WO2022166051A1 - 一种全自动wb孵育系统 - Google Patents

一种全自动wb孵育系统 Download PDF

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Publication number
WO2022166051A1
WO2022166051A1 PCT/CN2021/098551 CN2021098551W WO2022166051A1 WO 2022166051 A1 WO2022166051 A1 WO 2022166051A1 CN 2021098551 W CN2021098551 W CN 2021098551W WO 2022166051 A1 WO2022166051 A1 WO 2022166051A1
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fixed
fixing plate
plate
motor
incubation
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PCT/CN2021/098551
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English (en)
French (fr)
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周军
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南京宝沃生物科技有限公司
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Publication of WO2022166051A1 publication Critical patent/WO2022166051A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor

Definitions

  • the invention relates to a Western Blot (hereinafter referred to as WB) incubation instrument, in particular to an automatic WB incubation system.
  • WB Western Blot
  • WB technology namely Western blotting technology
  • WB technology is a basic and commonly used experimental method for analyzing biochemistry, molecular biology and immunogenetics, and is of great significance in the field of protein detection, analysis and research.
  • the experimental principle is that the primary antibody has a specific affinity reaction with the target protein attached to and immobilized on the chemically synthesized membrane support, and then binds with the enzyme or isotope-labeled secondary antibody, and finally develops the color by the substrate coloring. analysis, enabling qualitative and semi-quantitative measurements of proteins.
  • the present invention discloses a fully automatic WB incubation system, which realizes the fully automatic operation of WB incubation from membrane transfer to secondary antibody washing, optimizes the manual operation of traditional experimental process, and makes the experimental process more intelligent. Detect as many clinically meaningful antibodies as possible, reduce the time cost and labor cost of the experiment, eliminate the experimental error caused by human operation, and ensure the accuracy and repeatability of the experiment.
  • a fully automatic WB incubation system comprising an external control panel and a base plate; wherein a swing motor, a swing shaft support plate, a waste liquid tank and a reagent tube rack are arranged on the base plate in sequence; the rear side of the swing shaft support plate There is a column plate; the side of the column plate is fixed with a peristaltic pump fixing plate; wherein the first, second, third and fourth peristaltic pumps are all fixed on the peristaltic pump fixing plate; wherein the refrigeration component is fixed on the peristaltic pump Beside the plate, a beam is arranged on the column plate; a transverse movement mechanism is arranged on the beam.
  • the rear side of the vertical movement mechanism is connected and fixed with the lateral movement mechanism;
  • the beam is fixed on the column plate through the first reinforcing rib,
  • the swing motor fixing plate is fixedly installed on the bottom plate, and the first step motor and inductor Type proximity switches are respectively installed on the rocking motor fixing plate;
  • the output shaft of the first stepping motor is connected and fixed with the rocking link;
  • the rocking link is connected and fixed with the rocking shaft fixing block on the rocking fixed plate;
  • the rocking fixed A stainless steel pool is fixed on the plate and a rocking shaft fixing plate is fixed at the bottom;
  • the rocking shaft fixing plate is connected and fixed with the rocking shaft support plate fixed on the bottom plate through a rotating shaft; wherein the incubation pool is set in the stainless steel pool.
  • the reagent tube rack is sequentially provided with a spare pipette, a secondary antibody solution reagent tube and a primary antibody solution reagent tube;
  • the beam extension bracket on the vertical moving mechanism is connected and fixed with the fixing plate;
  • the fixing plate is provided with a slider track; the rear end of the slider is slidably connected with the slider track and the front end is connected and fixed with the motor nut seat;
  • the screw motor is fixed on the top of the fixing plate and is connected and fixed with the screw on the fixing plate;
  • the motor nut seat is connected with the screw
  • the rod is connected and fixed, and the upper and lower ends of one side are respectively provided with a first limit piece and a second limit piece;
  • the fixed plate is provided with a first photosensitive switch and a second photoelectric induction switch; wherein the pipette shaft It is fixed on the underside of the motor nut seat, the suction head is connected with the suction pipe shaft, and the two stainless steel needles are fixed on the bottom of the fixing plate.
  • the lateral movement mechanism includes a second stepping motor and a guide rail; wherein the second stepping motor is fixed on the beam and the output shaft is provided with a first synchronizing wheel; wherein the first synchronizing wheel is synchronized by The belt is connected and fixed with the second synchronizing wheel on the upper end of the beam; the guide rail is fixed on the beam, the rear side of the belt fixing plate is connected and fixed with the timing belt, and the front end is connected and fixed with the beam extension bracket; the lower side of the beam Both ends are respectively fixed with a third photoelectric induction switch and a fourth photoelectric induction switch; one end of the beam extension bracket is provided with a third limiting piece.
  • a further improvement of the present invention is that: the side surface of the stainless steel pool is provided with a water level sensor and a temperature sensor, and an ultrasonic vibrator is fixed at the lower part.
  • the invention is further improved in that the first peristaltic pump is connected to the pipette shaft, the second peristaltic pump is connected to the stainless steel pool, and the third and fourth peristaltic pumps are respectively connected to two stainless steel needles.
  • the further improvement of the present invention lies in that: the reagent tube rack is fixed on the bottom plate through equal height columns.
  • a further improvement of the present invention is that the refrigeration assembly is located beside the peristaltic pump fixing plate and is electrically connected with the operation control panel.
  • the refrigeration component is located beside the fixed plate of the peristaltic pump; the data information from the temperature sensor is collected by operating the components inside the control panel, and the data is processed, and then the internal program is triggered to control the refrigeration component to work, thereby
  • the temperature in the fully automatic WB incubation system is maintained at the temperature required for incubation, and the refrigeration component forms a closed-loop control with the entire system to achieve constant temperature control.
  • the first step motor drives the rotation of the motor turntable, and then drives the rocking shaft fixing block connected with the rocking link to move, and finally makes the rocking fixed plate connected with the rocking shaft fixing block to perform a certain angle of rocking motion, so as to achieve the The effect of rocking the incubation pool on the immobilization plate accelerates the incubation.
  • the stainless steel pool fixed on the rocking fixed plate provides a water bath environment for the incubation pool.
  • the temperature sensor installed on the side of the stainless steel pool can ensure real-time monitoring of the temperature of the incubation pool, and the water level sensor can ensure that the water in the stainless steel pool will not overflow. .
  • An ultrasonic vibrator is fixed at the lower part of the stainless steel pool.
  • the ultrasonic vibrator efficiently converts the electrical energy into the high-frequency vibration of its own mechanical components, which is transmitted to the water itself in the stainless steel pool, which drives the high-frequency vibration of the water, and then transmits the vibration to the medium. , to speed up the binding of antibody molecules in the primary antibody or secondary antibody solution to the specific target protein, so as to speed up the incubation of the membrane sheets in the incubation pool.
  • the WBA automatic WB incubation system controls the second stepper motor to drive the first synchronous wheel to rotate through the external operation panel, so as to drive the movement of the synchronous belt, and at the same time to synchronize with
  • the belt fixing plate connected with the belt moves laterally, thereby driving the beam extension bracket fixed with it to move, and then driving the screw motor part to move laterally, and the third limit piece reaches the third photoelectric induction switch position, which is the entire
  • the leftmost end that can be reached by the beam extension bracket, when the limit piece reaches the position of the fourth photoelectric sensor switch, this position is the rightmost position that the beam extension bracket can reach, corresponding to the entire screw motor part. left and right limit position;
  • the control panel to control the rotation of the screw motor, so that the motor nut seat drives the slider to move along the slider track, and at the same time drives the straw shaft to move longitudinally; in the working state, the second stepper motor controls the screw motor part
  • the screw motor controls the longitudinal movement of the suction head and the stainless steel needle; after working for a period of time, the second stepper motor and the screw motor work together to move the suction head to the upper part of the incubation pool, and the stepper motor is also inductive Under the action of the proximity switch, the incubation tank is tilted, so that the suction head can remove all the blocking liquid under the action of the first peristaltic pump, and then all the blocking liquid is injected into the waste liquid tank.
  • the fourth peristaltic pump works, injects the cleaning solution into the incubation tank through another stainless steel needle to clean the residual blocking solution, and then sucks the liquid in the incubation tank through the suction head and injects it into the waste liquid tank.
  • the screw motor The part reaches the right limit position under the action of the second stepper motor. The screw motor starts to work.
  • the addition of the primary antibody solution and the secondary antibody solution is carried out by moving the tip to the position on the corresponding reagent tube for suction and then adding it to the incubation pool. After that, the solution is removed, and the cleaning solution is added and removed. The whole process of detachment recovery and re-clamping and addition and removal of blocking solution is the same.
  • the present invention has the beneficial effects of realizing fully automatic operation of WB incubation from membrane transfer to secondary antibody washing, optimizing the manual operation of traditional experimental process, making the experimental process more intelligent, and improving work efficiency.
  • FIG. 1 the structural representation of the present invention
  • FIG. 1 the structural schematic diagram of the lower end in Figure 1;
  • FIG. 3 the structural schematic diagram of the lateral movement mechanism
  • Fig. 4 the structural schematic diagram of the vertical movement mechanism
  • 1- second stepper motor 2- first synchronous wheel, 3- synchronous belt, 4- beam, 5- third photoelectric sensor switch, 6- first peristaltic pump, 7- second peristaltic pump, 8- Peristaltic pump fixing plate, 9-third peristaltic pump, 10-fourth peristaltic pump, 11-refrigeration component, 12-second reinforcing rib, 13-bottom plate, 14-incubation pool, 15-stainless steel pool, 16-swing fixing plate , 17- rocking shaft support plate, 18- rocking axis fixing plate, 19- ultrasonic vibrator, 20- inductive proximity switch, 21- motor turntable, 22- rocking motor fixing plate, 23- rocking connecting rod, 24- first step Inlet motor, 25-swing shaft fixing block, 26-water level sensor, 27-temperature sensor, 28-waste tank, 29-equal height column, 30-primary antibody solution reagent tube, 31-reagent tube rack, 32-secondary antibody Solution reagent tube, 33-spare tip, 34-tip,
  • a fully automatic WB incubation system of this embodiment includes an operation control panel 55 and a bottom plate 13; wherein the bottom plate 13 is provided with a swing motor fixing plate 22, a swing shaft support plate 17, The waste liquid tank 28 and the reagent tube rack 31; the rear side of the rocking shaft support plate 17 is provided with a column plate 36; the side of the column plate 36 is fixed with a peristaltic pump fixing plate 8 and is fixed by the second reinforcing rib 12 On the bottom plate 13; wherein the first, second, third and fourth peristaltic pumps 6, 7, 9, 10 are all fixed on the peristaltic pump fixing plate 8; wherein the refrigeration assembly 11 is located next to the peristaltic pump fixing plate 8,
  • the column plate 36 is provided with a beam 4; the beam 4 is provided with a lateral movement mechanism; the rear side of the vertical movement mechanism is connected and fixed with the lateral movement mechanism; On the column plate 36 , the reagent tube rack 31 is fixed on the bottom plate 13 through the equal height column 29 .
  • the swing motor fixing plate 22 is fixedly installed on the bottom plate 13, the first stepping motor 24 and the inductive proximity switch 20 are respectively installed on the swing motor fixing plate 22; the output shaft of the first stepping motor 24 and the swing connecting rod 23 connection and fixation; the swing link 23 is connected and fixed with the swing shaft fixing block 25 on the swing fixing plate 16; the swing fixing plate 16 is fixed with a stainless steel pool 15 and the bottom is fixed with a swing shaft fixing plate 18; the The rocking shaft fixing plate 18 is connected and fixed with the rocking shaft supporting plate 17 fixed on the bottom plate 13 through the rotating shaft; wherein the incubation pool 14 is arranged inside the stainless steel pool 15; the side of the stainless steel pool 15 is provided with a water level sensor 26 The ultrasonic vibrator 19 is fixed to the lower part of the temperature sensor 27 .
  • the reagent tube rack 31 is provided with a spare pipette 33, a secondary antibody solution reagent tube 32 and a primary antibody solution reagent tube 30 in sequence;
  • the refrigeration assembly 11 is located next to the peristaltic pump fixing plate 8; the data information from the temperature sensor 27 is collected by operating the components inside the control panel 55, and the data is processed, and then the internal program is triggered to control the refrigeration assembly 11 to perform Work, so that the temperature in the fully automatic WB incubation system is maintained at the temperature required for incubation, and the refrigeration component forms a closed-loop control with the entire system to achieve constant temperature control.
  • the refrigeration assembly of the present invention that is, the air-to-air refrigeration assembly, refers to that the heat transfer mode of the cold end and the hot end is the air mode, that is, the heat dissipation mode of the radiator (or heat pipe) and the fan is adopted.
  • Refrigeration components are usually used to cool a closed cavity, or to help transfer heat generated by objects in the cavity to the outside of the cavity. This achieves the purpose of cooling the entire system.
  • the beam extension bracket 50 on the vertical moving mechanism is connected and fixed with the fixing plate 49 ;
  • the fixing plate 49 is provided with a slider track 48 ;
  • the rear end of the slider 43 is connected to the slider track 48 Sliding connection and the front end is connected and fixed with the motor nut seat 41;
  • the screw motor 42 is fixed on the top of the fixing plate 49 and is connected and fixed with the screw on the fixing plate 49;
  • the motor nut seat 41 is connected and fixed with the screw and one of the upper , the lower ends are respectively provided with a first limit piece 46 and a second limit piece 47;
  • the fixing plate 49 is provided with a first photosensitive switch 44 and a second photoelectric induction switch 45; wherein the pipette shaft 40 is fixed on the motor On the lower side of the nut seat 41, the suction head 34 is connected with the suction pipe shaft 40, and the two stainless steel needle heads 35 are fixed on the bottom of the fixing plate 49;
  • the lateral movement mechanism includes a second stepping motor 1 and a guide rail 38 ; wherein the second stepping motor 1 is fixed on the beam 4 through a fixing plate, and the output shaft is provided with a first synchronizing wheel 2 Wherein the first synchronizing wheel 2 is connected and fixed with the second synchronizing wheel 39 on the upper end of the beam 4 through the synchronous belt 3; wherein the guide rail 38 is fixed on the beam 4, and the rear side of the belt fixing plate 52 is connected with the synchronous belt 3 Fixed and the front end is connected and fixed with the beam extension bracket 50, and the rear side of the beam extension bracket 50 is connected with the slider 53; the lower ends of the beam 4 are respectively fixed with a third photoelectric induction switch 5 and a fourth photoelectric induction switch 37 ; One end of the beam extension bracket 50 is provided with a third limiting piece 54 .
  • the second stepper motor 1 works to drive the incubation pool 14 to swing.
  • the first step motor 24 drives the rotary motion of the motor turntable 21, and then drives the rocking shaft fixing block 25 connected with the rocking link 23 to move, and finally makes the rocking fixed plate 16 connected with the rocking shaft fixing block 25 to perform a certain angle of rocking motion. , so as to achieve the effect of accelerating the incubation of the incubation pool 14 on the rocking fixed plate 16 .
  • the stainless steel pool 15 fixed on the swing fixing plate 16 provides a water bath environment for the incubation pool 14.
  • the temperature sensor 27 installed on the side of the stainless steel pool 15 can ensure real-time monitoring of the temperature of the incubation pool, and the water level sensor 26 can ensure the stainless steel pool. The water in 15 will not overflow.
  • An ultrasonic vibrator 19 is fixed at the lower part of the stainless steel pool 15, and the ultrasonic vibrator 19 efficiently converts the electrical energy into the high-frequency vibration of its own mechanical components, thereby conducting the water itself in the stainless steel pool 15, driving the high-frequency vibration of the water, and then the The vibration is transmitted to 14 to accelerate the binding of antibody molecules in the primary antibody or secondary antibody solution to the specific target protein, so as to speed up the incubation of the membrane in the incubation pool.
  • the WBA automatic WB incubation system controls the second stepper motor 1 through the external operation panel to drive the first synchronizing wheel 2 to rotate, so as to Drive the movement of the synchronous belt 3, and at the same time make the belt fixing plate 52 connected with the synchronous belt 3 move laterally, thereby driving the beam extension bracket 50 fixed with it to move, and then driving the screw motor 42 to move laterally.
  • the limit piece 54 reaches the position of the third photoelectric induction switch 5, which is the leftmost end that the entire beam extension bracket 50 can reach.
  • the third limit piece 54 reaches the position of the fourth photoelectric induction switch 37, this position is the extension of the beam
  • the rightmost position that the bracket 50 can reach corresponds to the left and right extreme positions that the entire screw motor part can reach.
  • the control panel is used to control the rotation of the screw motor 42, so that the motor nut seat 41 drives the slider 43 to move along the slider track 48, and at the same time drives the straw shaft 40 to move longitudinally; in the working state
  • the second stepping motor 1 controls the lateral movement of the screw motor part
  • the screw motor 42 controls the longitudinal movement of the suction head 34 and the stainless steel needle 35; after working for a period of time, the second stepping motor 1 and the screw motor 42 work together
  • the first step motor 24 is also under the action of the inductive proximity switch 20, so that the incubation pool 14 is tilted, so that the suction head can be easily moved under the action of the first peristaltic pump 6. All the blocking liquid is removed, and then all the blocking liquid is poured into the waste liquid tank 28 .
  • the fourth peristaltic pump 10 works, injects the cleaning solution into the incubation tank 14 through another stainless steel needle 35 to clean the remaining sealing solution, and then sucks the liquid in the incubation tank through the suction head 34 and injects it into the waste liquid tank 28 , and finally, the screw motor part reaches the right limit position under the action of the second stepper motor 1 .
  • the screw motor 42 starts to work.
  • the suction head 34 and the suction pipe shaft 40 start In preparation for disengagement, when the first limiting piece 46 reaches the position of the first photoelectric sensor switch 44, the suction head 34 and the pipette shaft 40 are officially disengaged, and the used suction head will be separated and dropped into a waste test tube slot. At this point, the adding and cleaning process of the blocking solution is all over.
  • the suction pipe shaft 40 moves to the position of the spare suction head 33, and the screw motor 42 works, so that the suction pipe shaft 40 falls to a certain position, so that the spare suction head 33 is fixed on the suction pipe shaft 40, and is prepared for the addition of the subsequent solution.
  • the addition of the primary antibody solution and the secondary antibody solution is carried out by moving the tip 34 to the position on the corresponding reagent tube for suction and then adding it to the incubation pool. After that, the solution is removed, the cleaning solution is added and removed, and the tip is detached. Recovery and re-clamping and the addition and removal of blocking solution are the same as the whole process.

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Abstract

本发明提供一种全自动WB孵育系统，包括外置的控制面板和底板；其中所述底板上依次设有摇摆电机、摇摆轴支撑板、废液槽和试剂管架；所述摇摆轴支撑板的后侧设有立柱板；所述立柱板的侧边固定设有蠕动泵固定板；所述立柱板上设有横梁；所述横梁上设有横向移动机构，其中竖向移动机构的后侧与所述横向移动机构连接固定；其中孵育池设置在所述不锈钢水池的内部；其中试剂管架上依次设有备用吸头、二抗溶液试剂管和一抗溶液试剂管；竖向移动机构上的横梁延伸支架与固定板连接固定，本发明实现WB孵育从转膜后续到二抗洗涤的全自动化操作，减少实验的时间成本和人工成本，排除人为操作带入的实验误差，保证实验的准确性和可重复性。

Description

一种全自动WB孵育系统 技术领域
本发明涉及一种Western Blot(以下简称WB)孵育仪器，具体是一种全自动WB孵育系统。
背景技术
WB技术即蛋白印迹技术是分析生物化学、分子生物学和免疫遗传学的基础的、常用的实验方法，在蛋白质检测、分析和研究领域具有重要意义。其实验原理是通过第一抗体与附着、固定在化学合成膜支持物上的靶蛋白发生特异性亲和反应，再与酶或同位素标记的第二抗体结合，最后通过底物显色进行显色分析，从而实现对蛋白质定性和半定量的测量。
传统的Western Blot实验需进行制胶、电泳、转膜、封闭、一抗、洗涤、二抗、洗涤、发光等繁琐步骤。从封闭步骤开始，实验人员将转印后的膜片放进装有5％奶粉封闭液的密封盒中，用以封闭非特异性结合位点。1-2h后倒掉封闭液，用TBST清洗1min，倒掉TBST后，加入一抗溶液，在4℃的环境下过夜孵育或室温下孵育2h，用以第一抗体与特异性目的蛋白结合。一抗孵育结束后TBST清洗一抗溶液10min，3次。再加入二抗溶液，室温环境下孵育1h，结束后TBST清洗10min，3次。最后进行底物显色曝光等处理。传统的实验方法，是由人为进行时间的估量与判断，费事费力，极大影响工作效率，并且频繁对膜片进行手工处理会造成实验的污染与误差，鉴于以上的缺 陷，实有必要设计一款WBA全自动WB孵育系统。
发明内容
为解决上述问题，本发明公开了一种全自动WB孵育系统，实现WB孵育从转膜后续到二抗洗涤的全自动化操作，优化传统实验过程人工操作这些步骤，让实验过程更加的智能化，尽可能多的检测出具有临床意义的抗体，减少实验的时间成本和人工成本，排除人为操作带入的实验误差，保证实验的准确性和可重复性。
一种全自动WB孵育系统，包括外置的控制面板和底板；其中所述底板上依次设有摇摆电机、摇摆轴支撑板、废液槽和试剂管架；所述摇摆轴支撑板的后侧设有立柱板；所述立柱板的侧边固定设有蠕动泵固定板；其中第一、二、三、四蠕动泵均固定在所述蠕动泵固定板；其中制冷组件位于所述蠕动泵固定板旁，所述立柱板上设有横梁；所述横梁上设有横向移动机构。
其中竖向移动机构的后侧与所述横向移动机构连接固定；其中横梁通过第一加强筋固定在所述立柱板上，其中摇摆电机固定板固定安装在底板上，第一步进电机和电感式接近开关分别安装在摇摆电机固定板上；其中第一步进电机的输出轴与摇摆连杆连接固定；所述摇摆连杆与摇摆固定板上的摇摆转轴固定块连接固定；所述摇摆固定板上固定设有不锈钢水池且底部固定设有摇摆轴固定板；所述摇摆轴固定板通过转轴与固定在所述底板上的摇摆轴支撑板连接固定；其中孵育池设置在所述不锈钢水池的内部；其中试剂管架上依次设有备用吸 头、二抗溶液试剂管和一抗溶液试剂管；竖向移动机构上的横梁延伸支架与固定板连接固定；所述固定板上设有滑块轨道；其中滑块的后端与所述滑块轨道滑动连接且前端与电机螺母座连接固定；丝杆电机固定在固定板的顶部且与固定板上的丝杆连接固定；电机螺母座与丝杆连接固定且其中一侧的上、下两端分别设有第一限位片和第二限位片；所述固定板上设有第一光感应开关和第二光电感应开关；其中吸管轴固定在电机螺母座的下侧，吸头与吸管轴连接，两个不锈钢针头均固定在固定板的底部。
本发明进一步改进在于：所述横向移动机构包括第二步进电机和导轨；其中第二步进电机固定在所述横梁上且输出轴上设有第一同步轮；其中第一同步轮通过同步带与所述横梁上端部的第二同步轮连接固定；其中导轨固定在所述横梁上，皮带固定板的后侧与同步带连接固定且前端与横梁延伸支架连接固定；所述横梁的下侧两端分别固定设有第三光电感应开关和第四光电感应开关；所述横梁延伸支架的一端设有第三限位片。
本发明进一步改进在于：所述不锈钢水池的侧面设有水位传感器和温度传感器且下部固定有超声波振子。
本发明进一步改进在于：其中第一蠕动泵连接吸管轴，第二蠕动泵连接不锈钢水池，第三、四蠕动泵分别连接两个不锈钢针头。
本发明进一步改进在于：试剂管架通过等高柱固定在所述底板上。
本发明进一步改进在于：其中制冷组件位于所述蠕动泵固定板旁 并与所述操作控制面板电性连接。
本发明工作时：
1、其中制冷组件位于所述蠕动泵固定板旁；通过操作控制面板内部的元器件采集来自温度传感器的数据信息，并进行数据的处理，进而触发内部的程序，以控制制冷组件进行工作，从而使得全自动WB孵育系统中的温度保持在孵化所需要的温度下，制冷组件与整个系统形成闭环控制，实现温度的恒定控制。
2、在工作初始，人工的将转膜后的膜片放入孵育池中，将吸头运动到孵育池上部，蠕动泵工作，通过不锈钢针头向孵育池中注入封闭液，同时第二步进电机工作，带动孵育池进行摇摆。
3、第一步进电机带动电机转盘的旋转运动，进而带动与摇摆连杆连接的摇摆转轴固定块运动，最终使得与摇摆转轴固定块连接的摇摆固定板进行一定角度的摇摆运动，以达到让摇摆固定板上的孵育池进行加速孵育的效果。其中摇摆固定板上固定的不锈钢水池为孵育池提供水浴的环境，在不锈钢水池的侧面装有的温度传感器，可以保证对孵育池的温度实时监控，水位传感器可以保证不锈钢水池中的水不会溢出。在不锈钢水池下部固定有超声波振子，超声波振子将电能高效率地转化为其本身机械部件的高频率振动，从而传导到不锈钢水池中的水本身，带动水的高频振动，进而将振动传导到中，加快一抗或者二抗溶液中抗体分子与特异性目的蛋白结合，以加快孵育池中膜片孵育的速度。
4、由于设有横向移动机构，能够实现左右横向移动，WBA全自 动WB孵育系统通过外接的操作面板控制第二步进电机带动第一同步轮旋转，以带动同步带的移动，同时使得与同步带相连接的皮带固定板横向移动，从而带动和其连固定的横梁延伸支架移动，进而带动丝杆电机部分进行横向移动，在第三限位片到达第三光电感应开关位置，这个位置是整个横梁延伸支架所能到达的最左端，当限位片到达第四光电感应开关的位置，这个位置是横梁延伸支架所能到达的最右端位置，相对应的也是整个丝杆电机部分所能到达的左右极限位置；
利用控制面板控制丝杆电机的旋转，使得电机螺母座带动滑块沿着滑块轨道进行移动，并同时带动吸管轴进行纵向的移动；在工作状态下，第二步进电机控制丝杆电机部分的横向移动，丝杆电机控制着吸头和不锈钢针头纵向移动；工作一段时间后，第二步进电机和丝杆电机共同作用，将吸头移动到孵育池上部，步进电机也在电感式接近开关的作用下，使得孵育池进行倾斜，以方便吸头能够在第一蠕动泵的作用下，将封闭液全部清除，然后将封闭液全部注入废液槽中。
然后第四蠕动泵工作，通过另外一根不锈钢针头向孵育池中注入清洗液清洗残余的封闭液，再通过吸头将孵育池中的液体吸出，注入到废液槽中，最后，丝杆电机部分在第二步进电机的作用下到达右极限位置。丝杆电机开始工作，在第二限位片到达第二光电感应开关，而第一限位片未到达光第一电感应开关位置的时候，吸头和吸管轴开始进入脱离准备，当第一限位片到达第一光电感应开关的位置，吸头和吸管轴正式脱离，使用过的吸头将会被分离脱落到一个废弃试管槽内。至此，封闭液的添加与清洗过程全部结束。吸管轴运动到备用吸 头的位置，丝杆电机工作，使吸管轴下落到一定位置，使得备用吸头固定到吸管轴上，为后续溶液的添加做工作准备。
5、对应的，一抗溶液，二抗溶液的添加是由吸头运动到对应试剂管上的位置进行吸取然后加入到孵育池中的，之后溶液的清除，清洗液的添加和去除，吸头的脱离回收和再次装夹和封闭液的添加去除整个过程一样。
本发明的有益效果：实现WB孵育从转膜后续到二抗洗涤的全自动化操作，优化传统实验过程人工操作这些步骤，让实验过程更加的智能化，提高工作效率。
附图说明
图1、本发明的结构示意图；
图2、图1中下端部的结构示意图；
图3、横向移动机构的结构示意图；
图4、竖向移动机构的结构示意图；
图5、各蠕动泵连接示意图；
附图标记列表：
其中：1-第二步进电机、2-第一同步轮、3-同步带、4-横梁、5-第三光电感应开关、6-第一蠕动泵、7-第二蠕动泵、8-蠕动泵固定板、9-第三蠕动泵、10-第四蠕动泵、11-制冷组件、12-第二加强筋、13-底板、14-孵育池、15-不锈钢水池、16-摇摆固定板、17-摇摆轴支撑板、18-摇摆轴固定板、19-超声波振子、20-电感式接近开关、21- 电机转盘、22-摇摆电机固定板、23-摇摆连杆、24-第一步进电机、25-摇摆转轴固定块、26-水位传感器、27-温度传感器、28-废液槽、29-等高柱、30-一抗溶液试剂管、31-试剂管架、32-二抗溶液试剂管、33-备用吸头、34-吸头、35-不锈钢针头、36-立柱板、37-第四光电感应开关、38-导轨、39-第二同步轮、40-吸管轴、41-电机螺母座、42-丝杆电机、43-滑块、44-第一光电感应开关、45-第二光电感应开关、46-第一限位片、47-第二限位片、48-滑块轨道、49-固定板、50-横梁延伸支架、51-第一加强筋、52-皮带固定板、53-滑块、54-第三限位片、55-操作控制面板。
具体实施方式
下面结合附图和具体实施方式，进一步阐明本发明，应理解下述具体实施方式仅用于说明本发明而不用于限制本发明的范围。需要说明的是，下面描述中使用的词语“前”、“后”、“左”、“右”、“上”和“下”指的是附图中的方向，词语“内”和“外”分别指的是朝向或远离特定部件几何中心的方向。
如图1-5所示，本实施例的一种全自动WB孵育系统，包括操作控制面板55和底板13；其中所述底板13上依次设有摇摆电机固定板22、摇摆轴支撑板17、废液槽28和试剂管架31；所述摇摆轴支撑板17的后侧设有立柱板36；所述立柱板36的侧边固定设有蠕动泵固定板8且通过第二加强筋12固定在所述底板13上；其中第一、二、三、四蠕动泵6、7、9、10均固定在所述蠕动泵固定板8；其中 制冷组件11位于所述蠕动泵固定板8旁，所述立柱板36上设有横梁4；所述横梁4上设有横向移动机构；其中竖向移动机构的后侧与所述横向移动机构连接固定；其中横梁4通过第一加强筋51固定在所述立柱板36上，其中试剂管架31通过等高柱29固定在所述底板13上。
其中摇摆电机固定板22固定安装在底板13上，第一步进电机24和电感式接近开关20分别安装在摇摆电机固定板22上；其中第一步进电机24的输出轴与摇摆连杆23连接固定；所述摇摆连杆23与摇摆固定板16上的摇摆转轴固定块25连接固定；所述摇摆固定板16上固定设有不锈钢水池15且底部固定设有摇摆轴固定板18；所述摇摆轴固定板18通过转轴与固定在所述底板13上的摇摆轴支撑板17连接固定；其中孵育池14设置在所述不锈钢水池15的内部；所述不锈钢水池15的侧面设有水位传感器26和温度传感器27且下部固定有超声波振子19。
其中试剂管架31上依次设有备用吸头33、二抗溶液试剂管32和一抗溶液试剂管30；
其中制冷组件11位于所述蠕动泵固定板8旁；通过操作控制面板55内部的元器件采集来自温度传感器27的数据信息，并进行数据的处理，进而触发内部的程序，以控制制冷组件11进行工作，从而使得全自动WB孵育系统中的温度保持在孵化所需要的温度下，制冷组件与整个系统形成闭环控制，实现温度的恒定控制。本发明的制冷组件，即空气对空气型制冷组件，指冷端和热端的传热方式均为空气的 方式，即均采用散热器(或热管)和风扇的散热方式。制冷组件通常用来给密闭的腔体降温，或者是帮助把腔体内物体产生的热量转移到腔体外。由此达到给整个系统降温的目的。
如图4所示：竖向移动机构上的横梁延伸支架50与固定板49连接固定；所述固定板49上设有滑块轨道48；其中滑块43的后端与所述滑块轨道48滑动连接且前端与电机螺母座41连接固定；丝杆电机42固定在固定板49的顶部且与固定板49上的丝杆连接固定；电机螺母座41与丝杆连接固定且其中一侧的上、下两端分别设有第一限位片46和第二限位片47；所述固定板49上设有第一光感应开关44和第二光电感应开关45；其中吸管轴40固定在电机螺母座41的下侧，吸头34与吸管轴40连接，两个不锈钢针头35均固定在固定板49的底部；
如图5所示；其中第一蠕动泵6蠕动泵连接吸管轴40，第二蠕动泵7连接不锈钢水池15，第三、四蠕动泵9、10分别连接两个不锈钢针头35。
如图1所示：所述横向移动机构包括第二步进电机1和导轨38；其中第二步进电机1通过固定板固定在所述横梁4上且输出轴上设有第一同步轮2；其中第一同步轮2通过同步带3与所述横梁4上端部的第二同步轮39连接固定；其中导轨38固定在所述横梁4上，皮带固定板52的后侧与同步带3连接固定且前端与横梁延伸支架50连接固定，而横梁延伸支架50后侧与滑块53连接；所述横梁4的下侧两端分别固定设有第三光电感应开关5和第四光电感应开关37；所述 横梁延伸支架50的一端设有第三限位片54。
在工作初始，人工的将转膜后的膜片放入孵育池中，将吸头34运动到孵育池14上部；对应的蠕动泵工作，通过不锈钢针头35向孵育池14中注入封闭液，同时第二步进电机1工作，带动孵育池14进行摇摆。第一步进电机24带动电机转盘21的旋转运动，进而带动与摇摆连杆23连接的摇摆转轴固定块25运动，最终使得与摇摆转轴固定块25连接的摇摆固定板16进行一定角度的摇摆运动，以达到让摇摆固定板16上的孵育池14进行加速孵育的效果。其中摇摆固定板16上固定的不锈钢水池15为孵育池14提供水浴的环境，在不锈钢水池15的侧面装有的温度传感器27，可以保证对孵育池的温度实时监控，水位传感器26可以保证不锈钢水池15中的水不会溢出。在不锈钢水池15下部固定有超声波振子19，超声波振子19将电能高效率地转化为其本身机械部件的高频率振动，从而传导到不锈钢水池15中的水本身，带动水的高频振动，进而将振动传导到14中，加快一抗或者二抗溶液中抗体分子与特异性目的蛋白结合，以加快孵育池中膜片孵育的速度。
由于设有横向移动机构和竖向移动机构，能够实现左、右横向移动和上下移动；WBA全自动WB孵育系统通过外接的操作面板控制第二步进电机1带动第一同步轮2旋转，以带动同步带3的移动，同时使得与同步带3相连接的皮带固定板52横向移动，从而带动和其连固定的横梁延伸支架50移动，进而带动丝杆电机42部分进行横向移动，在第三限位片54到达第三光电感应开关5位置，这个位置是整 个横梁延伸支架50所能到达的最左端，当第三限位片54到第四光电感应开关37的位置，这个位置是横梁延伸支架50所能到达的最右端位置，相对应的也是整个丝杆电机部分所能到达的左右极限位置。
在竖向移动机构中，利用控制面板控制丝杆电机42的旋转，使得电机螺母座41带动滑块43沿着滑块轨道48进行移动，并同时带动吸管轴40进行纵向的移动；在工作状态下，第二步进电机1控制丝杆电机部分的横向移动，丝杆电机42控制着吸头34和不锈钢针头35纵向移动；工作一段时间后，第二步进电机1和丝杆电机42共同作用，将吸头移动到孵育池上部，第一步进电机24也在电感式接近开关20的作用下，使得孵育池14进行倾斜，以方便吸头能够在第一蠕动泵6的作用下，将封闭液全部清除，然后将封闭液全部注入废液槽28中。
而后，第四蠕动泵10工作，通过另外一根不锈钢针头35向孵育池14中注入清洗液清洗残余的封闭液，再通过吸头34将孵育池中的液体吸出，注入到废液槽28中，最后，丝杆电机部分在第二步进电机1的作用下到达右极限位置。丝杆电机42开始工作，在第二限位片47到达第二光电感应开关45，而第一限位片46未到达光第一电感应开关44位置的时候，吸头34和吸管轴40开始进入脱离准备，当第一限位片46到达第一光电感应开关44的位置，吸头34和吸管轴40正式脱离，使用过的吸头将会被分离脱落到一个废弃试管槽内。至此，封闭液的添加与清洗过程全部结束。吸管轴40运动到备用吸头33的位置，丝杆电机42工作，使吸管轴40下落到一定位置，使 得备用吸头33固定到吸管轴40上，为后续溶液的添加做工作准备。
对应的，一抗溶液，二抗溶液的添加是由吸头34运动到对应试剂管上的位置进行吸取然后加入到孵育池中，之后溶液的清除，清洗液的添加和去除，吸头的脱离回收和再次装夹和封闭液的添加去除整个过程一样。
本发明方案所公开的技术手段不仅限于上述实施方式所公开的技术手段，还包括由以上技术特征任意组合所组成的技术方案。

Claims (6)

  1. 一种全自动WB孵育系统，其特征在于：包括操作控制面板(55)和底板(13)；其中所述底板(13)上依次设有摇摆电机固定板(22)、摇摆轴支撑板(17)、废液槽(28)和试剂管架(31)；所述摇摆轴支撑板(17)的后侧设有立柱板(36)；所述立柱板(36)的侧边固定设有蠕动泵固定板(8)；其中第一、二、三、四蠕动泵(6、7、9、10)均固定在所述蠕动泵固定板(8)；所述立柱板(36)上设有横梁(4)；所述横梁(4)上设有横向移动机构；其中竖向移动机构的后侧与所述横向移动机构连接固定；其中横梁(4)通过第一加强筋(51)固定在所述立柱板(36)上，其中摇摆电机固定板(22)固定安装在底板(13)上，第一步进电机(24)和电感式接近开关(20)分别安装在摇摆电机固定板(22)上；其中第一步进电机(24)的输出轴与摇摆连杆(23)连接固定；所述摇摆连杆(23)与摇摆固定板(16)上的摇摆转轴固定块(25)连接固定；所述摇摆固定板(16)上固定设有不锈钢水池(15)且底部固定设有摇摆轴固定板(18)；所述摇摆轴固定板(18)通过转轴与固定在所述底板(13)上的摇摆轴支撑板(17)连接固定；其中孵育池(14)设置在所述不锈钢水池(15)的内部；其中试剂管架(31)上依次设有备用吸头(33)、二抗溶液试剂管(32)和一抗溶液试剂管(30)；竖向移动机构上的横梁延伸支架(50)与固定板(49)连接固定；所述固定板(49)上设有滑块轨道(48)；其中滑块(43)的后端与所述滑块轨道(48)滑动连接且前端与电机螺母座(41)连接固定；丝杆电机(42)固定在 固定板(49)的顶部且与固定板(49)上的丝杆连接固定；电机螺母座(41)与丝杆连接固定且其中一侧的上、下两端分别设有第一限位片(46)和第二限位片(47)；所述固定板(49)上设有第一光感应开关(44)和第二光电感应开关(45)；其中吸管轴(40)固定在电机螺母座(41)的下侧，吸头(34)与吸管轴(40)连接，两个不锈钢针头(35)均固定在固定板(49)的底部。
  2. 根据权利要求1所述的一种全自动WB孵育系统，其特征在于：所述横向移动机构包括第二步进电机(1)和导轨(38)；其中第二步进电机(1)固定在所述横梁(4)上且输出轴上设有第一同步轮(2)；其中第一同步轮(2)通过同步带(3)与所述横梁(4)上端部的第二同步轮(39)连接固定；其中导轨(38)固定在所述横梁(4)上，皮带固定板(52)的后侧与同步带(3)连接固定且前端与横梁延伸支架(50)连接固定；所述横梁(4)的下侧两端分别固定设有第三光电感应开关(5)和第四光电感应开关(37)；所述横梁延伸支架(50)的一端设有第三限位片(54)。
  3. 根据权利要求1所述的一种全自动WB孵育系统，其特征在于：所述不锈钢水池(15)的侧面设有水位传感器(26)和温度传感器(27)且下部固定有超声波振子(19)。
  4. 根据权利要求1所述的一种全自动WB孵育系统，其特征在于：其中第一蠕动泵(6)连接吸管轴(40)，第二蠕动泵(7)连接不锈钢水池(15)，第三、四蠕动泵(9、10)分别连接两个不锈钢针头(35)。
  5. 根据权利要求1所述的一种全自动WB孵育系统，其特征在于：试 剂管架(31)通过等高柱(29)固定在所述底板(13)上。
  6. 根据权利要求1所述的一种全自动WB孵育系统，其特征在于：其中制冷组件(11)位于所述蠕动泵固定板(8)旁并与所述操作控制面板(55)电性连接。
PCT/CN2021/098551 2021-02-04 2021-06-07 一种全自动wb孵育系统 WO2022166051A1 (zh)

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