components and a rotation drive structure, the second rotating shaft is vertically arranged in the middle of the base, the four transmission components are annularly arrayed around the second rotating shaft, the four transmission components are used to simultaneously drive the four lifting and vibration synchronization mechanisms to operate, and the rotation drive structure is used to drive the second rotating shaft to rotate.

[0007] Preferably, the vibration transmission structure includes a connecting plate, a first connecting column, a compression block and a first spring; the connecting plate is connected to the load-bearing plate; there are a plurality of first connecting columns, which are evenly distributed at both ends of the connecting plate, and one end of the first connecting column is connected to the connecting plate; there are two compression blocks, which are respectively arranged at both ends of the first rotating shaft, and both ends of the compression blocks are plugged into the first connecting column; there are a plurality of first springs, which are respectively mounted on a plurality of first connecting columns, and both ends of the first spring are respectively abutted against the connecting plate and the compression block.

[0008] Preferably, the lifting drive structure includes a screw, a lifting block and two connecting claws; the screw is vertically arranged on the vertical plate; the middle part of the lifting block is threadedly connected to the screw, and the screw rotates to drive the lifting block to move along the axial direction of the screw; the two connecting claws are respectively arranged at both ends of the lifting block, one end of the connecting claw is connected to the lifting block, and the other end of the connecting claw is provided with a strip groove, and the end of the first rotating shaft is slidably arranged in the strip groove.

[0009] Preferably, the lifting and vibration synchronization mechanism also includes two stabilizing structures, which are respectively arranged on both sides of the vertical plate, and the stabilizing structures include a first guide structure and two balancing connecting arms; the first guide structure includes a first guide rod and a first slider, the first guide rod is arranged parallel to the screw rod, and the first slider is slidably arranged on the first guide rod; the two balancing connecting arms are arranged in parallel, one end of the balancing connecting arm is connected to the end of the first slider, and the two balancing connecting arms are abutted against the upper and lower surfaces of the connecting claws, and the other end of the balancing connecting arm is connected to the supporting plate.

[0010] Preferably, the stabilizing structure further includes a vibration buffer structure, which is disposed on the bearing plate and is used to connect the balancing connecting arm and the bearing plate.

[0011] Preferably, the lifting and vibration synchronization mechanism also includes an ejection structure, which is arranged in the middle of the lifting block, and the ejection structure includes a first bracket, a push rod, a curved top plate and a second spring; the first bracket is connected to the lifting block; the push rod is slidably connected to the first bracket, and the push rod can move along its axial direction; the curved top plate is arranged at one end of the push rod, and the inner concave surface of the curved top plate abuts against the middle of the first roller; the second spring is sleeved on the other end of the push rod, and the two ends of the second spring respectively abut against the end of the push rod and the first bracket.

[0012] Preferably, a plurality of second rollers are arranged on the inner concave side of the arc-shaped top plate, and the plurality of second rollers are all tangent to the first roller.

[0013] Preferably, the rotary drive structure includes a gear, a rectangular frame, a second guide structure, a continuous transmission structure, a third rotating shaft, a transmission connecting rod assembly and a U-shaped frame; the gear is connected to the second rotating shaft and a rectangular frame cover is arranged on the outside of the gear; the second guide structure has at least two, and the two second guide structures are respectively arranged on both sides of the rectangular frame, and the second guide structure is used to limit the moving direction of the rectangular frame; there are two continuous transmission structures, and the two continuous transmission structures are symmetrically arranged on both sides of the gear; the third rotating shaft is vertically arranged at one end of the rectangular frame; the transmission connecting rod assembly is arranged between the third rotating shaft and the rectangular frame, and the transmission connecting rod assembly is used to convert the rotation of the third rotating shaft into reciprocating linear motion of the rectangular frame along the guide direction of the second guide structure; both ends of the U-shaped frame are connected to the base, and the middle part of the third rotating shaft is connected to the U-shaped frame through a bearing.

[0014] Preferably, the rotary drive structure further includes a linkage structure, and the linkage structure is arranged between the magnetic separation mechanism and the third rotating shaft.

[0015] The method for using a flow pre-treatment device for detecting surface antigens and intracellular proteins comprises the following steps:

[0016] S1, firstly put a plurality of test tubes containing biological samples and specifically bound magnetic particle solutions into a test tube holder, and then fix the test tube holder on a carrying plate;

[0017] S2, the rotary drive structure drives the second rotating shaft to rotate, and the second rotating shaft drives the four lifting drive structures to operate simultaneously through four transmission components;

[0018] S3, the lifting drive structure drives the first roller to move from top to bottom along the corrugated plate;

[0019] S4, when the first roller rolls along the wave plate, the distance between the first roller and the bearing plate changes repeatedly, the vibration transmission structure is repeatedly compressed and released, and the vibration generated by the compression and release is transmitted to the bearing plate, and the bearing plate drives the test tube to vibrate;

[0020] S5, after the first roller rolls to the lower end of the wave plate, the biological sample in the test tube and the specifically bound magnetic particle solution are fully mixed, and all the test tubes are located in the magnetic separation mechanism. The continuous rotation of the magnetic separation mechanism causes the magnetic particles in the solution to carry the bound target substances to be adsorbed to the side wall to achieve side wall aggregation, thereby completing the magnetic separation of the target substance and the non-target substance.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] 1. The present invention is provided with four lifting and vibration synchronization mechanisms and a synchronization driving mechanism. The synchronization driving mechanism drives the four lifting and vibration synchronization mechanisms to operate simultaneously, so that the carrier plate can be lifted and lowered smoothly. The lifting driving structure drives the first roller to roll from top to bottom along the wave plate, so that the carrier plate maintains horizontal vibration to avoid spilling of the solution in the test tube. After the first roller rolls to the lower end of the wave plate, the biological sample in the test tube and the specifically bound magnetic particle solution are fully mixed, and all the test tubes are located within the working range of the magnetic separation mechanism. The continuous rotation of the magnetic separation mechanism allows the magnetic particles in the solution to carry the bound target substances and be adsorbed to the side wall to achieve side wall agglomeration, thereby completing the magnetic separation of the target substance and the non-target substance, thereby achieving that the solution is evenly mixed and is in the magnetic separation mechanism, and the time for transferring the test tube to the magnetic separation mechanism is omitted.

[0023] 2. The first roller of the present invention pushes the compression block to move along the first connecting column toward the connecting plate through the first rotating shaft. The first spring between the compression block and the connecting plate is compressed, and the force exerted by the first roller on the compression block toward the connecting plate is reduced. The first spring releases elastic potential energy to push the compression block along the first connecting column away from the first connecting plate. The first spring is repeatedly compressed and released at a fixed frequency, thereby generating vibrations, which are transmitted to the load-bearing plate through the connecting plate, thereby realizing repeated vibrations of the first spring at a fixed frequency, avoiding vibration superposition that causes an increase in vibration amplitude, and maintaining the vibration frequency of the test tube.

[0024] 3. The present invention is provided with two stabilizing structures, one end of the balancing connecting arm is connected to the first slider, and the other end is connected to the supporting plate. Meanwhile, the two balancing connecting arms are in close contact with the upper and lower surfaces of the connecting claws. With the rotation of the screw rod, the lifting block starts to move up and down through the connecting claws. Since the connecting claws are connected to the balancing connecting arms, the supporting plate is not only subjected to direct force from the lifting block and the vibration transmission structure during the lifting process, but also obtains additional stability support through the balancing connecting arms, thereby effectively reducing the lag and unevenness in the force transmission process, and making the four sides of the supporting plate more evenly stressed. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is a stereoscopic diagram of a flow cytometry pretreatment device for detecting surface antigens and intracellular proteins.

[0026] Figure 2 This is a front view of a flow cytometry pretreatment device for detecting surface antigens and intracellular proteins.

[0027] Figure 3 yes Figure 2 Stereoscopic cross-sectional view at AA in the middle.

[0028] Figure 4 It is a stereoscopic diagram of a lifting and vibrating synchronization mechanism in a flow pre-treatment device for detecting surface antigens and intracellular proteins.

[0029] Figure 5 It is a stereoscopic diagram of a wave plate, a first roller and a vibration transmission structure in a flow pretreatment device for detecting surface antigens and intracellular proteins.

[0030] Figure 6 It is a stereoscopic diagram of a wave plate, a first roller and a lifting drive structure in a flow pretreatment device for detecting surface antigens and intracellular proteins.

[0031] Figure 7 It is a three-dimensional diagram of the lifting block, connecting claws and stabilizing structure in the flow pretreatment device for detecting surface antigens and intracellular proteins.

[0032] Figure 8 It is a three-dimensional diagram of a carrier plate and a stabilizing structure in a flow pretreatment device for detecting surface antigens and intracellular proteins.

[0033] Fig. 9 It is a stereoscopic diagram of a first roller, a lifting block and an ejection structure in a flow pretreatment device for detecting surface antigens and intracellular proteins.

[0034] Fig.10 It is a stereoscopic diagram of a first roller, a curved top plate and a second roller in a flow pretreatment device for detecting surface antigens and intracellular proteins.

[0035] Fig.11It is a stereoscopic diagram of a magnetic separation mechanism, a base and a synchronous drive mechanism in a flow pretreatment device for detecting surface antigens and intracellular proteins.

[0036] Fig.12 It is a stereoscopic diagram of a second rotating shaft and a rotating drive structure in a flow pretreatment device for detecting surface antigens and intracellular proteins.

[0037] Fig.13 It is a stereoscopic diagram of a magnetic separation mechanism, a third rotating shaft and a linkage structure in a flow pretreatment device for detecting surface antigens and intracellular proteins.

[0038] The numbers in the figure are: 1. test tube holder; 2. bearing plate; 3. magnetic separation mechanism; 4. base; 5. lifting and vibration synchronization mechanism; 51. vertical plate; 52. wave plate; 53. first roller; 531. first rotating shaft; 54. lifting drive structure; 541. screw rod; 542. lifting block; 543. connecting claw; 5431. strip groove; 55. vibration transmission structure; 551. connecting plate; 552. first connecting column; 553. compression block; 554. first spring; 56. stabilizing structure; 561. first guide structure; 5611. first guide rod; 5612. first slider; 562. balancing connecting arm; 563. vibration buffer structure; 5631. second guide rod; 5632. second slider; 57. ejector structure; 571, first bracket; 572, top rod; 573, arc top plate; 5731, second roller; 574, second spring; 575, linear drive unit; 6, synchronous drive mechanism; 61, second rotating shaft; 62, transmission assembly; 621, first transmission shaft; 622, second transmission shaft; 63, rotation drive structure; 631, gear; 632, rectangular frame; 633, second guide structure; 6331, third guide rod; 6332, third slider; 634, continuous transmission structure; 6341, transmission rack; 6342, linear drive; 635, third rotating shaft; 636, transmission connecting rod assembly; 637, linkage structure; 6371, transmission wheel; 6372, transmission belt; 638, U-shaped frame. DETAILED DESCRIPTION

[0039] In order to further understand the features, technical means, specific objectives and functions of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation methods.

[0040] Reference Figures 1 to 13As shown: a flow pretreatment device for detecting surface antigens and intracellular proteins, comprising a carrier plate 2 for placing a test tube holder 1 and a base 4 for installing a magnetic separation mechanism 3, and also comprising four lifting and vibration synchronization mechanisms 5 and a synchronization drive mechanism 6; the four lifting and vibration synchronization mechanisms 5 are respectively arranged at the middle of the four sides of the carrier plate 2, and the lifting and vibration synchronization mechanism 5 comprises a vertical plate 51, a wave plate 52, a first roller 53, a lifting drive structure 54 and a vibration transmission structure 55, the lower end of the vertical plate 51 is connected to the base 4, the wave plate 52 is arranged on one side of the upper part of the vertical plate 51, the first roller 53 rolls along the wave plate 52, a first rotating shaft 531 is arranged in the first roller 53, and both ends of the first rotating shaft 531 extend from the first roller 53, the lifting drive structure 54 is arranged on the other side of the upper part of the vertical plate 51, and the vibration transmission structure 55 is arranged between the first roller 53 and the carrier plate 2; The step drive mechanism 6 is arranged at the lower end of the base 4, and the synchronous drive mechanism 6 includes a second rotating shaft 61, four transmission components 62 and a rotation drive structure 63. The second rotating shaft 61 is vertically arranged in the middle of the base 4, and the four transmission components 62 are arranged in a ring array around the second rotating shaft 61. The transmission component 62 includes a first transmission shaft 621 and a second transmission shaft 622. The first transmission shaft 621 is fixed to the bottom of the base 4 through a bearing seat, and the second transmission shaft 622 is fixed to the vertical plate 51 through a bearing seat. The first transmission shaft 621 and the second transmission shaft 622 are connected by two bevel gears, and the first transmission shaft 621 and the second rotating shaft 61 are connected by a bevel gear set. The second transmission shaft 622 is connected by a lifting drive structure 54. The four transmission components 62 are used to drive the four lifting and vibration synchronization mechanisms 5 to operate at the same time, and the rotation drive structure 63 is used to drive the second rotating shaft 61 to rotate.

[0041] The staff puts a plurality of test tubes containing biological samples and specifically bound magnetic particle solutions into the test tube holder 1, and then fixes the test tube holder 1 on the carrier plate 2. Then the synchronous drive mechanism 6 works, the rotary drive structure 63 drives the second rotating shaft 61 to rotate, and the second rotating shaft 61 drives the four lifting and vibration synchronous mechanisms 5 to operate through four transmission components 62 respectively. The lifting drive structure 54 drives the first roller 53 to roll from top to bottom along the wave plate 52. During the rolling of the first roller 53 along the wave plate 52, the distance between the first roller 53 and the carrier plate 2 changes repeatedly. When the distance between the first roller 53 and the carrier plate 2 decreases, the vibration transmission structure 55 is compressed. When the distance between the first roller 53 and the carrier plate 2 increases, the compressed vibration transmission structure 55 When released, the vibration transmission structure 55 will generate slight vibration in the process of compression and release. The four vibration transmission structures 55 transmit the vibration to the carrier plate 2 from all sides of the carrier plate 2 at the same time, so that the carrier plate 2 maintains horizontal vibration to prevent the solution in the test tube from spilling. After the first roller 53 rolls to the lower end of the wave plate 52, the biological sample and the specifically bound magnetic particle solution in the test tube are fully mixed, and all the test tubes are located within the working range of the magnetic separation mechanism 3. The continuous rotation of the magnetic separation mechanism 3 allows the magnetic particles in the solution to carry the bound target substance and be adsorbed to the side wall to achieve side wall agglomeration, thereby completing the magnetic separation of the target substance and the non-target substance, so that the solution is evenly mixed and is in the magnetic separation mechanism 3, thereby eliminating the time of transferring the test tube to the magnetic separation mechanism 3.

[0042] Reference Figure 3 , Figure 4 and Figure 5 As shown: the vibration transmission structure 55 includes a connecting plate 551, a first connecting column 552, a compression block 553 and a first spring 554; the connecting plate 551 is connected to the bearing plate 2; there are a plurality of first connecting columns 552, and the plurality of first connecting columns 552 are evenly distributed at both ends of the connecting plate 551, and one end of the first connecting column 552 is connected to the connecting plate 551; there are two compression blocks 553, and the two compression blocks 553 are respectively arranged at both ends of the first rotating shaft 531, and both ends of the compression blocks 553 are plugged into the first connecting column 552; there are a plurality of first springs 554, and the plurality of first springs 554 are respectively sleeved on the plurality of first connecting columns 552, and the two ends of the first spring 554 are respectively abutted against the connecting plate 551 and the compression block 553.

[0043] Since the force is evenly applied on all sides of the supporting plate 2, the supporting plate 2 will not move in the horizontal direction. When the distance between the first roller 53 and the supporting plate 2 decreases, the first roller 53 pushes the compression block 553 to move along the first connecting column 552 toward the connecting plate 551 through the first rotating shaft 531, and the first spring 554 between the compression block 553 and the connecting plate 551 is compressed. When the distance between the first roller 53 and the supporting plate 2 increases, the force exerted by the first roller on the compression block 553 toward the connecting plate 551 decreases, and the first spring 554 releases its elastic potential energy to push the compression block 553 along the first connecting column 552 away from the connecting plate 551. The first spring 554 is repeatedly compressed and released at a fixed frequency, so that the first spring 554 vibrates, and the vibration is transmitted to the supporting plate 2 through the connecting plate 551, thereby realizing repeated vibration of the first spring 554 at a fixed frequency, avoiding the increase of vibration amplitude caused by vibration superposition, and maintaining the vibration frequency of the test tube.

[0044] Reference Figure 3 , Figure 4 and Figure 6 As shown: the lifting drive structure 54 includes a screw 541, a lifting block 542 and two connecting claws 543; the screw 541 is vertically arranged on the vertical plate 51; the middle part of the lifting block 542 is threadedly connected to the screw 541, and the screw 541 rotates to drive the lifting block 542 to move along the axial direction of the screw 541; the two connecting claws 543 are respectively arranged at both ends of the lifting block 542, one end of the connecting claw 543 is connected to the lifting block 542, and the other end of the connecting claw 543 is provided with a strip groove 5431, and the end of the first rotating shaft 531 is slidably arranged in the strip groove 5431.

[0045] When the second rotating shaft 61 rotates, it is evenly transmitted to the four screw rods 541 vertically arranged on the vertical plate 51 through the four transmission components 62. Since the screw rods 541 and the lifting blocks 542 are threadedly connected, the rotation of the screw rods 541 directly causes the lifting blocks 542 to move up and down along the axial direction of the screw rods 541. Since the four screw rods 541 are synchronously rotated through the transmission components 62, the four lifting blocks 542 are also synchronously lifted and lowered accordingly, thereby ensuring that the bearing plate 2 connected to the lifting blocks 542 always remains in a horizontal state during the lifting process. During the lifting process of the lifting block 542, the groove wall of the strip groove 5431 exerts a vertical force on the first rotating shaft 531, ensuring that the first rotating shaft 531 and the first roller 53 connected thereto can be stably lifted and lowered in the vertical direction. At the same time, since the strip groove 5431 has a certain length, the first rotating shaft 531 can move along the strip groove 5431, so that the first roller 53 can generate a horizontal displacement during the rolling process, thereby achieving the coordinated unification of the lifting and rolling of the first roller 53.

[0046] Reference Figure 4 and Figure 7 As shown: the lifting and vibration synchronization mechanism 5 also includes two stabilizing structures 56, and the two stabilizing structures 56 are respectively arranged on both sides of the vertical plate 51, and the stabilizing structure 56 includes a first guide structure 561 and two balancing connecting arms 562; the first guide structure 561 includes a first guide rod 5611 and a first slider 5612, the first guide rod 5611 is arranged parallel to the screw rod 541, and the first slider 5612 is slidably arranged on the first guide rod 5611; the two balancing connecting arms 562 are arranged in parallel, one end of the balancing connecting arm 562 is connected to the end of the first slider 5612, and the two balancing connecting arms 562 are abutted against the upper and lower surfaces of the connecting claw 543, and the other end of the balancing connecting arm 562 is connected to the supporting plate 2.

[0047] The lifting block 542 drives the first roller 53 to rise and fall through the two connecting claws 543, and the first roller 53 drives the supporting plate 2 to rise and fall through the vibration transmission structure 55. Since the force transmission path for driving the supporting plate 2 to rise and fall is long, the force has hysteresis, which easily leads to uneven force on the four sides of the supporting plate 2. By setting two stabilizing structures 56, when the system is in a non-working state, the supporting plate 2 is supported by four vertical plates 51, keeps horizontal and perpendicular to the vertical plates 51, and the two stabilizing structures 56 are respectively fixed on both sides of the vertical plates 51 to ensure the stability of the entire system. The first guide rod 5611 is installed parallel to the screw rod 541, and the first slider 5612 is stationary on the first guide rod 5611 in the initial position. The balancing connecting arm 562 is connected to the first slider 5612 at one end and the supporting plate 2 at the other end. At the same time, the two balancing connecting arms 562 and the upper and lower connecting claws 543 are connected. The surfaces are in close contact. As the screw rod 541 rotates, the lifting block 542 starts to move up and down through the connecting claw 543. Since the connecting claw 543 is connected to the balancing connecting arm 562, the lifting and lowering movement of the connecting claw 543 will be immediately transmitted to the first slider 5612 through the balancing connecting arm 562, so that the first slider 5612 is synchronously lifted and lowered along the first guide rod 5611. During this process, the balancing connecting arm 562 always remains perpendicular to the vertical plate 51. Since the other end of the balancing connecting arm 562 is connected to the supporting plate 2 and remains perpendicular to the vertical plate 51, the supporting plate 2 is not only subjected to the direct force from the lifting block 542 and the vibration transmission structure 55 during the lifting process, but also obtains additional stability support through the balancing connecting arm 562, thereby effectively reducing the hysteresis and unevenness in the force transmission process, so that the four sides of the supporting plate 2 are more evenly stressed.

[0048] Reference Figure 7 and Figure 8As shown: the stabilizing structure 56 also includes a vibration buffer structure 563, which is arranged on the supporting plate 2. The vibration buffer structure 563 is used to connect the balancing connecting arm 562 and the supporting plate 2. The vibration buffer structure 563 includes a second guide rod 5631 and a second slider 5632. The second guide rod 5631 is horizontally arranged, and the second slider 5632 is slidably arranged on the second guide rod 5631. The upper and lower ends of the second slider 5632 are respectively axially connected to the two balancing connecting arms 562.

[0049] If the balancing connecting arm 562 is fixedly connected to the supporting plate 2, the supporting plate 2 and the vertical plate 51 will be rigidly connected, and the vibration transmission structure 55 will be difficult to vibrate the supporting plate 2. By setting a vibration buffer structure 563, the vibration buffer structure 563 is set on the supporting plate 2, and a flexible connection between the balancing connecting arm 562 and the supporting plate 2 is achieved through the combination of the second guide rod 5631 and the second slider 5632. The balancing connecting arm 562 applies a vertical force to the supporting plate 2 through the second slider 5632, ensuring that the supporting plate 2 can maintain balance in the vertical direction and effectively resist external disturbances. When the supporting plate 2 is vibrated, the second slider 5632 can slide freely along the second guide rod 5631, so that the connection point between the balancing connecting arm 562 and the supporting plate 2 can be dynamically adjusted, thereby solving the vibration transmission problem caused by the rigid connection.

[0050] Reference Figure 4 and Fig. 9 As shown: the lifting and vibration synchronization mechanism 5 also includes an ejection structure 57, which is arranged in the middle of the lifting block 542, and the ejection structure 57 includes a first bracket 571, a push rod 572, an arc-shaped top plate 573 and a second spring 574; the first bracket 571 is connected to the lifting block 542; the push rod 572 is slidably connected to the first bracket 571, and the push rod 572 can move along its axial direction; the arc-shaped top plate 573 is arranged at one end of the push rod 572, and the inner concave surface of the arc-shaped top plate 573 abuts against the middle of the first roller 53; the second spring 574 is sleeved on the other end of the push rod 572, and the two ends of the second spring 574 abut against the end of the push rod 572 and the first bracket 571 respectively, and the ejection structure 57 also includes a linear drive unit 575, which is used to apply a force along the axial direction of the push rod 572 to the push rod 572.

[0051] The magnetic separation mechanism 3 adsorbs the target substance carried by the magnetic particles in the solution onto the side wall. After the magnetic separation is completed, the test tube holder 1 needs to be lifted for the next step of operation. The second spring 574 pushes the arc-shaped top plate 573 to fit tightly against the middle of the first roller 53. Due to the concave surface design of the arc-shaped top plate 573, uniform contact between the arc-shaped top plate 573 and the first roller 53 can be ensured. The linear drive unit 575 applies a force along the axis direction of the top rod 572 to the top rod 572, so that the top rod 572 obtains a stable and continuous force. As the arc-shaped top plate 573 moves toward the supporting plate 2 along the axis direction of the top rod 572, the first roller 53 is smoothly pushed away, and the first roller 53 is separated from the wave plate 52, effectively avoiding the vibration caused by the contact between the first roller 53 and the wave plate 52 during the lifting process, thereby preventing the separated target substance from the non-target substance from being re-mixed.

[0052] Reference Fig. 9 and Fig.10 As shown, a plurality of second rollers 5731 are disposed on the concave side of the arc-shaped top plate 573 , and the plurality of second rollers 5731 are all tangent to the first roller 53 .

[0053] The arc-shaped top plate 573 is pressed against the first roller 53 under the action of the second spring 574. When the first roller 53 rolls along the wave plate 52, the arc-shaped top plate 573 will generate resistance to the movement of the first roller 53. By arranging a plurality of second rollers 5731 on the arc-shaped top plate 573, the plurality of second rollers 5731 are kept in a tangent state with the first roller 53. When the first roller 53 rolls along the wave plate 52, the first roller 53 not only rotates itself, but also drives the second rollers 5731 on the arc-shaped top plate 573 to rotate together in a tangential manner, thereby converting the sliding friction between the first roller 53 and the arc-shaped top plate 573 into the rolling friction between the first roller 53 and the second roller 5731. Compared with sliding friction, rolling friction has smaller friction resistance and smoother motion characteristics, thereby making the first roller 53 smoother during rolling, reducing vibration and energy loss caused by friction.

[0054] Reference Figure 3 , Fig.11 and Fig.12As shown: the rotation driving structure 63 includes a gear 631, a rectangular frame 632, a second guide structure 633, a continuous transmission structure 634, a third rotating shaft 635, a transmission connecting rod assembly 636 and a U-shaped frame 638; the gear 631 is connected to the second rotating shaft 61; the rectangular frame 632 is covered on the outside of the gear 631; the second guide structure 633 has at least two, and the two second guide structures 633 are respectively arranged on both sides of the rectangular frame 632. The second guide structure 633 is used to limit the moving direction of the rectangular frame 632. The second guide structure 633 includes a third guide rod 6331 and a third slider 6332. The third guide rod 6331 is parallel to the base 4. The third slider 6332 is slidably arranged on the third guide rod 6331, and the third slider 6332 is connected to the rectangular frame 632; the continuous transmission structure 634 has two, Two continuous transmission structures 634 are symmetrically arranged on both sides of the gear 631, and the continuous transmission structure 634 includes a transmission rack 6341 and a linear driver 6342. The transmission rack 6341 is arranged on one side of the gear 631, and the linear driver 6342 is fixed on one side of the rectangular frame 632. The linear driver 6342 is used to drive the transmission rack 6341 to engage and disengage with the gear 631; the third rotating shaft 635 is vertically arranged at one end of the rectangular frame 632; the transmission connecting rod assembly 636 is arranged between the third rotating shaft 635 and the rectangular frame 632, and the transmission connecting rod assembly 636 is used to convert the rotation of the third rotating shaft 635 into a reciprocating linear motion of the rectangular frame 632 along the guide direction of the second guide structure 633. The two ends of the U-shaped frame 638 are connected to the base 4, and the middle part of the third rotating shaft 635 is connected to the U-shaped frame 638 through a bearing.

[0055] After the first roller 53 moves from the upper end of the wave plate 52 to the lower end of the wave plate 52, if the solution in the test tube is not mixed evenly, the first roller 53 needs to be moved from the lower end of the wave plate 52 to the upper end of the wave plate 52. At this time, the rotation direction of the screw 541 needs to be changed, and the rotation drive structure 63 needs to change the rotation direction of the second rotating shaft 61. By setting the gear 631, the rectangular frame 632, the second guide structure 633, the continuous transmission structure 634, the third rotating shaft 635 and the transmission connecting rod assembly 636, the third rotating shaft 635 is driven to rotate by the motor set on the U-shaped frame 638 through the bevel gear set, and the third rotating shaft 635 rotates through the transmission connecting rod assembly 636 to drive the rectangular frame 632 to reciprocate along the third guide rod 6331. When the rectangular frame 632 is moved by the third guide rod 6331 In the process of moving from one end to the other end, a transmission rack 6341 meshes with the gear 631 under the action of the corresponding linear driver 6342, and the movement of the transmission gear 631 drives the gear 631 to rotate. When the rectangular frame 632 is reset, the aforementioned transmission rack 6341 is separated from the gear 631, and the other transmission rack 6341 meshes with the gear 631. The transmission rack 6341 drives the gear 631 to continue to rotate in the same direction, and the gear 631 drives the second rotating shaft 61 to rotate in one direction. When the second rotating shaft 61 needs to rotate in another direction, the working order of the aforementioned two transmission racks 6341 is exchanged, and the rotation direction of the gear 631 is changed, so that the third rotating shaft 635 rotates in one direction to adjust the rotation direction of the screw 541.

[0056] Reference Fig.11 and Fig.13 As shown: the rotation driving structure 63 also includes a linkage structure 637, which is arranged between the magnetic separation mechanism 3 and the third rotating shaft 635. The linkage structure 637 includes two transmission wheels 6371 and a transmission belt 6372. The two transmission wheels 6371 are respectively connected to the magnetic separation mechanism 3 and the third rotating shaft 635, and the two ends of the transmission belt 6372 are respectively mounted on the two transmission wheels 6371.

[0057] From the above, it can be seen that the rotation direction of the third shaft 635 is fixed by rotating in one direction to adjust the rotation direction of the screw 541. Therefore, the rotation force of the third shaft 635 can be transmitted to the magnetic separation mechanism 3. By setting the linkage structure 637, when the third shaft 635 rotates to drive the carrier plate 2 to rise and fall, the third shaft 635 not only drives the movement of the rectangular frame 632, but also starts to rotate as a part of the linkage structure 637. The third shaft 635 is directly connected to a transmission wheel 6371. When the third shaft 635 rotates, the transmission wheel 6371 also rotates accordingly. The transmission wheel 6371 is connected to another transmission wheel 6371 through a transmission belt 6372 to form a closed transmission loop, so that the other transmission wheel 6371 also starts to rotate, thereby achieving synchronous rotation between the third shaft 635 and the magnetic separation mechanism 3, thereby improving the working efficiency of the overall equipment.

[0058] The method for using a flow pre-treatment device for detecting surface antigens and intracellular proteins comprises the following steps:

[0059] S1, firstly put a plurality of test tubes containing biological samples and specifically bound magnetic particle solutions into a test tube holder 1, and then fix the test tube holder 1 on a carrying plate 2;

[0060] S2, the rotation drive structure 63 drives the second rotating shaft 61 to rotate, and the second rotating shaft 61 drives the four lifting drive structures 54 to operate simultaneously through the four transmission components 62;

[0061] S3, the lifting drive structure 54 drives the first roller 53 to move from top to bottom along the wave plate 52;

[0062] S4, when the first roller 53 rolls along the wave plate 52, the distance between the first roller 53 and the carrying plate 2 changes repeatedly, the vibration transmission structure 55 is repeatedly compressed and released, and the vibration generated by the compression and release is transmitted to the carrying plate 2, and the carrying plate 2 drives the test tube to vibrate;

[0063] S5, after the first roller 53 rolls to the lower end of the wave plate 52, the biological sample in the test tube and the specifically bound magnetic particle solution are fully mixed, and all the test tubes are located in the magnetic separation mechanism 3. The continuous rotation of the magnetic separation mechanism 3 causes the magnetic particles in the solution to carry the bound target substances to be adsorbed to the side wall to achieve side wall aggregation, thereby completing the magnetic separation of the target substance and the non-target substance.

[0064] The above embodiments only express one or several implementation modes of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the present invention. It should be pointed out that, for those of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the attached claims.

Claims

1. A flow pretreatment device for detecting surface antigens and intracellular proteins, comprising a carrier plate (2) for placing a test tube holder (1) and a base (4) for installing a magnetic separation mechanism (3), characterized in that: It also includes four lifting and vibration synchronization mechanisms (5) and a synchronization drive mechanism (6); Four lifting and vibration synchronization mechanisms (5) are respectively arranged at the middle of four sides of the bearing plate (2). The lifting and vibration synchronization mechanism (5) comprises a vertical plate (51), a wave plate (52), a first roller (53), a lifting drive structure (54) and a vibration transmission structure (55). The lower end of the vertical plate (51) is connected to the base (4). The wave plate (52) is arranged on one side of the upper part of the vertical plate (51). The first roller (53) rolls along the wave plate (52). A first rotating shaft (531) is arranged in the first roller (53), and both ends of the first rotating shaft (531) extend out of the first roller (53). The lifting drive structure (54) is arranged on the other side of the upper part of the vertical plate (51). The vibration transmission structure (55) is arranged between the first roller (53) and the bearing plate (2). The synchronous drive mechanism (6) is arranged at the lower end of the base (4), and comprises a second rotating shaft (61), four transmission components (62) and a rotation drive structure (63). The second rotating shaft (61) is vertically arranged in the middle of the base (4), and the four transmission components (62) are arranged in a circular array around the second rotating shaft (61). The four transmission components (62) are used to simultaneously drive the four lifting and vibration synchronous mechanisms (5) to operate, and the rotation drive structure (63) is used to drive the second rotating shaft (61) to rotate.

2. The flow pretreatment device for detecting surface antigens and intracellular proteins according to claim 1, characterized in that: The vibration transmission structure (55) comprises a connecting plate (551), a first connecting column (552), a compression block (553) and a first spring (554); The connecting plate (551) is connected to the carrying plate (2); There are a plurality of first connecting columns (552), and the plurality of first connecting columns (552) are evenly distributed at both ends of the connecting plate (551), and one end of the first connecting column (552) is connected to the connecting plate (551); There are two compression blocks (553), which are respectively arranged at two ends of the first rotating shaft (531), and both ends of the compression blocks (553) are plugged into the first connecting column (552); There are a plurality of first springs (554), and the plurality of first springs (554) are respectively sleeved on a plurality of first connecting pillars (552), and two ends of the first springs (554) are respectively in contact with the connecting plate (551) and the compression block (553).

3. The flow pretreatment device for detecting surface antigens and intracellular proteins according to claim 1, characterized in that: The lifting drive structure (54) comprises a screw rod (541), a lifting block (542) and two connecting claws (543); The screw rod (541) is vertically arranged on the vertical plate (51); The middle portion of the lifting block (542) is threadedly connected to the screw rod (541), and the screw rod (541) rotates to drive the lifting block (542) to move along the axis direction of the screw rod (541); The two connecting claws (543) are respectively arranged at two ends of the lifting block (542); one end of the connecting claw (543) is connected to the lifting block (542); the other end of the connecting claw (543) is provided with a strip groove (5431); and the end of the first rotating shaft (531) is slidably arranged in the strip groove (5431).

4. The flow pretreatment device for detecting surface antigens and intracellular proteins according to claim 1, characterized in that: The lifting and vibration synchronization mechanism (5) further comprises two stabilizing structures (56), the two stabilizing structures (56) being respectively arranged on both sides of the vertical plate (51), and the stabilizing structures (56) comprising a first guide structure (561) and two balancing connecting arms (562); The first guide structure (561) comprises a first guide rod (5611) and a first sliding block (5612); the first guide rod (5611) is arranged parallel to the screw rod (541); and the first sliding block (5612) is slidably arranged on the first guide rod (5611); The two balancing connecting arms (562) are arranged in parallel, one end of the balancing connecting arm (562) is connected to the end of the first sliding block (5612), the two balancing connecting arms (562) are in contact with the upper and lower surfaces of the connecting claw (543), and the other end of the balancing connecting arm (562) is connected to the bearing plate (2).

5. The flow pretreatment device for detecting surface antigens and intracellular proteins according to claim 4, characterized in that: The stabilizing structure (56) further comprises a vibration buffer structure (563), wherein the vibration buffer structure (563) is arranged on the bearing plate (2), and the vibration buffer structure (563) is used to connect the balancing connecting arm (562) and the bearing plate (2).

6. The flow pretreatment device for detecting surface antigens and intracellular proteins according to claim 4, characterized in that: The lifting and vibration synchronization mechanism (5) further comprises an ejection structure (57), the ejection structure (57) being arranged in the middle of the lifting block (542), the ejection structure (57) comprising a first bracket (571), an ejector rod (572), an arc-shaped ejector plate (573) and a second spring (574); The first bracket (571) is connected to the lifting block (542); The push rod (572) is slidably connected to the first bracket (571), and the push rod (572) is capable of moving along its axis direction; The arc-shaped top plate (573) is arranged at one end of the top rod (572), and the inner concave surface of the arc-shaped top plate (573) abuts against the middle part of the first roller (53); The second spring (574) is sleeved on the other end of the push rod (572), and the two ends of the second spring (574) are respectively in contact with the end of the push rod (572) and the first bracket (571).

7. The flow pretreatment device for detecting surface antigens and intracellular proteins according to claim 6, characterized in that: A plurality of second rollers (5731) are arranged on the inner concave side of the arc-shaped top plate (573), and the plurality of second rollers (5731) are all tangent to the first roller (53).

8. The flow pretreatment device for detecting surface antigens and intracellular proteins according to claim 1, characterized in that: The rotary drive structure (63) comprises a gear (631), a rectangular frame (632), a second guide structure (633), a continuous transmission structure (634), a third rotating shaft (635), a transmission connecting rod assembly (636) and a U-shaped frame (638); The gear (631) is connected to the second rotating shaft (61); The rectangular frame (632) is covered on the outside of the gear (631); There are at least two second guide structures (633), the two second guide structures (633) are respectively arranged on both sides of the rectangular frame (632), and the second guide structures (633) are used to limit the moving direction of the rectangular frame (632); There are two continuous transmission structures (634), and the two continuous transmission structures (634) are symmetrically arranged on both sides of the gear (631); The third rotating shaft (635) is vertically arranged at one end of the rectangular frame (632); The transmission connecting rod assembly (636) is arranged between the third rotating shaft (635) and the rectangular frame (632), and the transmission connecting rod assembly (636) is used to convert the rotation of the third rotating shaft (635) into a reciprocating linear motion of the rectangular frame (632) along the guide direction of the second guide structure (633); Both ends of the U-shaped frame (638) are connected to the base (4), and the middle portion of the third rotating shaft (635) is connected to the U-shaped frame (638) via a bearing.

9. The flow pretreatment device for detecting surface antigens and intracellular proteins according to claim 8, characterized in that: The rotary drive structure (63) further comprises a linkage structure (637), wherein the linkage structure (637) is arranged between the magnetic separation mechanism (3) and the third rotating shaft (635).

10. A method for using a flow pretreatment device for detecting surface antigens and intracellular proteins, applied to the flow pretreatment device for detecting surface antigens and intracellular proteins according to any one of claims 1 to 9, characterized in that: The following steps are involved: S1, firstly placing a plurality of test tubes containing biological samples and specifically bound magnetic particle solutions into a test tube holder (1), and then fixing the test tube holder (1) on a carrier plate (2); S2, the rotation drive structure (63) drives the second rotating shaft (61) to rotate, and the second rotating shaft (61) drives the four lifting drive structures (54) to operate simultaneously through the four transmission components (62); S3, the lifting drive structure (54) drives the first roller (53) to move from top to bottom along the corrugated plate (52); S4, when the first roller (53) rolls along the wave plate (52), the distance between the first roller (53) and the support plate (2) changes repeatedly, the vibration transmission structure (55) is repeatedly compressed and released, and the vibration generated by the compression and release is transmitted to the support plate (2), and the support plate (2) drives the test tube to vibrate; S5, after the first roller (53) rolls to the lower end of the wave plate (52), the biological sample in the test tube and the specifically bound magnetic particle solution are fully mixed, and all the test tubes are located in the magnetic separation mechanism (3). The continuous rotation of the magnetic separation mechanism (3) causes the magnetic particles in the solution to carry the bound target substances and be adsorbed to the side wall, thereby achieving side wall aggregation, and completing the magnetic separation of the target substance and the non-target substance.

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