CN118010973A - Protein immunoblotting equipment and processing method thereof - Google Patents

Abstract

The invention relates to the technical field of biochemistry, and provides protein immunoblotting equipment which comprises a protein immunoblotting incubation device, a mechanical arm, a liquid adding mechanism, a liquid discharging mechanism, a controller and a constant temperature mechanism. The controller is respectively and electrically connected with the western blotting incubation device, the mechanical arm, the liquid adding mechanism and the constant temperature mechanism. Through the combined work of western blot incubation device, arm, liquid feeding mechanism, flowing back mechanism, controller and constant temperature mechanism, greatly improved the automation of western blot experiment, reduced operating personnel and taken place the error risk in carrying out loaded down with trivial details experimental step. Meanwhile, due to the inclined design of the bottom of the incubation tray in the western blotting incubation device, when the incubation membrane is incubated, the incubation tray only needs to swing at a small angle, so that the risk of liquid splashing in the incubation tray is reduced, and the incubation speed and stability of the incubation membrane are improved.

Description

Protein immunoblotting equipment and processing method thereof

Technical Field

The invention relates to the technical field of biochemistry, in particular to protein immunoblotting equipment and a processing method thereof.

Background

Western blot experiments, i.e., western immunoblotting experiments, are protein analysis techniques that are commonly used to identify certain proteins and to perform qualitative and semi-quantitative analysis of the proteins. In western blotting experiments, the experimental steps involving membrane closure and incubation are extremely critical, and generally, manual assistance is employed to perform experimental operations with semi-automatic equipment.

The semi-automatic device is an antibody incubating and washing shaker capable of saving reagent, which is disclosed in Chinese patent document with publication number of CN 219871356U. The shaking table tray is driven to shake by a driving motor in the driving power bin so as to meet the incubation condition of the incubation film.

In the existing body incubation and cleaning shaker, because of its lower degree of automation, the experimental operator needs to repeatedly carry out a large amount of cleaning and experimental reagent addition work.

Disclosure of Invention

In order to solve the problem of insufficient automation of western blot experimental equipment in the prior art, the invention provides a western blot equipment, which comprises a western blot incubation device, a mechanical arm, a liquid adding mechanism, a liquid discharging mechanism, a controller and a constant temperature mechanism, wherein the controller is respectively electrically connected with the western blot incubation device, the mechanical arm, the liquid adding mechanism and the constant temperature mechanism. By adopting the scheme, through the combined work of the western blotting incubation device, the mechanical arm, the liquid adding mechanism, the liquid discharging mechanism, the controller and the constant temperature mechanism, the automation of the western blotting experiment is greatly improved, and the error risk of operators in the complicated experimental steps is reduced.

Further, the western blot incubation device comprises an incubation disc, a shaking platform, a platform support and a shaking driving mechanism, wherein two ends of the shaking platform are connected with the platform support, the incubation disc is provided with a first containing cavity, the first containing cavity is provided with a first containing cavity top surface positioned at an opening and a first containing cavity bottom surface opposite to the first containing cavity top surface, the first containing cavity bottom surface and a horizontal plane are provided with an included angle alpha, the first containing cavity is further provided with a first side wall, the first side wall on one side of the first containing cavity is a first containing cavity inclined side wall forming an included angle beta with the horizontal plane, one side of the first containing cavity bottom surface, which is close to the first containing cavity inclined side wall, is provided with a blocking column, and the top of the first containing cavity bottom surface is provided with a antislip strip. By adopting the technical scheme, the inclined design of the bottom of the incubation plate in the western blotting equipment can meet the antibody incubation requirement of the incubation membrane under the condition of reducing the use of the first antibody reagent or the second antibody reagent. Meanwhile, due to the inclined design of the bottom of the incubation plate, when the incubation membrane is incubated, the incubation plate only needs to swing at a small angle, so that the risk of liquid splashing in the incubation plate is reduced, and the speed and stability of the incubation membrane incubation work are improved.

Further, shake even actuating mechanism and include first motor, first synchronizing wheel, second synchronizing wheel and hold-in range, first motor is connected with the platform support, and first synchronizing wheel is connected with first motor, and second synchronizing wheel and platform support swivelling joint, and the hold-in range is connected with first synchronizing wheel and second synchronizing wheel.

Further, the mechanical arm comprises an X-axis sliding table module, a Z-axis sliding table module, a Y-axis sliding table module, a cable drag chain, a first water pipe drag chain, a second water pipe drag chain and a third water pipe drag chain.

Z axle slip table module sets up in the output of X axle slip table module, X axle slip table module drive Z axle slip table module along X axle reciprocating motion, Y axle slip table module sets up in the output of Z axle slip table module, Z axle slip table module drive Y axle slip table module along Z axle reciprocating motion, the output of Y axle slip table module is provided with the clamping part in order to centre gripping liquid feeding syringe needle, Y axle slip table module drive clamping part along Y axle reciprocating motion.

The cable drag chain is installed in one side of X axle slip table module and Z axle slip table module, Y axle slip table module, and first water pipe drag chain, second water pipe drag chain and third water pipe drag chain are installed respectively to the opposite side of X axle slip table module and Z axle slip table module, Y axle slip table module.

Further, the filling mechanism includes a peristaltic pump assembly and a first filling container.

The peristaltic pump assembly comprises a peristaltic pump, a fixed support, a second motor and a silicone tube, the peristaltic pump is connected with the fixed support, the peristaltic pump is connected with the second motor, the silicone tube is connected with the peristaltic pump, and one end of the silicone tube sequentially penetrates through the first water tube drag chain, the second water tube drag chain and the third water tube drag chain to be connected with the liquid adding needle.

Further, the liquid draining mechanism comprises a waste liquid groove, a waste liquid collecting runner and a waste liquid collecting container, wherein the waste liquid groove is arranged on one side close to the incubation plate and used for receiving liquid poured from the inclined side wall of the first containing cavity.

Further, the waste liquid tank is provided with a second containing cavity with an upward opening, the second containing cavity is provided with a second containing cavity top surface positioned at the opening and a second containing cavity bottom surface opposite to the second containing cavity top surface, the second containing cavity bottom surface and the horizontal plane are provided with an included angle alpha', the second containing cavity is also provided with a second side wall, and the second side wall of the second containing cavity, which is close to one side of the incubation plate, is a second containing cavity inclined side wall.

The inclined side wall of the second containing cavity comprises a connecting part 51131 and a transition part, the connecting part forms an included angle beta' with the horizontal plane, the cross section of the transition part is in an upward convex arc shape, and the bottom end of the transition part is tangent with the connecting part.

The lowest department in second appearance chamber bottom surface is provided with first through-hole, and first through-hole bottom is provided with first drain pipe, and first drain pipe passes the second through-hole that equipment platform offered to the liquid in the second appearance chamber of discharging.

Further, the waste liquid collecting flow passage is arranged on the bottom surface of the equipment platform, the waste liquid collecting flow passage is arranged below the first drain pipe, a second drain pipe is arranged at the bottom of one end of the waste liquid collecting flow passage, and a water outlet of the second drain pipe penetrates through the top of the second cabinet.

The waste liquid collecting container is arranged below the second drain pipe, and a liquid inlet of the waste liquid collecting container is arranged opposite to a water outlet of the second drain pipe.

Further, the constant temperature mechanism comprises a constant temperature heating fan, a temperature sensor, an outer cover shell and an inner cover shell, wherein the constant temperature heating fan is arranged in a third through hole formed in the equipment platform, the outer cover shell and the inner cover shell are respectively arranged on the top surface of the equipment platform, the inner cover shell and the equipment platform form an operation cabin, and a gap is formed between the outer cover shell and the inner cover shell.

The temperature sensor is respectively arranged above the constant temperature heating fan and below the incubation plate.

The invention also provides a protein immunoblotting treatment method, which comprises the following steps:

s100: the liquid adding mechanism extracts the cleaning liquid to rinse the protein immunoblotting incubation device;

S200: placing an incubation membrane, extracting a first antibody reagent by a liquid adding mechanism, and adding the first antibody reagent into a western blotting incubation device;

s300: the western blot incubation device performs first antibody incubation;

s400: the liquid adding mechanism extracts the cleaning liquid to rinse the incubation film;

s500: the liquid adding mechanism extracts the second antibody reagent and adds the second antibody reagent to the western blotting incubation device;

S600: the western blot incubation device performs second antibody incubation;

S700: the liquid adding mechanism extracts the cleaning liquid to rinse the incubation film;

S800: taking out the incubation film, and extracting the cleaning liquid by a liquid adding mechanism to rinse the western blotting incubation device.

Based on the above, compared with the prior art, the western blotting device provided by the invention can meet the antibody incubation requirement of the incubation membrane under the condition of reducing the use of the first antibody reagent or the second antibody reagent through the inclined design of the bottom of the incubation tray. Meanwhile, due to the inclined design of the bottom of the incubation plate, when the incubation membrane is incubated, the incubation plate only needs to swing at a small angle, so that the risk of liquid splashing in the incubation plate is reduced, and the speed and stability of the incubation membrane incubation work are improved. Through the combined work of western blot incubation device, equipment support, arm, liquid feeding mechanism, flowing back mechanism, controller and constant temperature mechanism, greatly improved the automation of western blot experiment, greatly reduced operating personnel and taken place the error risk in carrying out loaded down with trivial details experimental step.

Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.

Drawings

For a clearer description of embodiments of the invention or of the solutions of the prior art, the drawings that are needed in the description of the embodiments or of the prior art will be briefly described, it being obvious that the drawings in the description below are some embodiments of the invention, and that other drawings can be obtained from them without inventive effort for a person skilled in the art; the positional relationships described in the drawings in the following description are based on the orientation of the elements shown in the drawings unless otherwise specified.

Fig. 1 is a schematic structural diagram of an embodiment of a western blotting apparatus according to the present invention;

fig. 2 is a schematic structural diagram of an embodiment of a western blot incubation device according to the present invention;

FIG. 3 is a schematic structural diagram of an embodiment of a first chamber according to the present invention;

Fig. 4 is a schematic structural diagram II of an embodiment of a western blot incubation device provided by the invention;

FIG. 5 is a schematic structural diagram of an embodiment of a mechanical arm according to the present invention;

fig. 6 is a schematic structural diagram II of an embodiment of a western blotting apparatus according to the present invention;

fig. 7 is a schematic structural diagram III of an embodiment of a western blotting apparatus according to the present invention;

Fig. 8 is a schematic structural diagram of an embodiment of a western blotting apparatus according to the present invention;

FIG. 9 is a flowchart showing steps of a method for processing a protein immunoblot according to the present invention.

Reference numerals:

first holding cavity of incubation tray 111 of 10-protein immunoblotting incubation device 11

1111 First chamber top 1112 first chamber bottom 1113 first chamber sloped sidewall

112 Blocking column 113 anti-slip strip 114 boss

115 Top cover 12 shaking platform 121 buckle

13 Platform support 14 shaking-up driving mechanism 141 first motor

142 First synchronizing wheel 143 second synchronizing wheel 144 synchronous belt

145 Speed reducer 146 idler 147 shading sheet

148 Groove type photoelectric switch 20 equipment support 21 equipment platform

22 First cabinet 221 mounting plate 23 second cabinet

30 Mechanical arm 31X-axis sliding table module 32Z-axis sliding table module

33Y axle slip table module 331 clamping part 332 liquid feeding syringe needle

34 Cable tow chain 35 first water pipe tow chain 36 second water pipe tow chain

37 Third water pipe drag chain 40 liquid adding mechanism 41 peristaltic pump assembly

411 Peristaltic pump 412 silica gel tube 413 fixed support

Second motor 414 first liquid charging container 43 second liquid charging container

50 Liquid discharge mechanism 51 waste liquid tank 511 second cavity

5111 Second chamber top 5112 second chamber bottom 5113 second chamber sloped sidewall

52 Waste liquid collecting channel 521 second drain pipe 53 waste liquid collecting container

60 Controller 70 constant temperature mechanism 71 constant temperature heating fan

72 Temperature sensor 73 outer housing 74 inner housing

75 Working chamber 76 circulation chamber

Detailed Description

For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments of the present invention; the technical features designed in the different embodiments of the invention described below can be combined with each other as long as they do not conflict with each other; all other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

In the description of the present invention, it should be noted that all terms used in the present invention (including technical terms and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs and are not to be construed as limiting the present invention; it will be further understood that terms used herein should be interpreted as having a meaning that is consistent with their meaning in the context of this specification and the relevant art and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein.

Considering that in the existing western blotting experiments, manual assistance is mostly adopted, the experimental efficiency is low, and operators are easy to cause misoperation in a complex experimental flow.

Referring to fig. 1, the present application provides a protein immunoblotting apparatus, which includes a protein immunoblotting incubation device 10, a mechanical arm 30, a liquid feeding mechanism 40, a liquid discharging mechanism 50, a controller 60, and a constant temperature mechanism 70.

The controller 60 is electrically connected with the western blot incubation device 10, the mechanical arm 30, the liquid adding mechanism 40 and the constant temperature mechanism 70 respectively.

According to the western blotting equipment provided by the embodiment, the automation degree of the western blotting equipment is greatly improved through the mutual matching among the western blotting incubation device 10, the mechanical arm 30, the liquid adding mechanism 40, the liquid discharging mechanism 50, the controller 60 and the constant temperature mechanism 70.

In specific implementation, experimental operators only need to manually complete the addition and the removal of the incubation membrane, and can complete the antibody incubation work of the incubation membrane by means of the western blotting equipment.

In order to make the western blotting apparatus in a relatively independent and stable working environment, the western blotting apparatus according to an embodiment of the present application further comprises an apparatus rack 20.

The western blot incubation device 10, the mechanical arm 30, the liquid adding mechanism 40, the liquid discharging system 50, the controller 60 and the constant temperature system 70 are arranged on the equipment bracket 20.

Wherein the equipment rack 20 comprises an equipment platform 21, a first cabinet 22 and a second cabinet 23. The first cabinet 22 and the second cabinet 23 are disposed below the equipment platform 21. The western blot incubation apparatus 10 is mounted on the top surface of the equipment platform 21. The first cabinet 22 is provided with a mounting plate 221 to mount the controller 60.

The controller 60 is electrically connected with the western blot incubation device 10, the mechanical arm 30, the liquid adding mechanism 40 and the constant temperature system 70 respectively.

It will be appreciated that the equipment rack 20 provides a relatively independent and stable working environment for the western blot incubation apparatus 10 and the robotic arm 30, the liquid feeding mechanism 40, the liquid drainage system 50, the controller 60, and the constant temperature system 70.

In order to meet the antibody incubation requirements of the incubation membrane in western blot experiments.

Referring to fig. 2, a western blot incubation apparatus 10 according to an embodiment of the present application includes an incubation tray 11, a shaking platform 12, a platform support 13, and a shaking driving mechanism 14.

In particular, the incubation tray 11 is used for containing an incubation membrane and an experimental reagent, and is used as a carrying container for incubation of the incubation membrane antibody.

Further, incubation plate 11 is mounted on the top surface of shaking platform 12, shaking platform 12 is connected with platform support 13.

In specific implementation, the incubation plate 11 is mounted on the top surface of the shaking platform 12, and the shaking platform 12 is driven to rotate forward or backward by the shaking driving mechanism 14, so as to drive the incubation plate 11 to shake.

Further, the incubation plate 11 is connected to a shaking platform 12.

Preferably, the incubation plate 11 is detachably connected to the shaking platform 12.

The outer wall of the incubation plate 11 is provided with a boss 114, the shaking-up platform 12 is provided with a buckle 121, and the boss 114 is detachably connected with the buckle 121.

Optionally, the incubation plate 11 is detachably connected to the shaking platform 12 by bolts.

In another preferred embodiment, the incubation plate 11 is magnetically coupled to the shake table 12.

Specifically, the bottom of the incubation plate 11 and the top of the shaking platform 12 are respectively provided with a magnetic attraction assembly so as to realize the magnetic attraction connection of the incubation plate 11 and the shaking platform 12.

Further, the shaking-up table 12 is connected to a table support 13.

Preferably, the shaking-up platform 12 is movably connected with the platform support 13.

Specifically, the shaking platform 12 is movably connected with the platform support 13 through rotating shafts arranged at two ends of the shaking platform.

The forward rotation referred to in the present application means that the shaking table 12 rotates clockwise along the rotation shafts provided at both ends of the shaking table 12 near the side where the shaking driving mechanism 14 is provided. Accordingly, the reverse rotation means that the shaking-up platform 12 rotates counterclockwise along the rotating shafts provided at both ends of the shaking-up platform 12 near the side where the shaking-up driving mechanism 14 is provided.

In another embodiment, the top of the incubation plate 11 is provided with a top cover.

During the implementation, the top cap can effectually prevent that the dust in the air from falling into incubation dish 11, prevents to incubate dish 11 and receive the pollution, and then has improved the accuracy of western blot experiment. Meanwhile, the top cover can effectively slow down the evaporation of the liquid in the incubation plate 11, and prevent the change of the reagent concentration during long-time incubation and even cause dry films.

Preferably, the top cover is provided with a through hole.

In the specific implementation, the through hole arranged on the top cover enables the experimental reagent to be added into the incubation plate through the hole when the top cover is in a closed state. Thereby avoid opening and closing of top cap, improve experimental efficiency, reduce pollution risk simultaneously.

Preferably, the top cover is made of transparent materials or is provided with a transparent window.

The top cover made of transparent materials or the top cover is provided with a transparent window, so that experimental operators can observe the experimental reagents and the conditions of the incubation films in the incubation tray 11 at any time.

Referring to fig. 3, the incubation plate 11 of this embodiment has a first cavity 111 with an upward opening.

The first chamber 111 has a first chamber top surface 1111 at the opening and a first chamber bottom surface 1112 opposite the first chamber top surface 1111.

The first chamber bottom 1112 has an angle α with respect to the horizontal plane.

The first cavity bottom surface 1112 forms an angle α with the horizontal plane, such that the first cavity bottom surface 1112 is inclined with respect to the horizontal plane. When the incubation film is placed on the bottom surface 1112 of the first cavity, the incubation plate 11 is shaken to keep the incubation film fully immersed in the experimental reagent under the condition that a small amount of experimental reagent is added. Thereby reducing the usage amount of experimental reagent.

The intersection of the first chamber bottom surface 1112 and the horizontal plane is parallel to the rotation axis of the shaking table 12.

In the design of the first chamber 111 it was found that the angular extent of the angle α has a correlation with the wobble angle of the incubation plate 11. In order to allow the experimental reagent to sufficiently infiltrate the incubation membrane, the larger the angle of the included angle α, the larger the shaking angle of the incubation plate 11 is required. The included angle alpha is half of the shaking angle of the incubation plate 11, so that the incubation requirement of the incubation membrane can be met under the condition that a small amount of experimental reagent is used.

Further, the included angle α is 0 ° -9 °.

In particular, when the angle α is 0 ° -9 °, the inclination angle of the bottom surface 1112 of the first cavity with respect to the horizontal plane is in a reasonable range. The incubation plate 11 is subjected to small-angle shaking so as to increase the shaking frequency and improve the incubation speed of the incubation membrane antibody of the western blotting experiment.

Preferably, the angle α is 3 °.

In particular, when the angle of the included angle α is 3 °, the rocking angle of the incubation plate 11 only needs to be controlled to 6 °. The dosage of the experimental reagent is further reduced, meanwhile, the shaking frequency of the incubation plate 11 is improved, and the incubation speed of the incubation membrane antibody of the western blot experiment is improved.

In one embodiment, the first cavity 111 further has a first sidewall, and the first sidewall on one side of the first cavity 111 is a first cavity inclined sidewall 1113 forming an angle β with the horizontal plane.

The angle beta formed by the first chamber inclined sidewall 1113 and the horizontal plane makes the top of the first chamber inclined sidewall 1113 incline to the outside of the first chamber 111.

The first chamber sloped sidewall 1113 is disposed parallel to the axis of rotation of the shake table 12.

In practice, after the incubation plate 11 rotates to a designed rotation angle toward one side of the first chamber inclined sidewall 1113, the liquid contained in the first chamber 111 is poured out along the first chamber inclined sidewall 1113.

Further, the included angle beta is 15-75 degrees.

In particular, when the angle β is 15 ° -75 °, the inclination angle of the inclined sidewall 1113 of the first chamber with respect to the horizontal plane is within a reasonable range. The pouring of the liquid can be completed by making the incubation plate 11 rotate reversely at a small angle. Preventing slipping of the incubation film caused by an excessive rotation angle of the incubation plate 11.

Referring to fig. 2, the first chamber bottom surface 1112 is provided with an upwardly protruding blocking post 112 near one side of the first chamber sloped sidewall 1113.

Preferably, the blocking post 112 protrudes toward the first chamber top 1111.

In particular, the blocking posts 112 can block the incubation membrane disposed in the incubation tray 11 from sliding out of the incubation tray 11 when the incubation tray 11 is tipped with the assay reagents.

In a preferred embodiment, the blocking post 112 is curved in the direction of the incubation membrane.

In specific implementation, the blocking column 112 is bent towards the direction of the incubation membrane, so that the incubation membrane can be effectively prevented from being separated from the blocking column, and the effect of blocking the membrane from slipping is achieved to a greater extent.

Further, the top of the bottom surface 1112 of the first cavity is provided with an anti-slip strip 113.

In particular, the anti-slip strips 113 increase the roughness of the contact surface between the bottom surface 1112 of the first chamber and the incubation film, and increase the friction between the incubation film and the bottom surface 1112 of the first chamber. To prevent the incubation plate 11 from slipping off the incubation membrane during the incubation or pouring of the liquid.

Preferably, the cross section of the anti-slip strip 113 has an upwardly convex semicircular shape, and the radius of the cross section of the anti-slip strip 113 is 1mm.

Referring to fig. 2, in the present embodiment, the shaking-up driving mechanism 14 includes a first motor 141, a first synchronizing wheel 142, a second synchronizing wheel 143, and a timing belt 144.

In practice, the first synchronous wheel 142 is driven to rotate by the first motor 141. The first synchronizing wheel 141 drives the second synchronizing wheel 143 to rotate through the synchronizing belt 144 so as to drive the shaking platform 12 to rotate forward or backward.

In this embodiment, the first motor 141 is connected to the platform support 13, the first synchronizing wheel 142 is detachably connected to the first motor 141, the second synchronizing wheel 143 is connected to the shaking platform 12, and the timing belt 144 is connected to the first synchronizing wheel 142 and the second synchronizing wheel 143.

Referring to fig. 4, a first motor 141 is detachably connected to the platform support 13, a first synchronizing wheel 142 is detachably connected to the first motor 141, a second synchronizing wheel 143 is fixedly connected to the platform support 13, and a timing belt 144 is connected to the first synchronizing wheel 142 and the second synchronizing wheel 143.

In particular, the first motor 141 is detachably connected to the platform support 13, so that when the first motor 141 fails, an operator can replace it in time.

Preferably, the first motor 141 employs a closed loop stepper motor.

The closed loop stepper motor is a stepper motor with a feedback system that can provide higher control accuracy and reliability than conventional open loop stepper motors.

In particular, the rotation direction and rotation angle of the first synchronous wheel 142 are precisely controlled by the closed-loop stepping motor, so as to achieve the purpose of precisely controlling the shaking angle of the incubation plate 11.

The first synchronizing wheel 142 is detachably connected with the first motor 141 so that an operator can replace damaged parts in time when the first synchronizing wheel 142 or the first motor 141 fails.

Preferably, a decelerator 145 is provided between the first synchronizing wheel 142 and the first motor 141.

In particular, the torque generated by the first motor 141 is amplified by the speed reducer 145 and then transmitted to the first synchronous wheel 142, so as to reduce the power requirement of the first motor 141, which is helpful for saving the motor type selection cost.

The second synchronizing wheel 143 is fixedly connected with the shaking-up platform 12, so as to ensure that the second synchronizing wheel 143 can drive the shaking-up platform 13 to rotate.

Further, an idler wheel 146 is also provided between the first synchronizing wheel 142 and the second synchronizing wheel 143.

In particular, idler wheel 146 is disposed between first and second timing wheels 142, 143 and is coupled to timing belt 144. The pressure angle of the timing belt 144 is adjusted by the idler pulley 146 to optimize the transmission.

Further, the shaking-up platform 12 is provided with a shading sheet 147, and the platform manufacturing 13 is provided with a photoelectric switch 148.

Preferably, the optoelectronic switch 148 is a slot optoelectronic switch.

In particular, the shading sheet 147 rotates in the photoelectric switch 148 to shade light, and when the edge of the shading sheet 147 rotates out of the detection point of the photoelectric switch 148, the photoelectric switch 148 receives a signal, and the position is set as the rotation origin of the shaking platform 12.

Referring to fig. 5, a mechanical arm 30 according to an embodiment of the present application includes an X-axis sliding table module 31, a Z-axis sliding table module 32, a Y-axis sliding table module 33, a cable drag chain 34, a first water pipe drag chain 35, a second water pipe drag chain 36, and a third water pipe drag chain 37.

In specific implementation, the mechanical arm 30 moves in the directions of the X axis, the Z axis and the Y axis through the X axis sliding table module 31, the Z axis sliding table module 32 and the Y axis sliding table module 33. The cables are accommodated by the cable drag chain 34, so that various cables used by the mechanical arm 30 are accommodated and restrained, and cable faults in the operation process of the mechanical arm 30 are prevented. Through first water pipe tow chain 35, second water pipe tow chain 36 and third water pipe tow chain 37, accomodate the constraint to all kinds of water pipes that arm 30 used, prevent that the water pipe from damaging or leading to the jam because of the water pipe is buckled at arm 30 operation in-process.

In this embodiment, the Z-axis sliding table module 32 is disposed at the output end of the X-axis sliding table module 31, and the X-axis sliding table module 31 drives the Z-axis sliding table module 32 to reciprocate along the X-axis. The Y-axis sliding table module 33 is arranged at the output end of the Z-axis sliding table module 32, and the Z-axis sliding table module 32 drives the Y-axis sliding table module 33 to reciprocate along the Z axis. The output end of the Y-axis sliding table module 33 is provided with a clamping part 331 to clamp the liquid adding needle 332, and the Y-axis sliding table module 33 drives the clamping part 331 to reciprocate along the Y axis.

Further, the cable drag chain 34 is installed on one side of the X-axis sliding table module 31, the Z-axis sliding table module 32 and the Y-axis sliding table module 33, and the first water pipe drag chain 35, the second water pipe drag chain 36 and the third water pipe drag chain 37 are respectively installed on the other sides of the X-axis sliding table module 31, the Z-axis sliding table module 32 and the Y-axis sliding table module 33. The X-axis sliding table module 31 is mounted on the top surface of the equipment platform 21.

In specific implementation, the mechanical arm 30 is used for carrying the liquid adding needle 332 arranged at the output end of the Y-axis sliding table module 33 to the upper part of the incubation plate 11 through the matching movement of the X-axis sliding table module 31, the Z-axis sliding table module 32 and the Y-axis sliding table module 33, so as to clean the incubation plate 11 and add experimental reagents.

Referring to fig. 6, an embodiment of the present application provides a priming mechanism 40 comprising a peristaltic pump assembly 41 and a first priming reservoir 42. Peristaltic pump assembly 41 includes peristaltic pump 411, stationary mount 412, second motor 413, and silicone tube 414. Peristaltic pump 411 is connected to a fixed mount 412. Peristaltic pump 411 is coupled to a second motor 413. The silicone tube 414 is connected to peristaltic pump 411. One end of the silicone tube 414 passes through the first water tube drag chain 35, the second water tube drag chain 36 and the third water tube drag chain 37 in sequence and is connected with the liquid adding needle 332.

Further, peristaltic pump 411 is removably coupled to stationary mount 412. Peristaltic pump 411 is movably coupled to a second motor 413. The silicone tube 414 is movably connected with the peristaltic pump 411. One end of the silicone tube 414 sequentially passes through the first water pipe drag chain 35, the second water pipe drag chain 36 and the third water pipe drag chain 37 to be sleeved with the liquid adding needle 332.

In particular, one end of the silicone tube 414 extends into the first liquid adding container 42, and the second motor 413 drives the peristaltic pump 411 to rotate to squeeze the silicone tube 414 installed in the peristaltic pump 411, so as to extract the liquid in the first liquid adding container 42. The liquid pumped from the first liquid charging container 42 is pumped to the liquid charging needle 332 by the peristaltic pump 411 to perform the washing of the incubation plate 11 or the addition of the experimental reagent.

Preferably, the second motor 413 is a closed loop stepper motor.

Specifically, the second motor 413 is a closed-loop stepper motor, and the closed-loop system can monitor the actual position of the motor in real time and ensure that the actual position is consistent with the expected position through an encoder or other position sensor built in the closed-loop stepper motor. And further realize the accurate interpolation of experimental reagent, practice thrift experimental reagent quantity.

Referring to fig. 2, a liquid discharge system 50 according to an embodiment of the present application includes a waste liquid tank 51, a waste liquid collecting flow path 52, and a waste liquid collecting container 53. Waste liquid tank 51 is disposed on a side near incubation plate 11 to receive liquid poured from first chamber sloped sidewall 1113.

In practice, the incubation plate 11 is rotated in the direction of the waste liquid tank 51, and the liquid in the incubation plate 11 is poured. The liquid flows through the waste liquid tank 51 to the waste liquid collecting channel 52, and is collected in the waste liquid collecting container 53.

Further, the waste liquid tank 51 has a second chamber 511 with an upward opening, the second chamber 511 has a second chamber top 5111 at the opening and a second chamber bottom 5112 opposite to the second chamber top 5111, and the second chamber bottom 5112 has an angle α' with respect to the horizontal.

In particular, the included angle α' formed by the second bottom 5112 and the horizontal plane makes the second bottom 5112 incline relative to the horizontal plane. Thereby ensuring that the liquid in the second chamber 511 flows in an inclined direction of the second chamber bottom 5112.

Optionally, the second cavity bottom 5112 may further be provided with a plurality of planes forming different angles with the horizontal plane, and a water outlet is disposed at the lowest position of the adjacent planes.

Optionally, the cross section of the bottom 5112 of the second chamber is in an arc shape protruding upwards, and water outlets are arranged at two ends of the arc shape.

In order to prevent the second chamber 511 from splashing due to a faster flow rate of water when the liquid is poured from the incubation plate 11. The second chamber 511 also has a second side wall. The second side wall of the second chamber 511 near the incubation plate 11 is a second chamber inclined side wall 5113.

The second chamber sloped sidewall 5113 includes a connecting portion 51131 and a transition portion 51132. The connecting portion 51131 forms an angle β' with the horizontal plane. The cross section of the transition part 51132 is in an arc shape protruding upwards. The bottom end of transition 51132 is tangential to connection 51131.

In practice, the incubation plate 11 is rotated in the direction of the waste liquid tank 51, and the liquid in the pouring incubation plate 11 flows along the transition part 51132 to the connection part 51131. Thanks to the arcuate configuration of the transition 51132, the flow rate of liquid poured out of the incubation plate 11 is buffered to avoid the risk of liquid splash.

Further, the included angle beta' is 0 deg. -90 deg..

In particular, when the angle β' is 0 ° -90 °, the inclination angle of the connection portion 51131 with respect to the horizontal plane is in a reasonable range.

Further, a first through hole 512 is formed at the lowest position of the bottom 5112 of the second chamber, a first drain pipe is disposed at the bottom of the first through hole 512, and the first drain pipe passes through a second through hole 211 formed on the equipment platform 21 to drain the liquid 511 in the second chamber.

Referring to fig. 8, the waste liquid collecting channel 52 is attached to the bottom surface of the equipment platform 21. The waste liquid collecting channel 52 is provided below the first drain pipe. The bottom of one end of the waste liquid collecting channel 52 is provided with a second drain pipe 521. The water outlet of the second drain pipe 521 passes through the top of the second cabinet 23. The waste liquid collection container 53 is disposed in the second cabinet 23 below the second drain pipe 521. The liquid inlet of the waste liquid collecting container 53 is opposite to the water outlet of the second drain pipe 521.

In practice, the waste liquid collecting flow path 52 collects the liquid discharged from the waste liquid tank 51 and discharges the collected liquid to the waste liquid collecting container 53 through the second drain pipe 521 to complete the collection of the discharged liquid.

Six kinds of

Referring to fig. 1, 7 and 8, in order to meet the temperature requirement in western blotting experiments, a constant temperature system 70 according to an embodiment of the present application includes a constant temperature heating fan 71, a temperature sensor 72, an outer housing 73 and an inner housing 74.

In particular, the constant temperature heating fan 71

In particular, the inner housing 74 forms a working chamber 75 with the equipment platform 21 providing a closed loop for the incubation tray 11 to be isolated from the environment. The constant temperature heating fan 71 provides a heating source for the operation cabin 75 so as to meet the experimental temperature requirement of western immunoblotting. The constant temperature heating fan 71 serves as an air duct flow output end of the operation cabin 75, and provides cabin air circulation power for the operation cabin 75 so as to uniformly distribute heat in the operation cabin 75.

Further, the constant temperature heating fan 71 is mounted to the third through hole 24 formed in the equipment platform 21. An outer housing 73 and an inner housing 74 are mounted to the top surface of the equipment platform 21, respectively. The inner shroud shell 74 forms a work chamber 75 with the equipment platform 21. A gap is provided between the outer cover housing 73 and the inner cover housing 74.

In practice, the constant temperature heating fan 71 is mounted on one side of the equipment platform 21 to ensure stable air passage in the operation cabin 75. A gap is provided between the outer housing 73 and the inner housing 74 to isolate the operation chamber 75 from heat exchange with the external environment, which helps to maintain the air temperature in the operation chamber 75.

Further, temperature sensors 72 are provided above the constant temperature heating fan 71 and below the incubation tray 11, respectively.

In particular, the temperature of the gas blown out by the constant temperature heating fan 71 is monitored in real time by the temperature sensor 72 to adjust the heating power of the constant temperature heating fan 71. The temperature of the gas under the incubation plate 11 is monitored in real time by the temperature sensor 72 to determine the real time air temperature within the process chamber 75.

Referring to fig. 8, a circulation cabin 76 is provided between the top of the first and second cabinets 22, 23 and the equipment platform 21, and the circulation cabin 76 has side walls. The equipment platform 21 is provided with ventilation holes.

In particular, the circulation chamber 76 serves as a plenum chamber for air circulation within the process chamber 75. The constant temperature heating fan 71 delivers the gas in the circulation chamber 76 to the operation chamber 75, and the gas in the operation chamber 75 flows back to the circulation chamber 76 through the vent hole.

The application also provides a protein immunoblotting treatment method, which comprises the following steps:

s100: the liquid adding mechanism extracts the cleaning liquid to rinse the protein immunoblotting incubation device.

Specifically, the mechanical arm conveys the liquid adding needle to the upper part of an incubation plate in the western blot incubation device, the liquid adding mechanism extracts the cleaning liquid, the incubation plate is moistened by the liquid adding needle, and after the incubation plate is moistened, the western blot incubation device rotates the incubation plate to pour the cleaning liquid.

S200: and (3) placing an incubation membrane, extracting the first antibody reagent by a liquid adding mechanism, and adding the first antibody reagent into a western blotting incubation device.

Specifically, after the incubation film is put into, the mechanical arm conveys the liquid adding needle to the upper part of an incubation disc in the western blotting incubation device, the liquid adding mechanism extracts the first antibody reagent, and a certain amount of the first antibody reagent is added into the incubation disc through the liquid adding needle.

S300: the western blot incubation device performs primary antibody incubation.

Specifically, the western blot incubation device drives the incubation plate to repeatedly shake in the forward and reverse directions, and after incubation of the membrane primary antibody is completed, the western blot incubation device rotates the incubation plate to pour the primary antibody reagent.

S400: the liquid adding mechanism extracts the cleaning liquid to rinse the incubation film.

Specifically, the mechanical arm conveys the liquid adding needle to the upper part of an incubation plate in the western blot incubation device, the liquid adding mechanism extracts the cleaning liquid, the incubation film in the incubation plate is moistened by the liquid adding needle, and after the incubation film is moistened, the western blot incubation device rotates the incubation plate to pour the cleaning liquid.

S500: the liquid adding mechanism extracts the second antibody reagent and adds the second antibody reagent to the western blotting incubation device.

Specifically, the mechanical arm conveys the liquid adding needle to the upper part of an incubation plate in the western blotting incubation device, the liquid adding mechanism extracts the second antibody reagent, and a certain amount of the second antibody reagent is added to the incubation plate through the liquid adding needle.

S600: the western blot incubation device performs a second antibody incubation.

Specifically, the incubation plate in the western blot incubation device is repeatedly shaken in the forward and reverse directions, and after incubation of the second antibody of the incubation membrane is completed, the western blot incubation device rotates the incubation plate to pour the second antibody reagent.

S700: the liquid adding mechanism extracts the cleaning liquid to rinse the incubation film.

Specifically, the mechanical arm conveys the liquid adding needle to the upper part of an incubation plate in the western blot incubation device, the liquid adding mechanism extracts the cleaning liquid, the incubation film in the incubation plate is moistened by the liquid adding needle, and after the incubation film is moistened, the western blot incubation device rotates the incubation plate to pour the cleaning liquid.

S800: taking out the incubation film, and extracting the cleaning liquid by a liquid adding mechanism to rinse the western blotting incubation device.

Specifically, after taking out the incubation membrane, the mechanical arm conveys the liquid adding needle to the upper part of an incubation disc in the western blotting incubation device, and after the liquid adding mechanism extracts the cleaning liquid to moisten the incubation disc, the western blotting incubation device rotates to pour the cleaning liquid.

According to the protein immunoblotting processing method provided by the application, the manual workload in the protein immunoblotting experiment is reduced through the combined work of the protein immunoblotting incubation device, the mechanical arm, the liquid adding mechanism, the liquid discharging mechanism, the controller and the constant temperature mechanism. The experimental efficiency is improved, and the accuracy of experimental results is improved.

In summary, compared with the prior art, the western blotting equipment provided by the invention has the advantages that the automation of the western blotting experiment is greatly improved and the error risk of operators in complex experimental steps is reduced through the combined work of the western blotting incubation device, the mechanical arm, the liquid adding mechanism, the liquid draining mechanism, the controller and the constant temperature mechanism.

The inclined design of the bottom of the incubation tray in the western blotting equipment can meet the antibody incubation requirement of the incubation membrane under the condition of reducing the use of the first antibody reagent or the second antibody reagent. Meanwhile, due to the inclined design of the bottom of the incubation plate, when the incubation membrane is incubated, the incubation plate only needs to swing at a small angle, so that the risk of liquid splashing in the incubation plate is reduced, and the speed and stability of the incubation membrane incubation work are improved.

In addition, it should be understood by those skilled in the art that although there are many problems in the prior art, each embodiment or technical solution of the present invention may be modified in only one or several respects, without having to solve all technical problems listed in the prior art or the background art at the same time. Those skilled in the art will understand that nothing in one claim should be taken as a limitation on that claim.

Although terms such as western blot incubation apparatus, equipment rack, robotic arm, liquid feeding mechanism, liquid draining mechanism, controller, thermostatic mechanism, etc. are used more herein, the possibility of using other terms is not excluded. These terms are used merely for convenience in describing and explaining the nature of the invention; they are to be interpreted as any additional limitation that is not inconsistent with the spirit of the present invention; the terms first, second, and the like in the description and in the claims of embodiments of the invention and in the above-described figures, if any, are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.

Claims (10)

1. Protein immunoblotting equipment, its characterized in that: including western blot incubation device (10), arm (30), liquid feeding mechanism (40), flowing back mechanism (50), controller (60) and constant temperature mechanism (70), controller (60) respectively with western blot incubation device (10) with arm (30) liquid feeding mechanism (40) constant temperature mechanism (70) electricity is connected.

2. The western blot apparatus of claim 1, wherein: the western blot incubation device (10) comprises an incubation disc (11), a shaking platform (12), a platform support (13) and a shaking driving mechanism (14), wherein two ends of the shaking platform (12) are connected with the platform support (13), the incubation disc (11) is provided with a first accommodating cavity (111), the first accommodating cavity (111) is provided with a first accommodating cavity top surface (1111) positioned at an opening and a first accommodating cavity bottom surface (1112) opposite to the first accommodating cavity top surface (1111), the first accommodating cavity bottom surface (1112) is provided with an included angle alpha with a horizontal plane, the first accommodating cavity (111) is further provided with a first side wall, and the first side wall on one side of the first accommodating cavity (111) is a first accommodating cavity inclined side wall (1113) forming an included angle beta with the horizontal plane.

3. The western blot apparatus of claim 2, wherein: the shaking-up driving mechanism (14) comprises a first motor (141), a first synchronizing wheel (142), a second synchronizing wheel (143) and a synchronous belt (144), wherein the first motor (141) is connected with the platform support (13), the first synchronizing wheel (142) is connected with the first motor (141), the second synchronizing wheel (143) is in rotary connection with the platform support (13), and the synchronous belt (144) is connected with the first synchronizing wheel (142) and the second synchronizing wheel (143).

4. The western blot apparatus of claim 1, wherein: the mechanical arm (30) comprises an X-axis sliding table module (31), a Z-axis sliding table module (32), a Y-axis sliding table module (33), a cable drag chain (34), a first water pipe drag chain (35), a second water pipe drag chain (36) and a third water pipe drag chain (37);

the Z-axis sliding table module (32) is arranged at the output end of the X-axis sliding table module (31), the X-axis sliding table module (31) drives the Z-axis sliding table module (32) to reciprocate along the X-axis, the Y-axis sliding table module (33) is arranged at the output end of the Z-axis sliding table module (32), the Z-axis sliding table module (32) drives the Y-axis sliding table module (33) to reciprocate along the Z-axis, the output end of the Y-axis sliding table module (33) is provided with a clamping part (331) for clamping a liquid feeding needle head (332), and the Y-axis sliding table module (33) drives the clamping part (331) to reciprocate along the Y-axis;

the cable drag chain (34) is installed in X axle slip table module (31) with Z axle slip table module (32) one side of Y axle slip table module (33), X axle slip table module (31) with Z axle slip table module (32) the opposite side of Y axle slip table module (33) is installed respectively first water pipe drag chain (35), second water pipe drag chain (36) with third water pipe drag chain (37).

5. The western blot apparatus of claim 1, wherein: the liquid adding mechanism (40) comprises a peristaltic pump assembly (41) and a first liquid adding container (42);

peristaltic pump subassembly (41) are including peristaltic pump (411), fixed bolster (412), second motor (413) and silicone tube (414), peristaltic pump (411) with fixed bolster (412) are connected, peristaltic pump (411) with second motor (413) are connected, silicone tube (414) with peristaltic pump (411) are connected, one end of silicone tube (414) passes in proper order first water pipe tow chain (35) second water pipe tow chain (36) and third water pipe tow chain (37) with liquid feeding syringe needle (332) are connected.

6. The western blot apparatus of claim 1, wherein: the liquid draining mechanism (50) comprises a waste liquid groove (51), a waste liquid collecting flow channel (52) and a waste liquid collecting container (53), wherein the waste liquid groove (51) is arranged on one side close to the incubation plate (11) so as to receive liquid poured from the inclined side wall (1113) of the first containing cavity.

7. The western blot apparatus of claim 1, wherein: the waste liquid tank (51) is provided with a second containing cavity (511) with an upward opening, the second containing cavity (511) is provided with a second containing cavity top surface (5111) positioned at the opening and a second containing cavity bottom surface (5112) opposite to the second containing cavity top surface (5111), the second containing cavity bottom surface (5112) and the horizontal plane are provided with an included angle alpha', the second containing cavity (511) is also provided with a second side wall, and the second side wall of the second containing cavity (511) close to one side of the incubation plate (11) is a second containing cavity inclined side wall (5113);

The second accommodating cavity inclined side wall (5113) comprises a connecting part (51131) and a transition part (51132), an included angle beta' is formed between the connecting part (51131) and the horizontal plane, the cross section of the transition part (51132) is in an upward convex arc shape, and the bottom end of the transition part (51132) is tangent to the connecting part (51131);

the bottom of the second containing cavity (5112) is provided with a first through hole (512), the bottom of the first through hole (512) is provided with a first drain pipe, and the first drain pipe penetrates through a second through hole (211) formed in the equipment platform (21) so as to discharge liquid in the second containing cavity (511).

8. The western blot apparatus of claim 7, wherein: the waste liquid collecting flow passage (52) is arranged on the bottom surface of the equipment platform (21), the waste liquid collecting flow passage (52) is arranged below the first drain pipe, a second drain pipe (521) is arranged at the bottom of one end of the waste liquid collecting flow passage (52), and a water outlet of the second drain pipe (521) penetrates through the top of the second cabinet (23);

the waste liquid collecting container (53) is arranged below the second drain pipe (521), and a liquid inlet of the waste liquid collecting container (53) is opposite to a water outlet of the second drain pipe (521).

9. The western blot apparatus of claim 1, wherein: the constant temperature mechanism (70) comprises a constant temperature heating fan (71), a temperature sensor (72), an outer cover shell (73) and an inner cover shell (74), wherein the constant temperature heating fan (71) is installed on a third through hole (24) formed in the equipment platform (21), the outer cover shell (73) and the inner cover shell (74) are respectively installed on the top surface of the equipment platform (21), the inner cover shell (74) and the equipment platform (21) form a working cabin (75), and a gap is formed between the outer cover shell (73) and the inner cover shell (74);

The temperature sensors (72) are respectively arranged above the constant temperature heating fan (71) and below the incubation plate (11).

10. A protein immunoblotting processing method is characterized by comprising the following steps:

s100: the liquid adding mechanism extracts the cleaning liquid to rinse the protein immunoblotting incubation device;

S200: placing an incubation membrane, extracting a first antibody reagent by a liquid adding mechanism, and adding the first antibody reagent into a western blotting incubation device;

s300: the western blot incubation device performs first antibody incubation;

s400: the liquid adding mechanism extracts the cleaning liquid to rinse the incubation film;

s500: the liquid adding mechanism extracts the second antibody reagent and adds the second antibody reagent to the western blotting incubation device;

S600: the western blot incubation device performs second antibody incubation;

S700: the liquid adding mechanism extracts the cleaning liquid to rinse the incubation film;

S800: taking out the incubation film, and extracting the cleaning liquid by a liquid adding mechanism to rinse the western blotting incubation device.