High-throughput western blot incubation equipment and method

Abstract

The invention discloses high-flux western blot incubation equipment and method.The high-flux western blot incubation equipment comprises incubation plates, a liquid adding assembly and a recycling assembly, the incubation plates are connected through a support assembly and used for containing incubation membranes to be incubated, and the support assembly is provided with an upper bearing part, a middle bearing part and a lower bearing part which are sequentially arranged from top to bottom; the incubation plate is arranged on the middle bearing part; the liquid adding assembly is arranged on the upper bearing part of the bracket assembly through the first driving assembly and is used for driving the liquid adding assembly to inject quantitative incubation liquid into the incubation disc through the first driving assembly; the recovery assembly is arranged on the lower bearing part of the bracket assembly and is used for recovering the liquid in the incubation tray; wherein the liquid adding assembly is provided with at least a first storage area and a second storage area, and the first storage area and the second storage area are distributed oppositely and are used for storing different types of incubation liquid so as to add the incubation liquid to the incubation plate. By means of the structural design, the incubation automation degree is greatly improved, and meanwhile the multi-step incubation operation time is shortened.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biochemical technology, more specifically, relates to a high-throughput protein immunoblotting incubation device and method. BACKGROUND

[0002] Western Blot is the core experiment in molecular biology, clinical diagnosis and drug discovery, which can detect dozens to hundreds of target proteins at a time. In the past decade, with the rapid growth of multi-target drug screening, tumor marker panels and personalized treatment needs, laboratories have upgraded the traditional single-target "manual film" to high-throughput mode of 96-well or even 384-well plates, and the daily sample processing capacity has jumped from dozens to thousands. However, high throughput does not mean high intelligence. The so-called "high-throughput Western Blot instrument" on the market is mostly a simple assembly of mechanical arms, peristaltic pumps and imaging modules, and it still belongs to the "semi-automatic" category in essence.

[0003] Under this semi-automatic framework, each batch still needs to be manually completed by the experimenter for the key steps of adding primary antibody, adding secondary antibody, washing the membrane and developing color, which not only consumes a lot of manpower, but also alternates between "waiting for the machine" and "waiting for the person", resulting in a whole throughput much lower than the theoretical value. More seriously, manual intervention tears the strict time window into pieces: the incubation time may differ by several minutes or even half an hour between different wells, and the antibody binding kinetics and signal amplification efficiency drift, resulting in significant batch-to-batch differences in band intensity and difficulty in experimental replication, which directly affects downstream quantitative analysis and clinical interpretation.

[0004] In addition, reagents such as antibodies and chemiluminescent substrates are often added in microliter level, and the price is comparable to that of gold by volume; the open liquid path design of the semi-automatic system makes it impossible to recover residual liquid. At the same time, the multi-channel sample addition needle is not thoroughly cleaned when switching between different antibodies or blocking liquids, and cross contamination easily occurs, which may result in high background, specific signal being overwhelmed, or even the whole plate being scrapped. Therefore, how to break through in the four dimensions of "full automation, time synchronization, reagent recovery, and zero cross contamination" at the same time has become a bottleneck restricting the large-scale application of high-throughput Western Blot. SUMMARY

[0005] In view of the above defects or improvement needs of the prior art, the present application significantly improves the operation efficiency and automation level by integrating the layered support assembly (upper, middle and lower bearing parts) with the functional assembly (liquid addition, incubation and recovery). Specifically, in a first aspect, the present application provides a high-throughput protein immunoblotting incubation device, comprising an incubation disc, a liquid addition assembly and a recovery assembly. The incubation tray is connected by a support assembly for containing an incubation membrane to be incubated, the support assembly has an upper bearing part, a middle bearing part and a lower bearing part arranged in sequence from top to bottom, and the incubation tray is arranged in the middle bearing part; The liquid adding assembly is arranged in the upper bearing part of the support assembly by a first driving assembly, and is used for adding a certain amount of incubation liquid into the incubation tray by driving the liquid adding assembly by the first driving assembly; The recovery assembly is arranged in the lower bearing part of the support assembly, and is used for recovering the liquid in the incubation tray; The liquid adding assembly has at least a first storage area and a second storage area, and the first storage area and the second storage area are oppositely arranged for storing different kinds of incubation liquid to add to the incubation tray.

[0006] In the first aspect, the first storage area and the second storage area of the liquid adding assembly are respectively provided with a first guide part and a second guide part corresponding to the side parts away from each other, and the first guide part or the second guide part is used for guiding the incubation liquid to flow naturally along the respective guide direction; The incubation tray is open at the upper part, and the volume of the upper opening is greater than the area of the liquid adding assembly; The first driving assembly is used for driving the liquid adding assembly to tilt towards the first storage area or the second storage area, when the first driving assembly drives the liquid adding assembly to tilt towards the first storage area, the incubation liquid in the first storage area flows naturally along the guide direction of the first guide part to the incubation tray, and when the first driving assembly drives the liquid adding assembly to tilt towards the second storage area, the incubation liquid in the first storage area flows naturally along the guide direction of the second guide part to the incubation tray.

[0007] In the first aspect, the liquid adding assembly includes a liquid adding groove, the middle part of the liquid adding groove is divided into the first storage area and the second storage area by a partition, and the bottom of the liquid adding groove is further provided with a chute; The upper bearing part is further provided with a bracket, and the chute is slidingly arranged in the bracket and can slide along the length direction of the bracket.

[0008] In the first aspect, the middle part of the liquid adding groove is further provided with a liquid adding avoiding hole, the liquid adding avoiding hole is a cylindrical structure formed by surrounding, has an inlet and an outlet, and is located between the first storage area and the second storage area; The inlet is flush with the groove of the liquid adding groove, and the outlet faces the incubation tray.

[0009] In the first aspect, the incubation tray is arranged on the middle bearing part of the support assembly by a second driving mechanism, and is driven to tilt left and right by the second driving mechanism to shake the incubation liquid in the incubation tray.

[0010] In the first aspect, the incubation tray has a third guide part and a fourth guide part, which are oppositely arranged on the incubation tray. The upper tray opening of the incubation tray is provided with a cover, and the cover is provided with a first liquid inlet, a second liquid inlet and a third liquid inlet. The first liquid inlet and the second liquid inlet are oppositely arranged, and the first liquid inlet and the second liquid inlet are located directly below the first guide part and the second guide part of the liquid adding assembly, respectively, for receiving the incubation liquid guided by the first guide part or the second guide part. The third liquid inlet is opposite to the outlet of the liquid adding avoiding hole.

[0011] In the first aspect, the recovery assembly includes a first recovery part and a second recovery part, which are located directly below the third guide part and the fourth guide part, respectively. The first recovery part includes a waste liquid recovery tank, and the second recovery part includes an incubation liquid recovery pipe.

[0012] In the first aspect, the first guide part, the second guide part, the third guide part and the fourth guide part are all conical.

[0013] In the first aspect, the liquid adding assembly is arranged above the support assembly and above the liquid adding assembly, and is used to supplement the incubation liquid to the first storage area and the second storage area of the liquid adding assembly. The liquid adding assembly includes a first liquid adding support, a second liquid adding support and a turnover assembly. The first liquid adding support and the second liquid adding support are arranged vertically, and the turnover assembly is used to drive the first liquid adding support and the second liquid adding support to turn left and right. The first liquid adding support and the second liquid adding support are fixedly provided with centrifugal tubes for filling incubation liquid. When any one of the first liquid adding support and the second liquid adding support is turned to any one of the left side and the right side, the incubation liquid in the centrifugal tube on the corresponding side is poured into the first storage area or the second storage area, and the centrifugal tube on the first liquid adding support or the second liquid adding support which is not turned to any one of the left side and the right side remains with the tube body and the tube opening upward.

[0014] In the second aspect, the present application provides a high-throughput protein immunoblotting incubation method, which is applied to a high-throughput protein immunoblotting incubation device. The method includes the following steps: Step S1, the first time extraction of cleaning solution is used to clean the incubation disc through the liquid adding avoidance hole, and then the cleaning solution is introduced into the waste liquid recovery groove through the second driving mechanism to be recovered; Step S2, the incubation membrane is put in, the first antibody reagent is extracted into the first storage area of the liquid adding assembly, and the first storage area of the liquid adding assembly is driven to be inclined to the incubation disc direction through the first driving assembly, so that the first antibody reagent is introduced into the incubation disc; Step S3, the incubation disc is incubated with the first antibody; Step S4, the second time extraction of cleaning solution is used to clean the incubation disc through the liquid adding avoidance hole, and then the cleaning solution is introduced into the waste liquid recovery groove through the second driving mechanism to be recovered; Step S5, the second antibody reagent is extracted into the second storage area of the liquid adding assembly, and the second storage area of the liquid adding assembly is driven to be inclined to the incubation disc direction through the first driving assembly, so that the second antibody reagent is introduced into the incubation disc; Step S6, the incubation disc is incubated with the second antibody; Step S7, the incubation membrane is taken out, the third time extraction of cleaning solution is used to clean the incubation disc through the liquid adding avoidance hole, and then the cleaning solution is introduced into the waste liquid recovery groove through the second driving mechanism to be recovered.

[0015] Overall, compared with the prior art, the above technical solutions conceived by the present application can achieve the following beneficial effects: 1. The high-throughput protein immunoblotting incubation device can significantly improve the operation efficiency and automation level through the integrated design of the layered support assembly (upper, middle and lower bearing parts) and the functional assembly (liquid adding, incubation and recovery). The incubation disc is fixed to the middle bearing part, which is convenient for stable bearing. The liquid adding assembly is arranged in the upper bearing part through the driving mechanism, which can accurately add liquid to the lower incubation disc. The recovery assembly is located in the lower bearing part, which can directly receive and recover the waste liquid flowing out of the incubation disc. This vertical space layout realizes smooth connection and space isolation of the liquid adding, incubation and recovery processes, effectively avoids liquid dripping cross contamination, reduces manual intervention links, and significantly improves experimental throughput and operation convenience.

[0016] 2、The liquid adding assembly adopts a first storage area and a second storage area designed in a standing type, and combined with the quantitative control of the first driving assembly, the high-throughput processing capability and experimental consistency are effectively ensured. The design allows the device to store and conveniently switch between two different incubation liquids at the same time; for example, primary antibodies, secondary antibodies, washing liquids, etc. Without the need to frequently replace or clean the liquid adding unit, the time of multi-step incubation operation is greatly shortened. The first driving assembly ensures that each time the injection is a precise set amount, which not only reduces the waste of expensive reagents, but more importantly, ensures the uniformity of the incubation conditions of the same batch or different batches of samples, providing key support for the reliability and repeatability of high-throughput Western blotting experimental results. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 Structure diagram of a high-throughput Western blotting incubation device in an embodiment of the present application Figure 1 ; Figure 2 Structure diagram of a high-throughput Western blotting incubation device in an embodiment of the present application Figure 2 ; Figure 3 Structure diagram of a high-throughput Western blotting incubation device in an embodiment of the present application Figure 3 ; Figure 4 Structure diagram of a high-throughput Western blotting incubation device in an embodiment of the present application Figure 4 ; Figure 5 Structure diagram of a high-throughput Western blotting incubation device in an embodiment of the present application Figure 5 ; Figure 6 Structure diagram of a high-throughput Western blotting incubation device in an embodiment of the present application Figure 6 ; Figure 7 Flowchart of a high-throughput Western blotting incubation method in an embodiment of the present application.

[0019] 1, incubation tray; 101, third guide part; 102, fourth guide part; 103, cover; 10301, first liquid inlet; 10302, second liquid inlet; 10303, third liquid inlet; 2, liquid adding assembly; 201, first storage area; 20101, first guide part; 202, second storage area; 20201, second guide part; 203, liquid adding avoidance hole; 3, recovery assembly; 301, waste liquid recovery tank; 302, incubation liquid recovery pipe; 4, support assembly; 401, upper bearing part; 402, middle bearing part; 403, lower bearing part; 5. first driving assembly; 501. stepper motor; 502. synchronous pulley assembly; 6. bracket; 7. second driving assembly; 8. liquid feeding assembly; 801. first liquid feeding bracket; 802. second liquid feeding bracket; 803. overturning assembly; 9. centrifugal tube. DETAILED DESCRIPTION

[0020] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and should not be used to limit the present application. In addition, the technical features involved in the various embodiments of the present application described below can be combined with each other as long as they do not conflict with each other.

[0021] Embodiment one: Please refer to Figures 1-6 The present embodiment one provides a high-throughput protein immunoblotting incubation device, which comprises an incubation tray 1, a liquid feeding assembly 2 and a recovery assembly 3. The incubation tray 1 is connected by a support assembly 4 for containing the incubation membrane to be incubated. The support assembly 4 has an upper bearing part 401, a middle bearing part 402 and a lower bearing part 403 arranged in sequence from top to bottom. The incubation tray 1 is arranged in the middle bearing part 402. The liquid feeding assembly 2 is arranged in the upper bearing part 401 of the support assembly 4 by a first driving assembly 5. The first driving assembly 5 drives the liquid feeding assembly 2 to inject a certain amount of incubation liquid into the incubation tray 1. The recovery assembly 3 is arranged in the lower bearing part 403 of the support assembly 4 for recovering the liquid in the incubation tray 1. The liquid feeding assembly 2 has at least a first storage area and a second storage area. The first storage area and the second storage area are distributed in an opposite manner for storing different kinds of incubation liquid to be injected into the incubation tray 1.

[0022] Specifically, the high-throughput protein immunoblotting incubation device of the present application significantly improves the operation efficiency and automation level through the integrated design of the upper, middle and lower bearing parts 403 of the layered support assembly 4 and the liquid adding, incubation and recovery functional assemblies. The incubation plate 1 is fixed to the middle bearing part 402, which facilitates stable bearing; the liquid adding assembly 2 is arranged on the upper bearing part 401 through a driving mechanism, which can accurately add liquid to the lower incubation plate 1; and the recovery assembly 3 is located on the lower bearing part 403, which can directly receive and recover the waste liquid flowing out of the incubation plate 1. This vertical spatial layout realizes smooth connection and spatial isolation of the liquid adding, incubation and recovery processes, effectively avoids liquid dripping and cross contamination, reduces manual intervention, and significantly improves experimental throughput and operation convenience. The liquid adding assembly 2 adopts a design of opposite first and second storage areas, which, in combination with the quantitative control of the first driving assembly 5, effectively guarantees high-throughput processing capacity and experimental consistency. This design allows the device to store and conveniently switch between adding two different incubation liquids, such as primary antibodies, secondary antibodies and washing liquids, without the need for frequent replacement or cleaning of the liquid adding unit, greatly shortening the time of multi-step incubation operation. The first driving assembly 5 ensures that the amount of liquid added each time is accurately set, which not only reduces the waste of expensive reagents, but more importantly, guarantees the uniformity of incubation conditions for the same batch or different batches of samples, providing key support for the reliability and repeatability of high-throughput protein immunoblotting experimental results.

[0023] Please refer to Figure 3 In a specific embodiment, the first storage area 201 and the second storage area 202 of the liquid adding assembly 2 are respectively provided with a first guide part 20101 and a second guide part 20201 corresponding to the side parts thereof, and the first guide part 20101 or the second guide part 20201 is used to guide the natural flow of the incubation liquid in the respective guide direction; the incubation plate 1 is in the form of an upper opening, and the volume of the upper opening is greater than the area of the liquid adding assembly 2; wherein the first driving assembly 5 is used to drive the liquid adding assembly 2 to tilt towards the first storage area 201 or the second storage area 202, when the first driving assembly 5 drives the liquid adding assembly 2 to tilt towards the first storage area 201, the incubation liquid in the first storage area 201 naturally flows to the incubation plate 1 along the guide direction of the first guide part 20101, and when the first driving assembly 5 drives the liquid adding assembly 2 to tilt towards the second storage area 202, the incubation liquid in the first storage area 201 naturally flows to the incubation plate 1 along the guide direction of the second guide part 20201.

[0024] Specifically, by arranging the first guide part 20101 and the second guide part 20201 on the mutually distal sides of the first storage area 201 and the second storage area 202, and enabling the liquid adding assembly 2 as a whole to produce controllable tilting under the action of the first driving assembly 5, the different incubation liquids can be naturally flowed into the incubation tray 1 along the respective independent guide directions only by relying on gravity, without the need for shared pumps, valves or pipelines, thereby avoiding cross contamination and reducing system complexity. The upper opening area of the incubation tray 1 is greater than the projection area of the liquid adding assembly 2, which ensures that the liquid can be completely received without splashing out during tilting drainage; in combination with the single degree of freedom action of tilting-back, the cycle of liquid adding-stopping-switching can be completed, which simplifies the driving mechanism and improves the reliability of liquid adding positioning.

[0025] Please continue to refer to Figure 3 In a specific embodiment, the liquid adding assembly 2 comprises a liquid adding groove, the middle part of the liquid adding groove is divided into the first storage area and the second storage area 202 by a partition, and the volumes of the two areas are equal, and the inside of the first storage area or the second storage area 202 is arranged in a stepped shape from the bottom to the top, and the bottom of the liquid adding groove is further provided with a sliding groove; The upper bearing part 401 is further provided with a bracket 6, the sliding groove is slidingly arranged in the bracket 6 and can slide along the length direction of the bracket 6, and the bracket 6 is further provided with a clamping groove which is detachably connected with the bottom of the liquid adding groove.

[0026] In a specific embodiment, the middle part of the liquid adding groove is further provided with a liquid adding avoiding hole 203, the liquid adding avoiding hole 203 is a cylindrical structure formed by surrounding, has an inlet and an outlet, and is located between the first storage area and the second storage area 202; wherein the inlet is flush with the slot of the liquid adding groove, and the outlet is opposite to the incubation tray 1, so as to add cleaning liquid through the inlet and guide the cleaning liquid into the incubation tray 1 through the outlet to clean the incubation tray 1.

[0027] In a specific embodiment, the incubation tray 1 is arranged on the middle bearing part 402 of the bracket assembly 4 through the second driving assembly 7, for driving the incubation tray 1 to tilt left and right by the second driving assembly 7, so as to shake the incubation liquid in the incubation tray 1.

[0028] Please refer to Figures 4-5 In a specific embodiment, the incubation tray 1 has a third guide part 101 and a fourth guide part 102, the third guide part 101 and the fourth guide part 102 are distributed in an opposite manner on the incubation tray 1. The upper disc opening of the incubation disc 1 is provided with a cover 103, and the cover 103 is provided with a first liquid inlet 10301, a second liquid inlet 10302 and a third liquid inlet 10303. The first liquid inlet 10301 and the second liquid inlet 10302 are oppositely arranged, and the first liquid inlet 10301 and the second liquid inlet 10302 are located directly below the first guide part 20101 and the second guide part 20201 of the liquid adding assembly 2 respectively, for receiving the incubation liquid guided by the first guide part 20101 or the second guide part 20201. The third liquid inlet 10303 is directly opposite the outlet of the liquid adding avoiding hole 203. The partitioned flow guide of the incubation disc 1 is cooperated with the multiple liquid inlets of the cover 103, which significantly optimizes the directional delivery efficiency of reagents and avoids cross contamination. Through the cooperation of the third and fourth opposite guide parts provided on the disc body and the first and second liquid inlets 10302 and the third liquid inlet 10303 specially arranged on the cover 103, the precise partitioned injection and flow guide of different types of incubation liquid are realized. The first and second liquid inlets 10302 respectively receive reagents from different storage areas of the liquid adding assembly 2, ensuring independent delivery paths for sensitive reagents such as antibodies and blocking solution. The third liquid inlet 10303 is dedicated to the addition of auxiliary liquids such as buffer solution. This structure not only avoids the mixing risk of multiple reagents during the addition process, but also effectively prevents cross contamination of expensive reagents through physical isolation, thereby improving the reliability of experimental results. The recovery assembly 3 includes a first recovery part and a second recovery part, and the first recovery part and the second recovery part are located directly below the third guide part 101 and the fourth guide part 102 respectively. The first recovery part includes a waste liquid recovery tank 301, and the second recovery part includes an incubation liquid recovery pipe 203. The waste liquid recovery tank 301 of the first recovery part and the incubation liquid recovery pipe 203 of the second recovery part are respectively arranged corresponding to the third and fourth guide parts 102 of the incubation disc 1, forming independent recovery paths. This design enables automatic classified recovery of waste liquids such as washing waste liquid and reusable uncontaminated incubation liquids such as excess antibody solution: waste liquid enters the recovery tank for centralized treatment, while uncontaminated valuable reagents are safely collected through a dedicated recovery pipe for reuse. This not only reduces the cost of experimental consumables, but also reduces the discharge of harmful waste liquid, achieving resource conservation and environmentally friendly operation during the experimental process.

[0029] In a specific embodiment, the first guide part 20101, the second guide part 20201, the third guide part 101 and the fourth guide part 102 are all conical, which facilitates directional guidance of the liquid.

[0030] Please refer to Figure 6In a specific embodiment, the upper liquid component 8 is arranged on the support component 4 and above the liquid adding component 2, and is used to supplement the first storage area and the second storage area of the liquid incubation component 2 with the incubation liquid; the upper liquid component 8 comprises a first upper liquid support 801, a second upper liquid support 802, and a turnover component 803, the first upper liquid support 801 is arranged vertically with the second upper liquid support 802, and the turnover component 803 is used to drive the first upper liquid support 801 and the second upper liquid support 802 to turn left and right; through the cooperation of the vertically arranged first upper liquid support 801, the second upper liquid support 802, and the turnover component 803, the device can automatically supplement different kinds of reagents without interrupting the incubation process: when one side of the upper liquid support turns to accurately pour the reagent in the centrifugal tube 9 into the corresponding storage area, the other side of the upper liquid support always maintains the posture of the tube opening upward, ensuring that there is no risk of leakage of the standby reagent. This design not only completely replaces the manual opening and liquid supplementing operation, avoids the experimental interruption and pollution risk caused by frequent intervention, but also ensures zero waste of valuable reagents through the precise control of mechanical turning. The first upper liquid support 801 and the second upper liquid support 802 are fixedly provided with centrifugal tubes 9 for filling incubation liquid, and when any one of the first upper liquid support 801 or the second upper liquid support 802 is turned to either side, the incubation liquid in the centrifugal tube 9 on the corresponding side is poured into the first storage area or the second storage area, and the centrifugal tube 9 on the first upper liquid support 801 or the second upper liquid support 802 which is not on either side keeps the tube body and the tube opening upward. Specifically, the turnover component 803 drives the double upper liquid supports to switch between left and right stations, forming a cycle mode of "one side pouring and supplementing, the other side preparing standby". When the current sequence reagent is about to run out, the system can instantaneously switch to the standby upper liquid support for liquid supplementing, ensuring that the first storage area and the second storage area are always in a full state. This seamless replenishment mode breaks through the efficiency bottleneck caused by liquid supplementing downtime in traditional devices, and is especially suitable for large-scale, multi-batch high-throughput Western blot experiments, providing stable automation support for ultra-long time or multi-round incubation.

[0031] Embodiment two: Please refer to Figure 7The embodiment two provides a high-throughput protein immunoblotting incubation method, the method is applied to a high-throughput protein immunoblotting incubation device, and the method comprises the following steps: S1, first extracting cleaning solution to clean the incubation disc through the liquid adding avoidance hole, then the incubation disc is inclined through the second driving mechanism to guide the cleaning solution into the waste liquid recovery groove for recovery; S2, the incubation membrane is placed, the first antibody reagent is extracted to the first storage area of the liquid adding assembly, and the first storage area of the liquid adding assembly is inclined to the direction of the incubation disc through the first driving assembly to guide the first antibody reagent into the incubation disc; S3, the incubation disc is incubated with the first antibody; S4, second extracting cleaning solution to clean the incubation disc through the liquid adding avoidance hole, then the incubation disc is inclined through the second driving mechanism to guide the cleaning solution into the waste liquid recovery groove for recovery; S5, the second antibody reagent is extracted to the second storage area of the liquid adding assembly, and the second storage area of the liquid adding assembly is inclined to the direction of the incubation disc through the first driving assembly to guide the second antibody reagent into the incubation disc; S6, the incubation disc is incubated with the second antibody; S7, the incubation membrane is taken out, the third extracting cleaning solution is used to clean the incubation disc through the liquid adding avoidance hole, then the incubation disc is inclined through the second driving mechanism to guide the cleaning solution into the waste liquid recovery groove for recovery.

[0032] Those skilled in the art can easily understand that the above is only a preferred embodiment of the present application, and is not used to limit the present application, and any modification, equivalent replacement and improvement within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. A high-throughput protein immunoblotting incubation device, comprising an incubation tray (1), characterized in that: the incubation tray (1) is connected by a support assembly (4) for containing an incubation membrane to be incubated, the support assembly (4) has an upper bearing part (401), a middle bearing part (402) and a lower bearing part (403) arranged in sequence from top to bottom, and the incubation tray (1) is arranged in the middle bearing part (402); a liquid adding assembly (2) is arranged in the upper bearing part (401) of the support assembly (4) by a first driving assembly (5), for driving the liquid adding assembly (2) to add a certain amount of incubation liquid into the incubation tray (1) by the first driving assembly (5); a recovery assembly (3) is arranged in the lower bearing part (403) of the support assembly (4), for recovering the liquid in the incubation tray (1); wherein the liquid adding assembly (2) has at least a first storage area and a second storage area, the first storage area and the second storage area are oppositely distributed, and are used for storing different kinds of incubation liquid to add to the incubation tray (1).

2. The high-throughput protein immunoblotting incubation device according to claim 1, characterized in that: the first storage area (201) and the second storage area (202) of the liquid adding assembly (2) are correspondingly provided with a first guide part (20101) and a second guide part (20201) on the side away from each other, the first guide part (20101) or the second guide part (20201) is used to guide the incubation liquid to flow naturally along the respective guide direction; the incubation tray (1) is open at the upper part, and the volume of the upper opening is greater than the area of the liquid adding assembly (2); wherein the first driving assembly (5) is used to drive the liquid adding assembly (2) to tilt towards the first storage area (201) or the second storage area (202), when the first driving assembly drives the liquid adding assembly (2) to tilt towards the first storage area (201), the incubation liquid in the first storage area (201) flows naturally along the guide direction of the first guide part (20101) to the incubation tray (1), when the first driving assembly drives the liquid adding assembly (2) to tilt towards the second storage area (202), the incubation liquid in the first storage area (201) flows naturally along the guide direction of the second guide part (20201) to the incubation tray (1).

3. The high-throughput protein immunoblotting incubation device according to claim 2, characterized in that: the liquid adding assembly (2) comprises a liquid adding groove, the middle part of the liquid adding groove is divided into the first storage area and the second storage area (202) by a partition, and the bottom of the liquid adding groove is further provided with a sliding groove; the upper bearing part (401) is further provided with a bracket (6), and the sliding groove is slidingly arranged in the bracket (6) and can slide along the length direction of the bracket (6).

4. The high-throughput protein immunoblotting incubation device according to claim 3, characterized in that: ​ ​ ​ ​ ​ ​ ​ ​ ​ The middle part of the liquid adding tank is also provided with a liquid adding avoiding hole (203), which is a cylindrical structure formed by surrounding and has an inlet and an outlet and is located between the first storage area and the second storage area (202); The inlet is flush with the slot of the liquid adding tank, and the outlet faces the incubation disc (1).

5. The high-throughput protein immunoblotting incubation device according to any one of claims 1-4, characterized in that: The incubation disc (1) is arranged on the middle bearing part (402) of the support assembly (4) through a second driving assembly (7), and is used to drive the incubation disc (1) to tilt left and right to shake the incubation liquid in the incubation disc (1).

6. The high-throughput protein immunoblotting incubation device according to claim 5, characterized in that: The incubation disc (1) has a third guide part (101) and a fourth guide part (102), and the third guide part (101) and the fourth guide part (102) are oppositely arranged on the incubation disc (1); A cover (103) is arranged on the upper disc opening of the incubation disc (1), and the cover (103) is provided with a first liquid inlet (10301), a second liquid inlet (10302) and a third liquid inlet (10303), the first liquid inlet (10301) and the second liquid inlet (10302) are oppositely arranged, and the first liquid inlet (10301) and the second liquid inlet (10302) are located directly below the first guide part (20101) and the second guide part (20201) of the liquid adding assembly (2), and are used to receive the incubation liquid guided by the first guide part (20101) or the second guide part (20201); the third liquid inlet (10303) faces the outlet of the liquid adding avoiding hole (203).

7. The high-throughput protein immunoblotting incubation device according to claim 1, characterized in that: The recovery assembly (3) includes a first recovery part and a second recovery part, and the first recovery part and the second recovery part are located directly below the third guide part (101) and the fourth guide part (102); The first recovery part includes a waste liquid recovery tank (301), and the second recovery part includes an incubation liquid recovery pipe (203).

8. The high-throughput protein immunoblotting incubation device according to claim 7, characterized in that: The first guide part (20101), the second guide part (20201), the third guide part (101) and the fourth guide part (102) are all conical.

9. The high-throughput protein immunoblotting incubation device according to claim 8, characterized in that: An upper liquid component (8) is arranged on the support component (4) and above the liquid adding component (2) to supplement the incubation liquid to the first storage area and the second storage area of the liquid adding component (2); the upper liquid component (8) comprises a first upper liquid support (801), a second upper liquid support (802) and a turnover component (803), the first upper liquid support (801) and the second upper liquid support (802) are arranged vertically, and the turnover component (803) is used to drive the first upper liquid support (801) and the second upper liquid support (802) to turn over left and right. The centrifugal tube (9) for filling the incubation liquid is fixedly arranged on the first upper liquid support (801) and the second upper liquid support (802), and when any one of the first upper liquid support (801) or the second upper liquid support (802) is turned over to any one side of left and right, the incubation liquid in the centrifugal tube (9) on the corresponding side is poured into the first storage area or the second storage area, and the centrifugal tube (9) on the first upper liquid support (801) or the second upper liquid support (802) on the side other than the left and right side keeps the tube body and the tube opening upward.

10. A high-throughput protein immunoblotting incubation method characterized in that, The method is applied to the high-throughput protein immunoblotting incubation device in any one of the above claims 1-9, and the method comprises: Step S1, the first time, the cleaning liquid is extracted to clean the incubation disc through the liquid adding avoidance hole, and then the incubation disc is inclined by the second driving mechanism to guide the cleaning liquid into the waste liquid recovery groove for recovery; Step S2, the incubation membrane is placed, the first antibody reagent is extracted to the first storage area of the liquid adding component, and the first storage area of the liquid adding component is driven to incline to the incubation disc direction by the first driving assembly, so that the first antibody reagent is guided into the incubation disc; Step S3, the incubation disc is incubated with the first antibody; Step S4, the second time, the cleaning liquid is extracted to clean the incubation disc through the liquid adding avoidance hole, and then the incubation disc is inclined by the second driving mechanism to guide the cleaning liquid into the waste liquid recovery groove for recovery; Step S5, the second antibody reagent is extracted to the second storage area of the liquid adding component, and the second storage area of the liquid adding component is driven to incline to the incubation disc direction by the first driving assembly, so that the second antibody reagent is guided into the incubation disc; Step S6, the incubation disc is incubated with the second antibody; Step S7, the incubation membrane is taken out, the cleaning liquid is extracted for the third time to clean the incubation disc through the liquid adding avoidance hole, and then the incubation disc is inclined by the second driving mechanism to guide the cleaning liquid into the waste liquid recovery groove for recovery.

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