Five-order magnetic separation cleaning method for chemiluminescence immunoassay

Abstract

The invention discloses a five-order magnetic separation cleaning method for chemiluminescence immunoassay, which is realized through a magnetic separation device, and comprises the following steps: under the action of a rotary driving mechanism, a driving disc rotates by a certain angle each time, reaction cups on the driving disc alternately enter a suction and injection position and an avoidance position, and after the driving disc rotates by 360 degrees, the reaction cups are separated from the suction and injection position; the reaction cup completes a complete magnetic washing process once, and before the reaction cup reaches the suction and injection position opposite to the substrate needle, the substrate needle spits the substrate to the water receiving tank at the avoiding position twice. The magnetic separation device is simple in structure, small in size, low in cost and low in failure rate, five-order magnetic separation is adopted, and immune complexes are fully cleaned; the reaction cup clamping holes in the driving disc are distributed at intervals, and gaps among the reaction cup clamping holes can be used for empty injection of substrates, so that the substrates can be injected into the water receiving tank before formal injection of the substrates, and the problem of variation of first hole results of the substrates is solved.

Description

Technical Field

[0001] The invention relates to the technical field of microanalysis detection equipment, in particular to a fifth-order magnetic separation and cleaning method for chemiluminescence immunoassay. Background Art

[0002] Chemiluminescent immunoassay (CLI) is a microanalytical detection technique that combines the high sensitivity of chemical reactions with the high specificity of immunoassays. It is one of the most advantageous quantitative immunoassay methods developed after enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and fluorescent immunoassay (FLIA). Magnetic separation is a key step in CLI. The magnetic separation process involves cleaning the magnetic beads within the reaction vessel. Breaking up the beads facilitates cleaning within the reaction vessel. Common methods for breaking up the beads include mechanical breaking up and magnetic arrangement. Mechanical breaking up is complex, costly, and prone to failure. Magnetic arrangement can both break up and aggregate the beads, but some existing magnetic arrangement structures can cause the beads to aggregate in a dry state for extended periods within the reaction vessel, leading to poor cleaning effectiveness and a high concentration of impurities within the immune complex, affecting experimental results. Some arrangements also require fewer cleaning cycles, failing to completely remove impurities from the immune complex, impacting experimental results. In addition, the existing magnetic separation cleaning method has the following problems during use: the magnetic separation cleaning method has a complex structure, large volume, and occupies a large area; when placing the reaction container, liquid splashing is likely to occur, which is not conducive to cleaning and can easily cause the experimental results to jump; the substrate is heated for a long time to volatilize and partially denature, which affects the results; the substrate mixing mechanism has a relatively complex structure, high cost, and high failure rate; there is no anti-collision trigger alarm function when the washing head descends, resulting in the suction and injection needle being damaged, or the wrong result cannot be predicted and alarmed; crystallization is prone to occur on the drive disk and needs to be cleaned frequently; the liquid flow in the water tank is not smooth, and blockage occurs when crystals accumulate. Summary of the Invention

[0003] In order to solve the above problems, the present invention provides a fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay, which can specifically adopt the following technical solutions: The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay of the present invention is realized by a magnetic separation device. The magnetic separation device includes a base plate, on which a lifting drive mechanism, a rotating drive mechanism and an online mixing mechanism are provided, the lifting drive mechanism is connected to the head washing assembly, the head washing assembly includes an arc-shaped plate, on which five sets of elution needles and one set of liquid-pipetting needles are evenly spaced, and a set of substrate needles is provided at one end of the arc-shaped plate close to the liquid-pipetting needles through a connecting plate, and an anti-collision mechanism is also provided on the arc-shaped plate; the rotating drive mechanism is connected to a driving disk, which is concentrically arranged with the arc-shaped plate, and a magnetic field generator, a water receiving trough and an online mixing mechanism are provided below the driving disk, a plurality of lugs are evenly spaced on the periphery of the driving disk, and each of the lugs is correspondingly provided with a reaction cup clamping hole, and the reaction cup clamping hole has an aspiration position corresponding to the elution needle / liquid-pipetting needle / substrate needle and an avoidance position for avoiding the elution needle / liquid-pipetting needle / substrate needle; The magnetic separation cleaning method comprises: Under the action of the rotary drive mechanism, each time the drive disk rotates a certain angle, the reaction cup on it alternately enters the aspiration position and the avoidance position. After the drive disk rotates 360 degrees, the reaction cup completes a complete magnetic washing process. Before the reaction cup reaches the aspiration position opposite to the substrate needle, the substrate needle spits the substrate into the water receiving tank twice at the avoidance position.

[0004] Preferably, the lifting drive mechanism includes a screw motor arranged on the base plate, the lifting nut of the screw motor is fixedly connected to the arc plate, and vertical guide rods are also provided on both sides of the screw of the screw motor, and the vertical guide rods are connected to the arc plate through linear bearings.

[0005] Preferably, the elution needle is used for aspirating and injecting liquid, and includes a sleeve-type elution needle and a parallel-type elution needle; the elution needle and the aspiration needle are both connected to the arc plate via equi-height screws sleeved with compression springs.

[0006] Preferably, the anti-collision mechanism includes an anti-collision column passing through the arc-shaped plate, a photoelectric sensor is correspondingly arranged above the anti-collision column, and a buffer spring is arranged between the photoelectric sensor and the anti-collision column.

[0007] Preferably, the rotation drive mechanism includes a stepper motor arranged below the base plate, the output shaft of the stepper motor is connected to the central axis of the driving disk through a belt transmission assembly, and the driving disk is spaced apart from the magnetic field generator.

[0008] Preferably, the water receiving trough is an arc-shaped trough corresponding to the position of the arc-shaped plate, which is arranged on the bottom plate, with a drainage hole provided at each end, and the bottom surface is provided with a slope descending from the middle to the two ends.

[0009] Preferably, the magnetic field generator is arranged on the water receiving trough, and includes an inner magnetic bracket and an outer magnetic bracket respectively arranged on the inner and outer sides of the reaction cup, and the inner magnetic bracket and the outer magnetic bracket are both used to set magnetic parts.

[0010] Preferably, the online mixing mechanism is arranged on a bottom plate via a fixed column, and includes a driver and a mixer, wherein an output shaft of the driver is fixedly connected to a rotating shaft, a guide column extending radially is provided on the rotating shaft, the mixer is sleeved on the rotating shaft, a spiral groove for passing the guide column is provided on the side wall of the mixer, a support is provided on the top of the mixer, a support groove for eccentrically placing a reaction cup is provided on the top of the support, and a counterweight is provided at the bottom of the mixer; The driver works in the following manner during the lifting and mixing process: In the first step, the output shaft of the driver enters the first uniform acceleration stage, which is used to make the mixer rise. During this process, the guide post moves from the top starting end of the spiral groove to the bottom ending end of the spiral groove; In the second step, the output shaft of the driver enters the second uniform acceleration stage, which is used to keep the mixer rotating at the highest position. During this process, the speed of the output shaft is always greater than the speed in the first uniform acceleration stage. In the third step, the output shaft of the driver first rotates at a uniform speed and then enters a uniform deceleration stage until the speed reaches zero. During this process, the mixer descends and the guide column reaches the top starting end of the spiral groove from the bottom end end of the spiral groove.

[0011] Preferably, an exhaust hole is provided on the side wall of the mixer, and a drainage hole communicating with the supporting groove is provided on the side wall of the support.

[0012] Preferably, the bottom plate is also provided with a splash-proof mechanism, which includes a mounting seat fixedly arranged on the bottom plate, a reaction cup accommodating groove is provided at the top center of the mounting seat, and buffer cotton sheets connected to the side walls of the reaction cup are provided on both sides of the top of the mounting seat, and the buffer cotton sheets are fixed by an L-shaped pressing sheet, and the L-shaped pressing sheet is fixed to the side walls of the mounting seat by bolts.

[0013] The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay provided by the present invention is realized by a magnetic separation device with simple structure, small size, low cost and low failure rate. It adopts fifth-order magnetic separation, cleans the immune complex cleanly and has low background. The reaction cup card placement holes on the driving disk are spaced apart, and the gaps therein can be emptied with substrate. Therefore, before formally adding substrate, the substrate can be injected into the water receiving tank, solving the problem of variation in the results of the first hole of the substrate. The driving disk adopts a hollow structure, and the droplets hung by the injection needle can all fall into the water receiving tank, avoiding the problem of crystallization requiring manual cleaning. The water tank is provided with a slope and double-sided water outlets to avoid crystallization blockage and ensure smooth water discharge. The elution needle adopts a sleeve-type elution needle and a parallel elution needle integrated into one. Therefore, after the aspiration is completed, Liquid injection can reduce the dry state time of the magnetic bead immune complex, isolate the liquid medium between the magnetic bead immune complexes, avoid strong magnetic acceleration causing the sticky cup to be unable to be cleaned, or agglomerate the magnetic bead immune complex into a mass that cannot be mixed, resulting in inaccurate experimental results; it has a head washing and anti-collision function, the reaction container position is abnormal or the turntable rotates abnormally, and the head washing and anti-collision can avoid the suction injection needle from being damaged and the predicted result may have a problem alarm function; a flexible buffer cotton sheet is provided under the cup inlet, and the reaction container is placed in the downward pressure process, first contacting the flexible material, which can avoid visible splashing and the impact on the experimental results; the substrate mixing selects a single motor to rotate and automatically rise, eccentric mixing, and stop automatically descending to give way. It has a good mixing effect and does not take up much space. It can be seen that the magnetic separation and cleaning method described in the present invention can effectively improve the accuracy of the reaction results. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a structural schematic diagram of the magnetic separation device of the present invention.

[0015] Figure 2 yes Figure 1 Schematic diagram of the structure of the washing head component.

[0016] Figure 3 yes Figure 1 Schematic diagram of the structure of other components after removing the shampoo component.

[0017] Figure 4 yes Figure 3 sectional view.

[0018] Figure 5 yes Figure 3 Schematic diagram of the exploded structure of the central water tank and magnetic field generator.

[0019] Figure 6 yes Figure 4 Enlarged view of the magnetic field generator part.

[0020] Figure 7 yes Figure 4 Enlarged view of the splash protection mechanism.

[0021] Figure 8 yes Figure 1 Schematic diagram of the structure of the online mixing mechanism.

[0022] Figure 9 make Figure 8 sectional view.

[0023] Figure 10 yes Figure 8 Schematic diagram of the three-dimensional structure of the mixer and support.

[0024] Figure 11 1 is a diagram showing the relationship between the rotational speed and time of the driver in the embodiment. DETAILED DESCRIPTION

[0025] The following describes an embodiment of the present invention in detail with reference to the accompanying drawings. This embodiment is implemented based on the technical solution of the present invention, and provides a detailed implementation method and a specific working process. However, the protection scope of the present invention is not limited to the following embodiment.

[0026] like Figure 1-11 As shown, the fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay described in the present invention is implemented by a magnetic separation device, which is used to separate the immune complex, unreacted substances and other substances in the reaction container (such as the reaction cup M), thereby removing other substances other than the immune complex. The immune complex carries magnetic particles, and the immune complex with magnetic particles moves in a direction under the action of the magnetic field. The aspiration and injection are performed on the liquid in the reaction container, thereby achieving the effect of cleaning the immune complex.

[0027] The magnetic separation device comprises a base plate 1, a drive disk 2, a shampoo assembly, a lifting drive mechanism, a rotary drive mechanism, an online mixing mechanism, a splash prevention mechanism, an anti-collision mechanism, a magnetic field generator, and a water receiving trough. The lifting drive mechanism, rotary drive mechanism, online mixing mechanism, splash prevention mechanism, and water receiving trough are connected to the base plate 1, while the drive disk 2 is connected to the rotary drive mechanism. The shampoo assembly is located above the drive disk 2 and is connected to the lifting drive mechanism.

[0028] Specifically, the head washing assembly includes a curved plate 31, on which are evenly spaced five sets of elution needles 32 and one set of aspiration needles 33. A set of substrate needles 34 is provided at one end of the curved plate 31, near the aspiration needles 33, via a connecting plate. The central axes of the elution needles 32, aspiration needles 33, and substrate needles 34 are cocircular.

[0029] The elution needle 32 is used for aspiration and injection, and includes a sleeve elution needle and four parallel elution needles (see Figure 2The sleeve-type elution needle functions as a cleaning and aspiration needle, preventing BF carryover. Aspiration needle 33 is used for aspiration, achieving fifth-order magnetic separation; substrate needle 34 is used for substrate addition. Both the elution needle 32 and aspiration needle 33 are connected to the curved plate 31 via a spherical screw 36, which is sheathed with a compression spring 35. This provides a cushioning effect when the needle tip hits the bottom, protecting it from damage.

[0030] Furthermore, an anti-collision mechanism is provided on the curved plate 31. The anti-collision mechanism includes an anti-collision post 41 extending through the curved plate 31. A photoelectric sensor 42 is provided above the anti-collision post 41. The photoelectric sensor 42 is fixedly connected to the curved plate 31, and a buffer spring 43 is provided between the photoelectric sensor 42 and the anti-collision post 41. When the anti-collision post 41 is hit, it pushes up and triggers the photoelectric sensor 42, thereby protecting the elution needle 32 and the aspiration needle 33.

[0031] To perform operations such as adding liquid, the shampoo assembly is connected to a lifting drive mechanism. In this embodiment, the lifting drive mechanism includes a screw motor 51 disposed on the base plate 1. The lifting nut of the screw motor 51 is fixedly connected to the curved plate 31. Vertical guide rods 52 are also provided on both sides of the screw of the screw motor 51. The vertical guide rods 52 are connected to the curved plate 31 via linear bearings 53.

[0032] Below the shampoo assembly is a drive disk 2 connected to a rotary drive mechanism. In this embodiment, the rotary drive mechanism includes a stepper motor 61 disposed below the base plate 1, and the output shaft of the stepper motor 61 is connected to the central axis of the drive disk 2 via a belt drive assembly 62.

[0033] The drive disk 2 is a hollow structure, concentrically arranged with the curved plate 31 (i.e., the center of the drive disk 2 coincides with the central axes of the elution needle 32, the aspiration needle 33, and the substrate needle 34). Multiple lugs 21 are evenly spaced around the periphery of the drive disk 2, each of which corresponds to a cuvette receiving hole 22. As the drive disk 2 rotates, each cuvette receiving hole 22 has a corresponding aspiration position for the elution needle 32, the aspiration needle 33, or the substrate needle 34, and a clearance position that avoids the elution needle 32, the aspiration needle 33, or the substrate needle 34. The above-mentioned avoidance position refers to the position of the elution needle 32 / liquid aspiration needle 33 / substrate needle 34 corresponding to the hollow space between the two adjacent reaction cup clamping holes 22, which can be used as a reserved position for empty substrate injection. Before the reaction cup M of the present invention is officially filled with substrate, it runs to the avoidance position twice to perform substrate empty injection, and the substrate is directly discharged into the water receiving tank 7 below, thereby solving the problem of variation in the results of the first substrate hole; secondly, the droplets hung by the injection needle can fall more into the water receiving tank 7, reducing the problem of crystallization on the drive disk 2 and the need for manual cleaning.

[0034] Below the drive disk 2 are a magnetic field generator 8 and a water receiving trough 7, arranged in sequence. The drive disk 2 and magnetic field generator 8 are spaced apart. The water receiving trough 7 is an arc-shaped trough corresponding to the position of the arc-shaped plate 31 (i.e., the tips of the elution needle 32, the liquid aspiration needle 33, and the substrate needle 34 correspond to the arc centerline of the water receiving trough 7). It is fixedly mounted on the base plate 1, with a drainage hole at each end, and the bottom surface has a slope that descends from the center to the ends. This structure ensures smooth water discharge from the water receiving trough 7 and prevents crystallization and clogging. The magnetic field generator 8 is mounted on the water receiving trough and includes an inner magnetic bracket 81 and an outer magnetic bracket 82, respectively positioned on the inner and outer sides of the reaction cup M (i.e., positioned on the inner and outer sides of the reaction cup receiving hole 22). Both the inner and outer magnetic brackets 81 and 82 are used to mount a magnetic element 83.

[0035] In this embodiment, the magnetic members 83 are installed so that the distance between the top of the magnetic member 83 and the bottom of the reaction cup M is 6.85 mm. Specifically, the N-level of the first three magnetic members 83 in the inner magnetic member holder 81 faces the reaction cup M, the S-level of the magnetic members 83 in positions 4 and 5 faces the reaction cup M, and the S-level of the magnetic members 83 in positions 8, 9, 12, and 13 faces the reaction cup M. The S-level of the first three magnetic members 83 in the outer magnetic member holder 82 faces the reaction cup M, the S-level of the magnetic members 83 in positions 6 and 7 faces the reaction cup M, and the S-level of the magnetic members 83 in positions 10 and 11 faces the reaction cup M. The arrangement of the magnetic members 83 in this manner ensures better alignment of the magnetic beads in the reaction cup M, freeing impurities in the immune complex in the liquid medium, and ensuring cleaner cleaning of the immune complex.

[0036] When the driving disk 2 rotates under the action of the rotary driving mechanism, the reaction cup M thereon moves between the outer magnetic component support 82 and the inner magnetic component support 81 , and the magnetic component 83 provides a stable magnetic field for the reaction cup M.

[0037] To prevent splashing caused by collisions when the gripper places the cuvette M, a splash prevention mechanism is provided on the base plate 1. This splash prevention mechanism comprises a mounting base 91 fixed to the base plate 1. A cuvette receiving slot 92 is provided at the center of the top of the mounting base 91. Buffering pads 93 are provided on both sides of the top of the mounting base 91 to contact the side walls of the cuvette M. These cushioning pads 93 are secured by L-shaped pressing pads 94, which are secured to the side walls of the mounting base 91 by bolts 95.

[0038] When the cuvette M is placed on the drive disk 2 by the gripping device, the cuvette placement hole 22 is aligned with the cuvette receiving groove 92. When the cuvette M passes through the cushioning cotton sheet 93, the cushioning cotton sheet 93 has a certain resistance to the cuvette M, and the cuvette M cannot perform free fall. This prevents the cuvette M from colliding with the drive disk 2 due to rapid fall, and prevents the liquid in the cuvette M from splashing, thereby playing a splash-proof role.

[0039] In order to make the liquid in the reaction cup M more evenly mixed, an online mixing mechanism is also installed on the bottom plate 1. The above-mentioned online mixing mechanism includes a driver and a mixer.

[0040] In this embodiment, the driver is a unidirectional motor 1001. Of course, a forward / reversible motor can also be used as the driver. The output shaft of the unidirectional motor 1001 is fixedly connected to the rotating shaft 1002 via a screw 1003, and the two rotate synchronously. A radially extending guide post 1004 is also fixed to the rotating shaft 1002, and the guide post 1004 is used to connect to the mixer 1005.

[0041] The mixer 1005 is made of wear-resistant Iglidur® J engineering plastic and is mounted on the rotating shaft 1002. Its sidewalls are provided with spiral grooves 1006 for guide posts 1004 to pass through. When the unidirectional motor 1001 rotates and there is a speed difference between the rotating shaft 1002 and the mixer 1005, the mixer 1005 rises or falls along the rotating shaft 1002, meaning that the guide posts 1004 slide upward or downward along the spiral grooves 1006. Because the clearance between the rotating shaft 1002 and the mixer 1005 is relatively small, vent holes 1007 are provided in the sidewalls of the mixer 1005 to ensure smooth relative movement between the rotating shaft 1002 and the mixer 1005. Secondly, a continuous ridge 1008 is provided on the spiral channel 1006, which is connected to the guide post 1004. The contact surface width of the ridge 1008 and the guide post 1004 is less than the wall thickness of the mixer 1005. This is used to reduce the contact area between the guide post 1004 and the spiral channel 1006, reduce friction, and ensure smooth relative movement between the rotating shaft 1002 and the mixer 1005. Thirdly, the bottom terminal end of the spiral channel 1006 is arranged horizontally (i.e., a horizontal section is provided), and the entrance of the horizontal section is provided with a boss 1009 that reduces the channel diameter to ensure that the mixer 1005 does not fall when it rises to the highest position. Finally, a counterweight 1010 is provided at the bottom of the mixer 1005. The counterweight 1010 is a counterweight ring arranged coaxially with the mixer 1005, which is used to lower the center of gravity of the mixer 1005 and ensure the operational stability of the mixer 1005.

[0042] A support 1011 is provided on top of the mixer 1005. The support 1011 has an upper-opening support groove 1012, with the central axis of the support groove 1012 parallel to the central axis of the rotating shaft 1002. This means that the support groove 1012 is eccentric. When the mixer 1005 rotates the support 1011, the sample to be tested within the eccentric support groove 1012 is effectively mixed. A drainage hole 1013 is provided on the sidewall of the support 1011, communicating with the support groove 1012. This prevents accidental spillage of the sample from flowing into the support groove 1012 and forming a pool.

[0043] In addition, an upwardly extending fixing column 1014 is respectively installed at the four corners of the top of the housing of the unidirectional motor 1001 for fixing the device to the online analysis equipment.

[0044] During the lifting and mixing process, the rotating shaft 1002 keeps accelerating to drive the mixer 1005 to rotate. The operating speed of the mixer 1005 is always lower than the operating speed of the rotating shaft 1002, thereby ensuring the stable operation of the mixer 1005. The unidirectional motor 1001 has the following three speed control stages: In the first step, the output shaft of the unidirectional motor 1001 enters the first uniform acceleration stage, which is used to make the mixer 1005 rise. During this process, the guide post 1004 moves from the top starting end of the spiral channel 1006 to the bottom ending end of the spiral channel 1006; In the second step, the output shaft of the unidirectional motor 1001 enters the second uniform acceleration stage, which is used to keep the mixer 1005 rotating at the highest position. During this process, the speed of the output shaft of the unidirectional motor 1001 is always greater than the speed in the first uniform acceleration stage. In the third step, the output shaft of the unidirectional motor 1001 first rotates at a uniform speed and then enters a uniform deceleration stage until the speed reaches zero. During this process, the mixer 1005 descends and the guide column 1004 reaches the top starting end of the spiral groove 1006 from the bottom ending end of the spiral groove 1006.

[0045] The working process of the above-mentioned online mixing mechanism is described in detail below: The first uniform acceleration stage (0-0.2 seconds): the rotating shaft 1002 rotates clockwise at an accelerated speed under the action of the unidirectional motor 1001, and the mixer 1005 rotates relative to the rotating shaft 1002 under the action of static inertia. During the relative rotation of the mixer 1005 and the rotating shaft 1002, the guide column 1004 on the rotating shaft 1002 provides a force in the direction of rotation. This force acts on the inclined surface of the spiral groove 1006 of the mixer 1005 and is decomposed into a force in the direction of rotation and a force perpendicular to the direction of rotation and upward. The force in the direction of rotation causes the mixer 1005 to accelerate along with the rotating shaft 1002, and the force perpendicular to the direction of rotation and upward causes the mixer 1005 to overcome its own gravity and the gravity of the counterweight 1010 and rise.

[0046] At this time, if the unidirectional motor 1001 is still maintained in the first uniform acceleration state, due to the inertia of the mixer 1005, the mixer 1005 will collide with the guide column 1004 at the moment it rises to the highest position. After the collision, the guide column 1004 will break away from the horizontal section at the bottom end of the spiral groove 1006 and eventually fall, resulting in a failure to lift.

[0047] Therefore, when the guide post 1004 reaches the bottom end of the spiral groove 1006 , the unidirectional motor 1001 will continue to accelerate and enter the second uniform acceleration stage.

[0048] The second uniform acceleration stage (0.2-3.7 seconds): the output shaft of the unidirectional motor 1001 is Figure 4 The second section of the six-point curve shown is accelerated to overcome the collision between the mixer 1005 and the guide post 1004 caused by the inertia of the mixer 1005. The acceleration force of the second section controlled by the six-point curve is greater than the force of the collision between the mixer 1005 and the guide post 1004, so that the rotating shaft 1002 is continuously accelerated to drive the mixer 1005 to rotate. That is, the running speed of the mixer 1005 is always lower than the running speed of the rotating shaft 1002, thereby achieving stable operation of the mixer 1005.

[0049] The unidirectional motor 1001 is driven by pulses. After receiving a pulse sequence, the motor 1001 starts rotating. However, the pulse frequency received by the motor 1001 fluctuates, causing the motor 1001 to operate at an uneven speed. During this uneven speed, when the speed of the rotating shaft 1002 is slower than that of the mixer 1005, the mixer 1005 will fall, resulting in mixing failure. In the present invention, the speed of the motor 1001 is controlled according to the second segment of the six-point curve. This allows the motor 1001 to overcome the fluctuations in its speed caused by the pulse frequency fluctuations, allowing the rotating shaft 1002 to continuously accelerate and drive the mixer 1005 to rotate. This ensures stable operation of the mixer 1005, even if the mixer 1005's speed is always slower than that of the rotating shaft 1002.

[0050] When the mixer 1005 is running stably, the guide post 1004 is in the horizontal groove at the bottom of the mixer 1005. At this time, the force exerted by the guide post 1004 on the mixer 1005 is a force along the rotation direction and a force perpendicular to the rotation direction and downward. Due to the boss 1009 provided at the entrance of the horizontal section of the spiral groove 1006, the guide post 1004 is subjected to increased resistance to the separation of the horizontal section of the spiral groove 1006, thereby further ensuring that the mixer 1005 will not fall when rotating at the top.

[0051] Uniform speed buffering stage (3.7-4 seconds): When the mixing is completed, the output shaft of the unidirectional motor 1001 is Figure 4 The third section of the six-point curve shown maintains a uniform rotation speed to buffer the subsequent deceleration movement and protect the unidirectional motor 1001 from damage.

[0052] Uniform deceleration stage (4-4.2 seconds): In this stage, the output shaft of the unidirectional motor 1001 is Figure 4The fourth section of the six-point curve shown enters a uniform deceleration stage until the speed reaches zero.

[0053] During the above-mentioned uniform speed buffering stage and uniform deceleration stage, the mixer 1005 loses the driving effect of the rotating shaft 1002. When the rotating shaft 1002 stops rotating, the mixer 1005 continues to rotate clockwise under the action of inertia. Since the rotating shaft 1002 and the guide post 1004 are stationary, the guide post 1004 will be separated from the horizontal groove of the mixer 1005. The mixer 1005 drops to the lowest position under the action of gravity of the mixer 1005 and the counterweight block 1010, and finally reaches a stationary state.

[0054] The above-mentioned online mixing mechanism is well-suited for use in online analytical equipment. It features a simple structure, compact size, easy operation, stable operation, and a low failure rate. It eliminates the need for additional locking devices or magnetic mechanisms to prevent the mixer from falling. Simply by controlling the driver's speed, the mixer's stable rise and high-position rotation are ensured, even when the mixer is constantly accelerating during the mixing process. When mixing is complete, the driver simply slows down. This online mixing mechanism improves the overall space utilization of the equipment and reduces its footprint.

[0055] The magnetic separation cleaning method of the present invention comprises: The cuvette M is placed into the cuvette receiving slot 92 of the splash prevention mechanism using a gripper mechanism. Once the cuvette M is placed, the rotary drive mechanism rotates the drive disk 2 20 times, each rotation 18°. After the cuvette M rotates 360° along with the drive disk 2, a complete magnetic washing process is completed. During these 20 rotations of the drive disk 2, the cuvette holder 22, where the cuvette M is located, alternates between the aspiration and injection position and the avoidance position, with the cuvette M positioned in the 10-time aspiration and injection position and the 10-time avoidance position, respectively. The aspiration and injection position refers to the position where the cuvette holder 22 corresponds to the elution needle 32, the aspiration needle 33, and the substrate needle 34. In this position, aspiration and injection of liquid from the cuvette M can be completed. The avoidance position refers to the position where the cuvette holder 2 clears the elution needle 32, the aspiration needle 33, and the substrate needle 34. When the drive disk 2 is in this position, the substrate needle 34 can directly discharge liquid into the water receiving tank 7 below. Among them, before the first reaction cup M for batch testing on the driving disk 2 reaches the aspiration position opposite to the substrate needle 34, the substrate needle 34 spits the substrate directly into the water receiving tank 7 twice at the avoidance position, thereby solving the influence of evaporation of the first hole substrate on the experimental results.

[0056] The magnetic separation and cleaning process is described in detail below through specific embodiments: 1) The cuvette M is placed on the drive disk 2 by the gripper device. After the drive disk 2 rotates 36 degrees, the first cuvette M in the batch test (hereinafter referred to as the cuvette M) comes to the second aspiration position. The immune complex with magnetic particles in the cuvette M is attracted to the two sides of the cuvette M by the magnetic elements 83 on both sides. 2) After the drive disk 2 rotates another 36°, the reaction cup M reaches the third aspiration position. The immune complex with magnetic particles in the reaction cup M is attracted to both sides of the reaction cup M by the magnetic members 83 on both sides. The washing head assembly slowly descends under the drive of the screw motor 51, and the sleeve elution needle (i.e., the first elution needle 32) aspirates the liquid in the reaction cup M. After the liquid is aspirated, the washing head assembly rises, and the sleeve elution needle simultaneously injects liquid into the reaction cup M. Specifically, the immune complexes with magnetic particles are adsorbed to both sides of the reaction cup M by the magnetic members 83 on both sides. Unreacted substances and impurities are free in the liquid. The sleeve-type elution needle aspirates the unreacted substances and impurities along with the liquid during aspiration, while the immune complexes with magnetic particles remain on both sides of the reaction cup M. After the liquid in the reaction cup M is aspirated, the liquid is quickly injected into the reaction cup M, thereby preventing the immune complexes with magnetic particles from aggregating. 3) After the drive disk 2 rotates another 36°, the cuvette M reaches the fourth aspiration position. The immune complex containing the magnetic particles in the cuvette M is attracted to the inner side of the cuvette M by the magnetic member 83 on the inner magnetic member support 81. The wash head assembly slowly descends under the drive of the lead screw motor 51, and the parallel elution needle (i.e., the second elution needle 32) aspirates the liquid in the cuvette M. After the liquid is aspirated, the wash head assembly rises, and the parallel elution needle simultaneously injects liquid into the cuvette M. 4) After the drive disk 2 rotates another 36°, the reaction cup M reaches the fifth aspiration position. The immune complex containing magnetic particles in the reaction cup M is attracted to the outside of the reaction cup M by the magnetic member 83 on the external magnetic member support 82. The washing head assembly slowly descends under the drive of the screw motor 51, and the parallel elution needle (i.e., the third elution needle 32) aspirates the liquid in the reaction cup M. After the liquid is aspirated, the washing head assembly rises, and the parallel elution needle simultaneously injects liquid into the reaction cup M. 5) After the drive disk 2 rotates another 36°, the cuvette M reaches the sixth aspiration position. The immune complex containing the magnetic particles in the cuvette M is attracted to the inner side of the cuvette M by the magnetic member 83 on the inner magnetic member support 81. The wash head assembly slowly descends under the drive of the lead screw motor 51, and the parallel elution needle (i.e., the fourth elution needle 32) aspirates the liquid in the cuvette M. After the liquid is aspirated, the wash head assembly rises, and the parallel elution needle simultaneously injects liquid into the cuvette M. 6) After the drive disk 2 rotates another 36°, the cuvette M reaches the seventh aspiration position. The immune complex containing the magnetic particles in the cuvette M is attracted to the outside of the cuvette M by the magnetic member 83 on the external magnetic member support 82. The wash head assembly slowly descends under the drive of the lead screw motor 51, and the parallel elution needle (i.e., the fifth elution needle 32) aspirates the liquid in the cuvette M. After the liquid is aspirated, the wash head assembly rises, and the parallel elution needle simultaneously injects liquid into the cuvette M. 7) After the drive disk 2 continues to rotate 18°, the cuvette M reaches the seventh avoidance position, and the substrate needle 34 discharges liquid toward the water tank 7 below for the first time; 8) After the drive disk 2 continues to rotate 18°, the reaction cup M reaches the eighth aspiration position. The immune complex containing magnetic particles in the reaction cup M is attracted to the inner side of the reaction cup M by the magnetic member 83 on the inner magnetic member bracket 81. The washing head assembly slowly descends under the drive of the screw motor 51, and the aspiration needle 33 aspirates the liquid in the reaction cup M. After the liquid is aspirated, the washing head assembly rises, and the immune complex containing magnetic particles in the reaction cup M is now cleaned. 9) After the drive disk 2 rotates another 18°, the cuvette M reaches the eighth avoidance position, and the substrate needle 34 discharges liquid toward the water tank 7 below for the second time. 10) After the drive disk 2 rotates another 18°, the reaction cup M reaches the ninth aspiration position. The wash head assembly slowly descends under the drive of the screw motor 51. When the wash head assembly reaches the lowest point, the substrate needle 34 injects liquid into the reaction cup M. After the liquid injection is completed, the wash head assembly rises under the drive of the screw motor 51 and returns to the original position. 11) After the drive disk 2 continues to rotate 36°, the cuvette M reaches the tenth aspiration position. After the cuvette M reaches this position, the rotating shaft 1002 of the online mixing mechanism begins to rotate, and the mixer 1005 rises to mix the immune complex and substrate in the cuvette M. After the mixing is completed, the mixer 1005 automatically falls; 12) After the drive disk 2 continues to rotate 36°, the reaction cup M reaches the first aspiration position, and the entire cleaning process is completed.

[0057] In the above process, the immune complex with magnetic particles in the reaction cup M swims back and forth five times between the inner and outer walls of the reaction cup M under the action of the magnetic field of the magnetic member 83 , thereby completely cleaning the immune complex.

[0058] It should be noted that, in the description of the present invention, terms indicating orientation or positional relationships such as “front”, “rear”, “left”, “right”, “vertical”, “horizontal”, “inside”, and “outside” are based on the orientation or positional relationships shown in the accompanying drawings and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention.

Claims

1. A fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay, implemented by a magnetic separation device, characterized in that: The magnetic separation device includes a base plate, on which a lifting drive mechanism, a rotating drive mechanism and an online mixing mechanism are provided, the lifting drive mechanism is connected to the head washing assembly, the head washing assembly includes an arc-shaped plate, on which five sets of elution needles and one set of liquid-pipetting needles are evenly spaced, and a set of substrate needles is provided at one end of the arc-shaped plate close to the liquid-pipetting needles through a connecting plate, and an anti-collision mechanism is also provided on the arc-shaped plate; the rotating drive mechanism is connected to a driving disk, which is concentrically arranged with the arc-shaped plate, and a magnetic field generator, a water receiving trough and an online mixing mechanism are provided below the driving disk, a plurality of lugs are evenly spaced on the periphery of the driving disk, and each of the lugs is correspondingly provided with a reaction cup clamping hole, and the reaction cup clamping hole has an aspiration position corresponding to the elution needle / liquid-pipetting needle / substrate needle and an avoidance position for avoiding the elution needle / liquid-pipetting needle / substrate needle; The magnetic separation cleaning method comprises: Under the action of the rotary drive mechanism, each time the drive disk rotates a certain angle, the reaction cup on it alternately enters the aspiration position and the avoidance position. After the drive disk rotates 360 degrees, the reaction cup completes a complete magnetic washing process. Before the reaction cup reaches the aspiration position opposite to the substrate needle, the substrate needle spits the substrate into the water receiving tank twice at the avoidance position.

2. The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay according to claim 1, characterized in that: The lifting drive mechanism includes a screw motor arranged on the base plate, the lifting nut of the screw motor is fixedly connected to the arc plate, vertical guide rods are also arranged on both sides of the screw of the screw motor, and the vertical guide rods are connected to the arc plate through linear bearings.

3. The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay according to claim 1, characterized in that: The elution needle is used for aspirating and injecting liquid, and includes a sleeve-type elution needle and a parallel-type elution needle; the elution needle and the aspiration needle are both connected to the arc plate via equal-height screws sleeved with compression springs.

4. The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay according to claim 1, characterized in that: The anti-collision mechanism includes an anti-collision column passing through the arc plate, a photoelectric sensor is correspondingly arranged above the anti-collision column, and a buffer spring is arranged between the photoelectric sensor and the anti-collision column.

5. The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay according to claim 1, characterized in that: The rotary drive mechanism includes a stepper motor arranged below the base plate. The output shaft of the stepper motor is connected to the central axis of the driving disk through a belt transmission assembly, and the driving disk is spaced apart from the magnetic field generator.

6. The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay according to claim 1, characterized in that: The water receiving trough is an arc-shaped trough corresponding to the position of the arc-shaped plate, which is arranged on the bottom plate, with a drainage hole provided at each end, and the bottom surface is provided with a slope descending from the middle to the two ends.

7. The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay according to claim 1, characterized in that: The magnetic field generator is arranged on the water receiving trough, and includes an inner magnetic bracket and an outer magnetic bracket respectively arranged on the inner and outer sides of the reaction cup, and the inner magnetic bracket and the outer magnetic bracket are both used to set magnetic parts.

8. The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay according to claim 1, characterized in that: The online mixing mechanism is arranged on the bottom plate via a fixed column, and includes a driver and a mixer. The output shaft of the driver is fixedly connected to the rotating shaft. The rotating shaft is provided with a guide column extending in the radial direction. The mixer is sleeved on the rotating shaft. The side wall of the mixer is provided with a spiral groove for passing the guide column. The top of the mixer is provided with a support. The top of the support is provided with a support groove for eccentrically placing a reaction cup. The bottom of the mixer is provided with a counterweight block. The driver works in the following manner during the lifting and mixing process: In the first step, the output shaft of the driver enters the first uniform acceleration stage, which is used to make the mixer rise. During this process, the guide post moves from the top starting end of the spiral groove to the bottom ending end of the spiral groove; In the second step, the output shaft of the driver enters the second uniform acceleration stage, which is used to keep the mixer rotating at the highest position. During this process, the speed of the output shaft is always greater than the speed in the first uniform acceleration stage. In the third step, the output shaft of the driver first rotates at a uniform speed and then enters a uniform deceleration stage until the speed reaches zero. During this process, the mixer descends and the guide column reaches the top starting end of the spiral groove from the bottom end end of the spiral groove.

9. The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay according to claim 8, characterized in that: An exhaust hole is provided on the side wall of the mixer, and a drainage hole communicating with the supporting groove is provided on the side wall of the support.

10. The fifth-order magnetic separation and cleaning method for chemiluminescent immunoassay according to claim 1, characterized in that: The bottom plate is also provided with a splash-proof mechanism, which includes a mounting seat fixedly arranged on the bottom plate, a reaction cup accommodating groove is provided at the top center of the mounting seat, and buffer cotton sheets connected to the side walls of the reaction cup are provided on both sides of the top of the mounting seat. The buffer cotton sheets are fixed by an L-shaped pressing sheet, and the L-shaped pressing sheet is fixed to the side walls of the mounting seat by bolts.

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