Multi-channel full-automatic intelligent protein interaction test equipment

Abstract

The invention relates to the technical field of biological information, in particular to multi-channel full-automatic intelligent protein interaction test equipment which comprises a test platform and a cover arranged on the test platform, a protein reaction device, a liquid relief device and a liquid storage device are arranged on the test platform, and the protein reaction device comprises a reaction table rotationally connected to the test platform; the reaction table is provided with a plurality of accommodating holes for mounting sample tubes, and a magnetic part for magnetically separating magnetic beads in the sample tubes is arranged in the reaction table; the pipetting device is used for adding and sucking liquid into the sample tube, and comprises a pipetting gun and a displacement mechanism for driving the pipetting gun to horizontally and vertically move; the liquid storage device is used for storing various experimental liquids at low temperature and comprises a low-temperature liquid storage tank and a cover plate arranged on the low-temperature liquid storage tank, and a plurality of independent grooves are formed in the low-temperature liquid storage tank and used for independently placing liquid boxes of different types of liquids. By implementing the scheme, the labor cost is reduced, and the multi-channel full-automatic intelligent protein interaction test equipment which is low in cost and has an automatic function is provided.

Description

Technical Field

[0001] The present invention relates to the field of bioinformatics, and in particular to a multi-channel fully automatic intelligent protein interaction test device. Background Art

[0002] Co-IP experiment is a co-immunoprecipitation experiment, which is mainly used by researchers to study the interaction between proteins and determine which proteins bind to the target protein to form a complex under physiological conditions in the cell. The characteristics of this experiment are:

[0003] 1. The experimental process is relatively cumbersome, including but not limited to: magnetic bead pretreatment (multiple resuspension of magnetic beads), antibody binding (antibody dilution, gentle resuspension of magnetic beads and antibody and overnight low-temperature inversion), precipitation reaction (multiple resuspension, overnight low-temperature inversion after protein binding, magnetic separation, multiple resuspension, high-temperature heating, magnetic separation, etc.);

[0004] 2. The content of each experiment is different because humans have tens of thousands of proteins and a huge antibody library. Even for the same protein, different designs are required when studying its role in different pathways or diseases. In addition, each experiment requires a new control group (negative control, positive control) to be redesigned for the new target.

[0005] 3. Time-consuming: One experiment takes researchers nearly 2 days.

[0006] At present, the general situation of scientific researchers conducting this experiment is manual operation. The reasons for this situation are: 1) Due to the second experimental feature of the Co-IP experiment mentioned above, that is, the content of each experiment is different and the control group of each experiment is different, the relevant parameters cannot be set uniformly; 2) Since the experiment involves many variables and cumbersome operation steps, and this field belongs to the experimental field with a small application market, its R&D cost is relatively high. Therefore, the experiment currently relies on scientific researchers to perform specific operations, and the experiment takes a long time, nearly 2 days. If the scientific research personnel use most of their energy in tedious experimental operations, it will greatly compress their time and energy for innovative research, which is not conducive to scientific and technological progress.

[0007] To address the above issues, there is an urgent need to develop a low-cost and automated protein interaction test equipment to free researchers from tedious scientific research operations and conduct more creative scientific research. Summary of the Invention

[0008] The present invention aims to provide a low-cost, multi-channel, fully automatic, intelligent protein interaction test device with automation functions.

[0009] To achieve the above-mentioned objectives, the present invention adopts the following technical solutions: a multi-channel fully automatic intelligent protein interaction test equipment, comprising a test platform and a cover body arranged on the test platform, the test platform is provided with a protein reaction device, a pipetting device and a liquid storage device, the protein reaction device comprises a reaction table rotatably connected to the test platform, the reaction table is provided with a plurality of accommodating holes for installing sample tubes, and the reaction table is provided with a magnetic part for magnetically separating magnetic beads in the sample tubes; the pipetting device is used to add liquid to and aspirate liquid from the sample tube, comprising a pipette gun and a displacement mechanism for driving the pipette gun to move horizontally and vertically; the liquid storage device is used to store various experimental liquids at low temperatures, comprising a low-temperature liquid storage tank and a cover plate arranged on the low-temperature liquid storage tank, the low-temperature liquid storage tank is provided with a plurality of independent grooves for independently placing liquid boxes of different types of liquids.

[0010] Preferably, as an improvement, a cooling module and a heating module are provided at each receiving hole corresponding to the reaction table, which are used to cool and heat the sample tubes. The top surface of the reaction table is rotatably connected to a flip plate, which is used to simultaneously open and close the tube openings of each sample tube. A top heating module is provided on the flip plate.

[0011] Preferably, as an improvement, a tube cover plate is rotatably connected to the tube mouth of the sample tube, and a cover protrusion for sealing the tube mouth is provided on the tube cover plate. A cover hole is also provided on the tube cover plate. When the tube mouth is sealed by the tube cover plate, the cover hole is misaligned with the end face of the tube mouth of the sample tube.

[0012] Preferably, as an improvement, the free end of the flip cover plate is provided with a cover opening mechanism corresponding to the cover opening hole, the cover opening mechanism includes a connecting shaft and a cover opening member, the connecting shaft is vertically arranged on the lower surface of the flip cover plate, one end of the cover opening member is rotatably connected to the side wall of the connecting shaft, the gravity of the cover opening member is greater than the rotational friction between the cover opening member and the connecting shaft; when the cover opening member is folded, the maximum transverse width of the cover opening mechanism is smaller than the diameter of the cover opening hole, and when the cover opening member is opened, the maximum transverse width of the cover opening mechanism is greater than the diameter of the cover opening hole.

[0013] Preferably, as an improvement, the displacement mechanism includes a horizontal displacement unit and a vertical displacement unit, the horizontal displacement unit includes two parallel first slide rails and a second slide rail sliding on the two first slide rails, a fixed block is slidably connected to the second slide rail, the vertical displacement unit is connected to the fixed block, the pipette is provided on the vertical displacement unit, and the vertical displacement unit is used to drive the pipette to move vertically.

[0014] Preferably, as an improvement, it also includes a gun tip processing mechanism, which includes a gun tip replacement unit and a gun tip recovery unit. The gun tip recovery unit is used to collect discarded gun tips of the pipette, and includes a waste channel and a collection box. The collection box is slidably connected to the side wall of the experimental platform, and the waste channel is tilted in the experimental platform and connected to the collection box; the gun tip replacement unit includes a gun tip box placement area and a camera, and the camera is used to photograph the gun tip position in the gun tip box. The pipette moves to a designated position according to the gun tip position information and inserts the corresponding gun tip.

[0015] Preferably, as an improvement, the grooves are divided into a plurality of groove groups, wherein each groove in at least one groove group corresponds to placing various liquid boxes used in protein interaction experiments.

[0016] Preferably, as an improvement, the grooves in the groove group for placing the magnetic bead suspension are interconnected to form a magnetic bead liquid processing tank, a vortex oscillator is provided in the magnetic bead liquid processing tank, and a vibration frame of the vortex oscillator is provided with a plurality of placement holes for placing magnetic bead liquid tubes in parallel, and an opening cover plate is rotatably connected to the vibration frame, and the opening cover plate is used to simultaneously open and close the tube openings of each magnetic bead liquid tube, the structure of the opening cover plate is the same as that of the flip cover plate, and the structure of the magnetic bead liquid tube is the same as that of the sample tube.

[0017] Preferably, as an improvement, it further includes a robotic arm, a marking device and a sample tube box placement area, the robotic arm is used to clamp the sample tube to a specified position, and the marking device is used to mark the sample tube.

[0018] The principles and advantages of this solution are:

[0019] 1. Freeing up researchers' attention: This solution eliminates the need for researchers to participate in the experiment. They simply place the sample tubes on the reaction table before the experiment and check that the cryogenic liquid reservoirs contain sufficient experimental liquids. They can then wait two days after the experiment is complete before analyzing the results. This frees researchers from tedious experimental operations, allowing them to devote more time and energy to innovative research projects and produce more valuable results.

[0020] 2. Simple structure and compact size: This solution uses the ingenious design of the flip plate structure to enable the flip plate to achieve four functions at the same time: 1) It can realize batch opening and closing of each sample tube without generating excess consumables; at the same time, the control process is simple, and only the rotation of the flip plate is required to drive the tube cover to open and seal the sample tube mouth. 2) The flip plate can press the sample tube to prevent the sample tube from being thrown out during the flipping and mixing process. 3) During the flipping process, the flip plate can always press the cover bulge and the sample tube to keep the two in a sealed state, further ensuring that the risk of liquid spillage is avoided. 4) Since the sample tube will have residual liquid on the cover bulge after the flipping and mixing step, especially in the washing step, the suspension needs to be heated at high temperature. At this time, a large amount of liquid will condense on the cover bulge. During the opening process, the liquid on the cover bulge is very likely to splash into the adjacent sample tube, resulting in inaccurate experimental results. Therefore, this solution uses a flip cover plate and directly sets a heating module inside it to heat the cover convex of the sample tube to keep the cover convex dry. In this way, no liquid splashing will occur when the cover is opened, ensuring the accuracy of the experimental results.

[0021] Therefore, through the ingenious design of the structure, one component can achieve multiple functions at the same time, so there is no need to configure corresponding mechanisms separately for each function: opening the cover, clamping, sealing, and preventing liquid splashing, making the overall equipment simpler and more compact, and at the same time lower in cost.

[0022] 3. Diversified experimental selectivity: Incorporating various experimental scenarios, a more diverse experimental process is provided. For example, during the experiment, you can choose whether to retain the liquid at a certain stage for subsequent analysis; based on experimental accuracy, you can choose whether to transfer the reaction liquid to a new sample tube during the washing stage; and in control experiments, there is more selectivity for experimental liquids, etc. In this way, the diverse experimental needs of researchers can be met. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a structural diagram of Example 1.

[0024] Figure 2 Schematic diagram of the protein reaction device.

[0025] Figure 3 This is a schematic diagram of the improved sample tube structure of the present invention.

[0026] Figure 4 A cross-sectional view of the reaction table.

[0027] Figure 5 for Figure 4 A partial enlarged view of middle A.

[0028] Figure 6 for Figure 4 Cross-sectional view of the middle reaction station with the cover opened.

[0029] Figure 7 Schematic diagram of the structure of the pipetting control mechanism.

[0030] Figure 8 for Figure 7 A partial enlarged view of B.

[0031] Figure 9 This is a structural diagram of Example 2. DETAILED DESCRIPTION

[0032] The following is further described in detail through specific implementation methods:

[0033] The reference numerals in the drawings of the specification include:

[0034] Experimental platform 1, protein reaction device 2, support frame 21, reaction table 22, receiving hole 23, magnetic part 24, thermoelectric semiconductor cooling plate 25, flip cover plate 26, cover opening mechanism 27, connecting shaft 271, cover opening member 272, limit plate 28, weight reduction hole 29,

[0035] Liquid transfer device 3, fixed block 31, screw nut mechanism 32, range adjustment motor 33, control head 34, lower horizontal axis 35, liquid transfer gun 36, control button 37, gun head ejection button 38, connecting plate 39, control box 391,

[0036] Liquid storage device 4, groove 41, magnetic bead liquid processing tank 42,

[0037] Gun tip processing mechanism 5, waste channel 51, collection box 52, gun tip box 53,

[0038] Sample tube 6, tube cover 61, cover protrusion 62, cover opening hole 63, limit protrusion 64, positioning guide strip 65,

[0039] Waste liquid pool 7, robotic arm 8, marking device 9, sample tube box placement area 10.

[0040] Example 1

[0041] Basically as attached Figures 1-8As shown: A multi-channel fully automatic intelligent protein interaction test equipment, including a test platform and a cover body arranged on the experimental platform 1, the test platform is provided with a protein reaction device 2, a pipetting device 3, a liquid storage device 4, a gun tip processing mechanism 5 and a waste liquid pool 7, and the waste liquid pool 7 is located below the protein reaction device 2. The protein reaction device 2 is used to perform magnetic separation, flip mixing, high-temperature heating, low-temperature refrigeration and other treatments on the liquid in the sample tube 6; the liquid storage device 4 is used to store various liquids used in the protein interaction test at low temperature, including at least magnetic bead suspension A, washing buffer B, antibody diluent C, protein lysis solution D, and protein loading buffer E. The pipetting device 3 is used to add various liquids in the liquid storage device 4 to the sample tube 6 in stages, and to suck out the liquid in the sample tube 6. The gun tip processing mechanism 5 is used to replace the gun tip of the pipette 36.

[0042] like Figure 2-Figure 6 As shown, the protein reaction device 2 includes a magnetic member 24 and a support frame 21 provided on the experimental platform 1. A reaction table 22 is rotatably provided on the support frame 21. The reaction table 22 is driven by a speed-regulating motor. The speed-regulating motor is fixedly mounted on the support frame 21, and its output shaft is clamped to the reaction table 22. By using a micro speed-regulating motor, the experimenter can adjust the speed of the reaction table 22 according to different needs to achieve the desired experimental effect. The reaction table 22 is provided with a plurality of receiving holes 23 for accommodating sample tubes 6. Each sample tube 6 is used for experiments with different control groups. The number of receiving holes 23 can be 4, 8, 12, etc., and the axes of each receiving hole 23 are located in the same plane. This solution takes four receiving holes 23 as an example.

[0043] When the number of channels (accommodating holes 23) is large, multiple protein reaction devices 2 can be designed. For example, if there are 10 channels, two 5-channel protein reaction devices 2 can be designed accordingly. The purpose of this design is to take into account that, on the one hand, if there are too many channels on the same protein reaction device 2, the pipette 36 will take a certain amount of time to add liquid from the first to the last one, which may affect the reaction effect of the previous channels.

[0044] The magnetic piece 24 can be installed on the magnetic frame and externally installed on the experimental platform 1. By changing the position of the magnetic piece 24, it can be controlled whether it adsorbs the magnetic beads in the sample tube 6. In addition, the magnetic piece 24 can also be vertically embedded in the reaction table 22 and corresponds to the side wall of each receiving hole 23. The magnetic piece 24 is an electromagnet. This solution adopts the latter method. This method has a high degree of integration and makes the entire structure compact and simple. At the same time, it can reduce the magnetic frame and the related structures that drive the movement of the magnetic frame, and avoid interference with other components during its movement. In addition, by using an electromagnet as the magnetic piece 24, in addition to controlling the working state of the electromagnet by turning the power on and off, the magnitude of its magnetic force can also be controlled by changing the current to be suitable for different types of magnetic beads.

[0045] In addition, in this solution, the electromagnet is vertically located on the bottom side wall of the sample tube 6. Its purpose is to adsorb the magnetic beads on the side wall of the sample tube 6 rather than the bottom of the tube, so as to avoid the risk of sucking away the magnetic beads when the pipette tip 36 is inserted into the bottom of the sample tube 6 to absorb the liquid.

[0046] Since, in the antibody binding step of the experiment, the liquid in the sample tube 6 needs to be flipped overnight with the reaction table 22 under low temperature conditions (usually at 4°C), it needs to be refrigerated. At the beginning of the design, constant temperature refrigeration was considered in the experimental chamber, because in addition to the liquid in the sample tube 6, there are various other reaction liquids stored on the experimental platform 1 that also need to be stored at low temperatures, but it was later discovered that this method will cause water mist and even condensed water droplets to form inside the equipment, especially on the pipette gun 36, and there is a high possibility of contamination risk during the pipetting process. Therefore, it was later considered to set a refrigeration module at each receiving hole 23 on the reaction table 22, which not only avoids the generation of water droplets in the equipment, but also saves energy and reduces unnecessary low-temperature consumption in the equipment.

[0047] At the same time, in the washing step in the final stage of the experiment, the suspension needs to be heated at a high temperature (usually at 95°C). In order to avoid the traditional cumbersome process, the sample tube 6 needs to be removed from the reaction table 22 and placed in boiling water for heating. After heating, it needs to be placed on the reaction table 22 for magnetic separation. Therefore, a heating module is set on the reaction table 22 corresponding to each receiving hole 23.

[0048] The heating module and the cooling module can be respectively provided with a heating layer and a cooling layer. However, considering that the space of the reaction table 22 is limited and it needs to be in a flipped state for a long time, the reaction table 22 as a whole needs to be light and compact. After comprehensive consideration, a thermoelectric semiconductor cooling plate 25 with both cooling and heating functions is adopted. It can switch between cooling and heating functions and adjust the temperature by simply changing the direction and size of the current. At the same time, the thermoelectric semiconductor cooling plate 25 has a small structure and can meet the setting space of the corresponding single sample tube 6. The installation position of the thermoelectric semiconductor cooling plate 25 is staggered with the position of the magnetic member 24. A plurality of weight-reducing holes 29 are provided on the reaction table 22, some of which are connected to the thermoelectric semiconductor cooling plate 25. In this way, the weight of the entire reaction table 22 can be reduced and the thermoelectric semiconductor cooling plate 25 is conducive to its own heat dissipation when in a heated state.

[0049] Since in the entire magnetic bead protein interaction experiment, it is necessary to continuously open the cover of the sample tube 6 for aspiration and addition of liquid, and then close the cover of the sample tube 6 in time for magnetic separation or flipping and mixing. At the same time, since in the experiments of the same batch, each sample tube 6 is a control experiment, the sample tubes 6 cannot be cross-flowed, that is, each sample tube 6 must ensure good airtightness, and at the same time, the completion of the current batch of experiments cannot affect the next experiment. Therefore, how to design a sample tube 6 cover that can effectively avoid the risk of mutual contamination and can automatically and uniformly open and close the cover of each sample tube 6 is a very important and critical link.

[0050] Initially, a method was considered for sample tubes 6 with threaded caps that were screwed using two opposing racks. While this method could open all sample tubes 6 at once, it presented several challenges. First, a mechanism for gripping and releasing the sample tubes 6 was required on the reaction table 22 to prevent them from falling during the turning process. Second, the rack-tightening mechanism was complex, requiring a drive mechanism to operate the racks, a lifting mechanism for the cap to spirally ascend or descend, and a translation mechanism to horizontally move the entire rack-tightening mechanism or the cap from above the sample tubes 6. Consequently, the overall structure was complex, involved numerous operational steps, and was costly.

[0051] Then, the method of using a flip cover 26 was considered, in which a cover for sealing each sample tube 6 was directly set on the flip cover 26. This method only requires driving the flip cover 26 to rotate, and its structure and operation method are extremely simple. However, the problem is that when the next batch of protein interaction experiments is carried out, the residual substances on the cover will affect the results of the next experiment. Later, it was considered to stick a disposable sealing film on the flip cover 26, and set multiple covers on the sealing film corresponding to each sample tube 6. In this way, after each experiment is completed, the sealing film is torn off and a new sealing film is to be stuck. However, the problem that may exist in this method is that the accuracy of its position cannot be ensured by manual sticking, resulting in the problem of loose sealing of the sample tube 6 during the experiment and leakage of reagents. In response to this problem, it was later considered to adopt an automated sticking method, in which the sealing film is set into a roll and stored in the experimental chamber. When sticking, it is necessary to consider tearing off the adhesive film on the sealing film, then cutting the sealing film of appropriate length and sending it to the designated position, and then accurately sticking the sealing film by the sticking mechanism with the assistance of the positioning mechanism. Although this method is feasible, it involves additional consumables: sealing film; at the same time, the additional mechanisms involved are too complicated, and the precision requirements required for coordination in the middle are too high, making the overall cost too high, which is not conducive to the original design intention of making the equipment compact and affordable.

[0052] Therefore, after continuous analysis and optimization of the structure, the following structure is finally designed:

[0053] First, if Figure 3As shown, the sample tube 6 is improved: a tube cover 61 is pivotally connected to the tube mouth of the sample tube 6. Tube cover 61 is provided with a sealing protrusion 62 for sealing the tube mouth. Protrusion 62 can be solid or hollow, but this embodiment adopts a hollow design to save materials and costs. The perimeter wall of protrusion 62, located three-quarters of the way away from tube cover 61, is designed with an inclined surface, facilitating easy insertion of protrusion 62 into the sample tube 6. Tube cover 61 also has a cover opening 63. When tube cover 61 is sealed, cover opening 63 is offset from the end face of the tube mouth of the sample tube 6. The primary improvement over conventional sample tubes 6 lies in the simple addition of cover opening 63 to tube cover 61.

[0054] Secondly: Figure 4-Figure 6 As shown, a flap plate 26 is rotatably connected to the top surface of the reaction table 22 via a bearing. This flap plate 26 is used to simultaneously open and close the nozzles of each sample tube 6. In this embodiment, the flap plate 26 is driven by a servo motor fixed to the reaction table 22, with its output shaft connected to the flap plate 26. A lid opening mechanism 27 is vertically mounted on the left end of the flap plate 26, corresponding to the position of the lid opening hole 63. The lid opening mechanism 27 comprises a connecting shaft 271 and two lid opening members 272. The connecting shaft 271 is vertically mounted on the bottom surface of the flap plate 26. The lower ends of the lid opening members 272 are hinged to the lower end of the connecting shaft 271 via a pin. In this embodiment, the lower ends of the lid opening members 272 are U-shaped and rotatably connected to the connecting shaft 271 via a pin. The gravity of the lid-opening member 272 is greater than the rotational friction between the lid-opening member 272 and the connecting shaft 271. This causes the upper end of the lid-opening member 272 to automatically expand outward under its own weight, without the action of any external force. When the two lid-opening members 272 are retracted, the maximum lateral width of the lid-opening mechanism 27 is smaller than the diameter of the lid-opening hole 63, allowing the lid-opening mechanism 27 to be smoothly inserted into the lid-opening hole 63 of the sample tube 6. When the lid-opening members 272 are extended, the maximum lateral width of the lid-opening mechanism 27 is greater than the diameter of the lid-opening hole 63, allowing the lid-opening member 272 to contact the bottom surface of the flip cover 26 during the opening process, directly driving the flip cover 26 to rotate upward until the lid is opened.

[0055] The lower end of the connecting shaft 271 is integrally formed with a limit plate 28. A limit groove 41 is defined in the center of the limit plate 28. The connection between the cover opening member 272 and the rotating shaft is located within this limit groove 41. The contact surface between the limit groove 41 and the cover opening member 272 is an inclined surface. This design not only limits the expansion stroke of the cover opening member 272, but also effectively supports the cover opening member 272 during the opening process, thereby extending the service life of the cover opening mechanism 27.

[0056] Considering that after the sample tube 6 has gone through the flipping and mixing step, liquid will remain on the cover bulge 62. Especially in the washing step, the suspension needs to be heated at high temperature. At this time, a large amount of liquid will condense on the cover bulge 62. During the opening process, the liquid on the cover bulge 62 is very likely to splash into the adjacent sample tube 6, thereby causing inaccurate experimental results.

[0057] To address this issue, this solution incorporates a heating module within the flip cover 26. This heating module can utilize an electric heating wire. Once the sample tube 6 is flipped and mixed, the cover ridge 62 is heated to evaporate any condensed water vapor or water droplets thereon. This ensures a dry surface and prevents liquid from splashing when the cover is opened.

[0058] The liquid storage device 4 is used to store various experimental liquids at low temperatures, including a low-temperature liquid storage tank and a cover provided on the low-temperature liquid storage tank. When the reaction table 22 is turned over overnight, or the device is not in the experimental state, the cover is in a closed state to prevent the cold air in the low-temperature liquid storage tank from leaking out. The low-temperature liquid storage tank is provided with a plurality of independent grooves 41 for independently placing liquid boxes of different types of liquids. Considering that when doing multiple groups of control experiments, the antibody diluent C and protein lysis solution D used in each control group are different, and in order to facilitate researchers to have more selectivity in experiments, as shown in the attached figure, Figure 1 As shown, the grooves 41 are divided into four groups, one row per group, and the number of grooves 41 groups corresponds to the number of channels in the device. Each groove 41 in at least one group is used to place various liquid cartridges used in protein interaction experiments. In the remaining three groups, in addition to placing different types of antibody diluents and protein lysis buffers, the other grooves 41 can optionally be used to place different types of liquids of the same type.

[0059] Wherein, the groove for placing magnetic bead suspension A in the groove group is interconnected to form magnetic bead liquid treatment tank 42, magnetic bead liquid treatment tank 42 is provided with vortex oscillator, the vibration frame of vortex oscillator is provided with a plurality of placement holes for placing magnetic bead liquid pipes side by side, placement hole quantity is identical with groove group quantity, is four, for placing different models of magnetic bead suspension.Rotation is connected with open cover plate on the vibration frame, open cover plate is used for opening and closing the mouth of pipe of each magnetic bead liquid pipe simultaneously, the structure and connection mode of open cover plate are identical with flip cover plate 26, and magnetic bead liquid pipe structure is identical with sample tube 6, does not repeat them here.So, when experiment starts, first vortex oscillator vibrates magnetic bead suspension A, to evenly disperse magnetic beads.

[0060] like Figure 1As shown, the pipetting device 3 is used to add liquid to and absorb liquid from the sample tube 6, including a pipette gun 36 and a displacement mechanism and a pipetting control mechanism that drives the pipette gun 36 to move horizontally and vertically. The displacement mechanism includes a horizontal displacement unit and a vertical displacement unit. The horizontal displacement unit includes two parallel first slide rails. The first slide rails are fixed above the experimental platform 1. A second slide rail is slidably connected to the two first slide rails. A fixed block 31 is slidably connected to the second slide rail. The vertical displacement unit is connected to the fixed block 31. The vertical displacement unit is a screw nut mechanism 32. At least two different types of pipette guns are installed on the fixed block for calling pipette guns of different ranges according to different pipetting requirements. The displacement mechanism can adopt an electric control slide rail, and the movement route of the pipette gun can be controlled by the control system of the electric control slide rail.

[0061] like Figure 7 、 Figure 8 As shown, the pipetting control mechanism is used to control the pipette gun 36 to aspirate, discharge, withdraw the gun head, and adjust the aspiration range. The pipetting control mechanism includes a control box 391, the two sides of which are respectively threadedly connected to the nut seat of the screw nut mechanism and the pipette gun 36. A control unit is vertically slidably connected to the control box 391, and the control unit can swing left and right. The end point of the left swing is directly above the gun head withdrawal button 38 of the pipette gun 36, and the end point of the right swing is directly above the control button 37 of the pipette gun 36. The control unit includes a range adjustment motor 33 and two control heads 34 (hereinafter referred to as the upper control head 34 and the lower control head 34) coaxially mounted at the output end of the range adjustment motor 33. A gap is left between the two control heads 34. The control heads 34 and the control buttons 37 are both gear structures, and the two can mesh with each other. An L-shaped control frame is fixedly connected to the range adjustment motor 33, and the lower horizontal axis 35 of the control frame passes horizontally through the gap space and protrudes from the right side. As a further improvement, a rotary drum is rotatably sleeved on the lower horizontal shaft 35 of the control frame, so as to reduce the wear of the lower horizontal shaft 35 and the control button 37 during the range adjustment process.

[0062] The control unit is driven by a driving mechanism, which is installed in a control box 391 and includes a servo motor and a control cylinder. The control cylinder is fixedly connected to the output end of the servo motor. A connecting plate 39 is laterally provided at the output end of the control cylinder, and the range adjustment motor 33 is fixed to the bottom of the connecting plate 39.

[0063] The pipetting control process is as follows:

[0064] 1. Adjusting the range of the pipette 36: To increase the pipetting volume, the drive mechanism moves the control head 34 to the right until the upper control head 34 engages with the control button 37 and the lower horizontal shaft 35 contacts the bottom of the control button 37. At this point, the range adjustment motor 33 rotates the control button 37, and the control cylinder cooperates to drive the control button 37 upward until the target range is reached. Conversely, to decrease the pipetting volume, the lower control head 34 engages with the control button 37 and the lower horizontal shaft 35 contacts the top surface of the control button 37.

[0065] 2. Liquid aspiration and liquid discharge: move the control head 34 to the top of the control button 37 and push the control button 37 downward to the first stop point. Release the control button 37 to reset it, completing the liquid aspiration. Push the control button 37 downward again to the second stop point, completing the liquid discharge, and release the control head 34.

[0066] 3. To unload the gun head, the servo motor drives the control head 34 to rotate above the gun head unloading button 38. The control cylinder then drives the control head 34 to push the gun head unloading button 38 downward until the gun head is unloaded and the control head 34 is reset upward.

[0067] Furthermore, a camera is installed at the bottom of the fixed block. It uses captured images to identify the specific position of the pipette and then controls the pipette control mechanism to perform the relevant actions. If the image shows that the pipette is stopped at the waste channel, the pipette control mechanism performs the aforementioned tip removal action. If the image shows that the pipette is stopped at the groove for storing wash buffer B1, the vertical displacement unit drives the pipette downward until the bottom of the tip is below the liquid surface, and the pipette control mechanism performs the aforementioned liquid aspiration action.

[0068] Because this experiment involves a wide variety of liquids and numerous channels, the pipette 36 needs to have its tip replaced each time before adding a different liquid and before aspirating the liquid from different sample tubes 6 to avoid the risk of cross-contamination. Therefore, a tip handling mechanism 5 is designed. The tip handling mechanism 5 includes a tip replacement unit and a tip recovery unit. The tip recovery unit is used to collect discarded tips replaced by the pipette 36 and includes a waste channel 51 and a collection box 52. The collection box 52 is slidably connected to the side wall of the experimental platform. A handle is provided on the right side wall of the collection box 52, allowing researchers to easily remove the collection box 52 and discard the discarded tips. The waste channel 51 is tiltedly arranged in the experimental platform 1 and is connected to the collection box 52; the tip replacement unit includes a tip box 53 placement area and a camera, and the tip placement area includes at least two for placing tips of different sizes and models; the camera is used to photograph the position of the tip in the tip box 53, and the pipette 36 moves to the designated position according to the tip position information and inserts the corresponding tip, thereby completing the replacement of the tip of the pipette 36.

[0069] To prevent the lid-opening mechanism 27 from pulling the sample tube 6 out during the opening process, a horizontal stopper protrusion 64 is provided on the outer wall of the sample tube 6, and a stopper groove is provided within the receiving hole 23 to engage with the stopper protrusion 64, thereby securing the two. Furthermore, to ensure precise alignment between the lid-opening hole 63 of the sample tube 6 and the lid-opening mechanism 27, allowing the lid-opening mechanism 27 to be accurately inserted into the lid-opening hole 63, at least one positioning guide bar 65 is integrally formed on the outer wall of the sample tube 6. This positioning guide bar 65 extends axially along the sample tube 6, and a vertical positioning guide groove is provided within the receiving hole 23 for the positioning guide bar 65 to be inserted into. This allows the experimenter to insert the positioning guide bar 65 into the corresponding positioning guide groove when placing the sample tube 6, ensuring a one-to-one alignment between the lid-opening hole 63 and the lid-opening mechanism 27. Furthermore, the top of the positioning guide groove is flared, with the opening facing upward, to simplify alignment.

[0070] In addition, in order to meet the more diverse experimental demands of experimenters, sample tubes 6 of different capacities will be used. Therefore, reaction tables 22 with various specifications of accommodating holes 23 are designed for sample tubes 6 of different specifications, and the reaction tables 22 can be detachably connected to the support frame 21, such as by clipping, bolting, etc.

[0071] Example 2

[0072] Considering that during the experiment, researchers may retain the liquid at a certain stage for subsequent analysis, such as the liquid after magnetic separation after antigen-antibody binding in the precipitation reaction needs to be retained for analysis; in addition, for experiments with high precision requirements, such as in the washing stage, after washing and resuspension multiple times, the reaction liquid needs to be transferred to a new sample tube 6 to continue the experiment. Therefore, in order to meet the researchers' requirements for more experimental selectivity, the following optimization is made:

[0073] like Figure 9 As shown, a robotic arm 8, a marker 9 and a sample tube box placement area 10 are installed on the experimental platform 1. The robotic arm 8 has six degrees of freedom and is used to clamp the sample tube 6 to a specified position. The marker 9 is used to mark the sample tube 6. The robotic arm 8 and the marker 9 are fixedly installed between the sample tube box placement area 10 and the protein reaction device 2 to shorten the movement route and time of the robotic arm 8.

[0074] An electromagnet rod is installed in the connecting shaft 271 , and the cover opening member 272 is made of iron material or an iron sheet is provided on the cover opening member 272 .

[0075] In this way, when it is necessary to retain the liquid for analysis, the pipette gun 36 absorbs the liquid in the four channels into the new sample tubes 6 placed in the corresponding sample tube box placement area 10. After each new sample tube 6 is completed or all are completed, the robotic arm 8 labels the new sample tubes 6 corresponding to each channel. For example, if the channels on the reaction table 22 are numbered 1, 2, 3, and 4 from left to right, the liquids absorbed from the new sample tubes 6 corresponding to the channels are labeled 1-1, 2-1, 3-1, and 4-1 respectively. This makes it easier for scientific researchers to clearly distinguish the corresponding relationship between the new sample tubes 6 and the original sample tubes 6.

[0076] When the experimental accuracy is high and the sample tube 6 needs to be replaced, the electromagnet rod in the connecting shaft 271 is energized and magnetized, thereby attracting the cover opening member 272 and closing it. The flip cover plate 26 flips upward to separate the cover opening mechanism 27 from the tube cover plate 61 of the sample tube 6. At this time, the sample tube 6 is in a free state, and the robot arm 8 removes the sample tube 6 from the reaction table 22 and transfers the liquid in the tube to a new sample tube 6. The new sample tube 6 is then marked with the corresponding channel number and placed on the reaction table 22. Another transfer method is: if there is an idle protein reaction device 2, the robot arm 8 first grabs a new sample tube 6 from the sample tube 6 box, marks it, and then places it on the protein reaction device 2. The robot arm 8 then transfers the liquid in the original sample tube 6 to the new sample tube 6 to continue the experiment.

[0077] Furthermore, as an optimization, in order to avoid the antibody being eluted during the experiment, areas for placing cross-linking agent (DSS), DMSO, biological buffer (Tris-HCl), BSA, and a blank container (for configuring the cross-linking agent) are also provided in the low-temperature liquid storage tank. In the actual experiment, after the antibody is bound to the magnetic beads, DSS and DMSO are added to the blank container through a pipette 36 to dissolve DSS in DMSO, and then the dissolved DSS is added to the sample tube 6 and rotated and incubated for 30-60 minutes. Tris-HCl is then added and incubated for 15 minutes to neutralize the unreacted DSS. Finally, it is blocked with a buffer containing 1% BSA to reduce nonspecific binding. After thoroughly washing the magnetic beads, continue to the routine.

[0078] The specific operation method is as follows:

[0079] First, the vortex oscillator is started to evenly disperse the magnetic beads in the magnetic bead suspension A. The cover plate is then driven to flip and open the cover of the magnetic bead liquid tube. The pipette gun 36 is controlled to move to the magnetic bead suspension A1 to aspirate and inject the liquid into the first sample tube 6 from the left to the right. The aspiration and injection are then repeated until the last sample tube 6 is completed. The pipette gun 36 is controlled to move to the top of the waste channel 51 to remove the tip. Then, based on the position of the remaining tips in the tip box 53 captured by the camera, the pipette gun 36 is controlled to move to the designated tip and insert it. Then, wash buffer B1 is respectively aspirated into the four sample tubes 6 for washing. After washing is completed, the electromagnet is controlled to conduct magnetic separation on the liquid in the sample tube 6, and the separated liquid is aspirated and discharged. Then, wash buffer B1 is re-added and the above steps are repeated three times. Finally, 200ul of wash buffer B1 is injected into each sample tube 6 for resuspending and standby use, thereby obtaining liquid H.

[0080] The electromagnet is controlled to conduct magnetic separation on the liquid H in each sample tube 6. After the liquid is discarded, the pipette 36 is controlled to inject different types of antibody diluent C into each sample tube 6 respectively, and the thermoelectric semiconductor refrigeration plate 25 in the reaction table 22 is controlled to cool (about 4°C). Then, the speed regulating motor is controlled to drive the sample tubes 6 in the reaction table 22 to flip overnight to complete the binding of the antibody and magnetic beads to obtain liquid I.

[0081] After magnetic separation of liquid I, discard the supernatant, control the pipette 36 to inject washing buffer B1 into each sample tube 6, and repeat this step three times. Then control the pipette 36 to inject different types of protein lysis buffer D into each sample tube 6, and control the reaction table 22 to flip. The user can select the flipping reaction time according to the substance binding situation, usually incubating at 4°C overnight to obtain liquid J.

[0082] Liquid J is magnetically separated, the supernatant is discarded, and the original sample tubes 6 are each washed with wash buffer B1. After five washes, the pipette 36 is controlled to inject protein loading buffer E1 into each sample tube 6 for resuspending, thereby obtaining a suspension. The thermoelectric semiconductor cooling element 25 within the reaction table 22 is then controlled to heat at 95°C for 10 minutes. The supernatant is then magnetically separated and collected for subsequent analysis.

[0083] The above is only an embodiment of the present invention, and the common knowledge such as the specific technical solutions and / or characteristics in the solution are not described in detail here. It should be pointed out that for those skilled in the art, without departing from the technical solution of the present invention, several variations and improvements can be made, which should also be regarded as the scope of protection of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the description can be used to interpret the content of the claims.

Claims

1. A multi-channel fully automatic intelligent protein interaction test device, characterized by: The invention comprises a test platform and a cover body arranged on the test platform. The test platform is provided with a protein reaction device, a pipetting device and a liquid storage device. The protein reaction device comprises a reaction table rotatably connected to the test platform. The reaction table is provided with a plurality of receiving holes for installing sample tubes. A magnetic piece for magnetically separating magnetic beads in the sample tubes is provided in the reaction table. The pipetting device is used for adding liquid to and aspirating liquid from the sample tubes and comprises a pipetting gun and a displacement mechanism for driving the pipetting gun to move horizontally and vertically. The liquid storage device is used for storing various experimental liquids at low temperatures and comprises a low-temperature liquid storage tank and a cover plate arranged on the low-temperature liquid storage tank. The low-temperature liquid storage tank is provided with a plurality of independent grooves for independently placing liquid boxes of different types of liquids.

2. A multi-channel fully automatic intelligent protein interaction test device according to claim 1, characterized in that: The reaction table is provided with a cooling module and a heating module corresponding to each receiving hole for cooling and heating the sample tubes. The top surface of the reaction table is rotatably connected to a flip plate, which is used to simultaneously open and close the tube openings of each sample tube. The flip plate is provided with a top heating module.

3. A multi-channel fully automatic intelligent protein interaction test device according to claim 2, characterized in that: The tube mouth of the sample tube is rotatably connected to a tube cover plate, which is provided with a cover protrusion for sealing the tube mouth and a cover opening hole. When the tube cover plate is in a state of sealing the tube mouth, the cover opening hole is misaligned with the end face of the tube mouth of the sample tube.

4. A multi-channel fully automatic intelligent protein interaction test device according to claim 3, characterized in that: The free end of the flip cover plate is provided with a cover opening mechanism corresponding to the cover opening hole, the cover opening mechanism comprising a connecting shaft and a cover opening member, the connecting shaft being vertically arranged on the lower surface of the flip cover plate, one end of the cover opening member being rotatably connected to the side wall of the connecting shaft, and the gravity of the cover opening member being greater than the rotational friction between the cover opening member and the connecting shaft; When the cover opening member is folded, the maximum transverse width of the cover opening mechanism is smaller than the diameter of the cover opening hole; when the cover opening member is opened, the maximum transverse width of the cover opening mechanism is larger than the diameter of the cover opening hole.

5. The multi-channel fully automatic intelligent protein interaction test device according to claim 4, characterized in that: The displacement mechanism includes a horizontal displacement unit and a vertical displacement unit. The horizontal displacement unit includes two parallel first slide rails and a second slide rail sliding on the two first slide rails. A fixed block is slidably connected to the second slide rail. The vertical displacement unit is connected to the fixed block. The pipette is provided on the vertical displacement unit. The vertical displacement unit is used to drive the pipette to move vertically.

6. The multi-channel fully automatic intelligent protein interaction test device according to claim 5, characterized in that: It also includes a gun tip processing mechanism, which includes a gun tip replacement unit and a gun tip recovery unit. The gun tip recovery unit is used to collect discarded gun tips of the pipette, and includes a waste channel and a collection box. The collection box is slidably connected to the side wall of the experimental platform, and the waste channel is tilted in the experimental platform and connected to the collection box; the gun tip replacement unit includes a gun tip box placement area and a camera. The camera is used to photograph the position of the gun tip in the gun tip box. The pipette moves to the designated position according to the gun tip position information and inserts the corresponding gun tip.

7. The multi-channel fully automatic intelligent protein interaction test device according to claim 6, characterized in that: The grooves are divided into multiple groove groups, wherein each groove in at least one groove group corresponds to placing various liquid boxes used in protein interaction experiments.

8. The multi-channel fully automatic intelligent protein interaction test device according to claim 7, characterized in that: The grooves in the groove group for placing magnetic bead suspensions are interconnected to form a magnetic bead liquid processing tank. A vortex oscillator is provided in the magnetic bead liquid processing tank. The vibration frame of the vortex oscillator is provided with multiple placement holes for placing magnetic bead liquid tubes in parallel. An opening cover plate is rotatably connected to the vibration frame. The opening cover plate is used to simultaneously open and close the tube openings of each magnetic bead liquid tube. The structure of the opening cover plate is the same as that of the flip cover plate, and the structure of the magnetic bead liquid tube is the same as that of the sample tube.

9. The multi-channel fully automatic intelligent protein interaction test device according to claim 8, characterized in that: It also includes a robotic arm, a marking instrument and a sample tube box placement area. The robotic arm is used to clamp the sample tube to a specified position, and the marking instrument is used to mark the sample tube.

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