PVDF (Polyvinylidene Fluoride) membrane incubation device

Abstract

The utility model discloses a PVDF (Polyvinylidene Fluoride) membrane incubation device which comprises a centrifugal machine, a rotating shaft is arranged at the top of the centrifugal machine, the upper end of the rotating shaft is coaxially and fixedly connected with a washing liquid containing barrel, and an annular incubation barrel with an upper opening is arranged around the outer side of the washing liquid containing barrel; the interior of the annular incubation barrel is evenly divided into a plurality of incubation bins through partition plates, a sealing cover is arranged at the top of the annular incubation barrel, a liquid injection opening is formed in the sealing cover, and a liquid injection pump communicated with the liquid injection opening is arranged at the top of the washing liquid containing barrel; a diaphragm fixing frame is vertically inserted into the annular incubation barrel; and a sample feeding opening and a sample discharging opening which are correspondingly communicated with each incubation bin are formed in the outer wall of the annular incubation barrel. The PVDF membrane incubation device has the advantages that the PVDF membrane incubation, washing and waste liquid recovery operation can be carried out in batches, different types of antibodies can be classified, the incubation efficiency is improved, the incubation operation steps are optimized, the incubation effect is ensured, the antibody resources are saved, the antibody waste is reduced, and the PVDF membrane incubation cost input is reduced.

Description

Technical Field

[0001] The utility model relates to the technical field of protein blotting experiments, in particular to a PVDF membrane incubation device. Background Technique

[0002] PVDF membrane (i.e., polyvinylidene fluoride membrane) is a commonly used solid-phase support in protein blotting, with high mechanical strength and hydrophobicity; the pore sizes of PVDF membranes vary, and as the pore size of the membrane continuously decreases, the binding of low-molecular-weight proteins becomes stronger. When in use, it needs to be pretreated with methanol, and the purpose is to activate the positively charged groups on the membrane, making it easier to bind to negatively charged proteins.

[0003] When conducting protein blotting experiments: First, protein samples are electrophoresed in SDS polyacrylamide gel to separate proteins with different molecular weights; then, the proteins on the SDS polyacrylamide gel are transferred to the PVDF membrane by electrophoresis, and the PVDF membrane is cut according to the molecular weight of the protein; after that, the cut corresponding membrane pieces are placed in a primary antibody dilution solution containing the corresponding protein and incubated overnight. After the incubation is completed, the primary antibody dilution solution is recovered, and the membrane is washed with PBS solution at least 3 times, with each washing time lasting up to 3 - 5 minutes; finally, it is incubated with a secondary antibody dilution solution that can specifically bind to the primary antibody at room temperature for about 1 hour. After the incubation is completed, the membrane needs to be washed with PBS solution again 3 times. After washing, a luminescent solution is applied and then color development scanning can be performed on a film scanner.

[0004] Currently, there is no dedicated incubation device for PVDF membranes on the market. Since the incubation solution and washing solution need to be repeatedly changed during the PVDF membrane incubation process, the entire PVDF membrane incubation process is very cumbersome; and because both the primary antibody liquid and the secondary antibody liquid are expensive, usually the price per milliliter is about 20,000 - 100,000. Although it is diluted 1000 - 10000 times during use, due to the large amount and frequency of use, it is still a significant expense. Summary of the Invention

[0005] The purpose of the utility model is to provide a PVDF membrane incubation device aiming at the defects of the prior art.

[0006] To achieve the above purpose, the utility model can adopt the following technical solutions:

[0007] The PVDF membrane incubation device described in the present utility model includes a centrifuge, which has a vertically upward rotating shaft at its top. The upper end of the rotating shaft is coaxially fixedly connected with a washing liquid storage barrel, and an annular incubation barrel with an upper opening is arranged around the outside of the washing liquid storage barrel; the annular incubation barrel is evenly divided into a plurality of incubation chambers by partitions, and the top of the annular incubation barrel has a sealing cover that fits and covers the incubation chambers. The sealing cover has a liquid injection port, and the top of the washing liquid storage barrel has a liquid injection pump connected to the liquid injection port; a membrane fixing frame for clamping the PVDF membrane is vertically inserted in the annular incubation barrel, and a sample addition port and a sample discharge port corresponding to each incubation chamber are arranged on the outer wall of the annular incubation barrel.

[0008] Further, for convenient operation, a liquid crystal control panel is arranged on the side wall of the centrifuge.

[0009] Further, the washing liquid storage barrel is a fully enclosed cylindrical barrel body, and a liquid replenishment port is arranged on the top wall of the washing liquid storage barrel; at this time, a slot is axially opened at the center of the top wall of the washing liquid storage barrel, and a cooling barrel is adaptively inserted in the slot. The barrel wall of the cooling barrel is in contact with the inner wall of the slot, and the washing liquid in the washing liquid storage barrel can be easily cooled through heat conduction.

[0010] Further, for convenient observation, the annular incubation barrel is a transparent barrel body; at this time, scale marks can also be arranged on the outer wall of the annular incubation barrel along the height direction; in addition, the sample addition port is located near the top of the side wall of the annular incubation barrel, and the sample discharge port is located on the bottom wall of the annular incubation barrel.

[0011] Further, a recovery tank is arranged on the top wall of the centrifuge directly below the annular incubation barrel. The recovery tank is an upper-opening annular cavity wound around the outside of the rotating shaft, and a recovery port is arranged on the outer wall of the recovery tank. The sample liquid flowing out from the sample discharge port is collected by the recovery tank and discharged through the recovery port, so as to facilitate the recycling of the sample liquid.

[0012] Further, the membrane fixing frame includes a rectangular frame, the width of the rectangular frame is the same as the width of the incubation chamber, sliding grooves are arranged on the inner sides of the two long sides of the rectangular frame, and an upper clamping plate and a lower clamping plate for clamping the PVDF membrane are slidably arranged in the sliding grooves. The rectangular frame can be stably inserted into the incubation chamber, and the PVDF membrane can be stably clamped by the upper clamping plate and the lower clamping plate thereon, ensuring that PVDF membranes of different lengths can be longitudinally placed in the incubation chamber in an unfolded state.

[0013] The advantages of the present utility model are that it can batch incubate, wash, and recycle waste liquid of PVDF membranes, and can use multiple incubation chambers separated by partition plates to classify different types of antibodies, improve the incubation efficiency, optimize the incubation operation steps, ensure that the PVDF membrane can be fully unfolded during antibody incubation, and at the same time, cooperate with the rotation and shaking of the centrifuge to ensure that the antibody can better bind to the PVDF membrane, ensure the incubation effect, save antibody resources, reduce antibody waste, and reduce the cost investment of PVDF membrane incubation. Brief Description of the Drawings

[0014] Figure 1 is a schematic structural diagram of the present utility model. Detailed Embodiments

[0015] Next, the technical solutions in the embodiments of the present utility model will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present utility model. Obviously, the described embodiments are only a part of the embodiments of the present utility model, rather than all the embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present utility model.

[0016] It should be noted that all directional indications (such as up, down, left, right, front, back...) in the embodiments of the present utility model are only used to explain the relative position relationship and movement conditions between components in a specific posture (as shown in the drawings). If the specific posture changes, the directional indications will also change accordingly.

[0017] As Figure 1 shown, the PVDF membrane incubation device of the present utility model includes a centrifuge 1. A vertically upward rotating shaft 2 is provided at the center of the top of the centrifuge 1, and a convenient-to-operate liquid crystal control panel 3 is provided on the side wall of the centrifuge 1.

[0018] A washing liquid storage bucket 4 is coaxially and fixedly connected to the upper end of the rotating shaft 2. The washing liquid storage bucket 4 is preferably a fully enclosed cylindrical metal bucket body. At this time, a liquid supplement port 5 is provided on the top wall of the washing liquid storage bucket 4 to facilitate supplementing the washing liquid into the washing liquid storage bucket 4 as needed. In addition, in order to facilitate cooling the washing liquid in the washing liquid storage bucket 4, a slot 6 can also be axially opened at the center of the top wall of the washing liquid storage bucket 4. A cooling bucket 7 is adaptively inserted into the slot 6. The cooling bucket 7 is a metal storage bucket with a lid at the upper end, and a cooling liquid can be contained therein. The diameter of the cooling bucket 7 is the same as the diameter of the slot 6, so that when it is inserted into the slot 6, the barrel wall can closely adhere to the inner wall of the slot 6. And by using a metal material with a relatively high thermal conductivity as the washing liquid storage bucket 4 and the cooling bucket 7, the washing liquid in the washing liquid storage bucket 4 can be quickly cooled through heat conduction.

[0019] The outer periphery of the washing liquid storage barrel 4 is provided with an annular incubation barrel 8 with an upper opening, and the annular incubation barrel 8 is fixedly connected to the washing liquid storage barrel 4; the annular incubation barrel 8 is a transparent barrel body, which is convenient to observe the situation inside the barrel at any time. In the annular cavity of the annular incubation barrel 8, a plurality of incubation chambers 10 are separated by a plurality of partition plates 9 evenly arranged at intervals along the circumferential direction; at this time, a sealing cover 11 for covering the incubation chamber 10 is provided on the top of the annular incubation barrel 8. There are a plurality of sealing covers 11, which are respectively and correspondingly adapted to cover the openings of each incubation chamber 10. Each sealing cover 11 is provided with a liquid injection port 12. In this way, a liquid injection pump 13 corresponding to the liquid injection port 12 can be arranged on the top wall of the washing liquid storage barrel 4, so as to be able to correspondingly connect the liquid injection pump 13 and the liquid injection port 12 through a hose, so that the liquid injection pump 13 can be opened to inject the flushing liquid in the washing liquid storage barrel 4 into the corresponding incubation chamber 10. Further, in order to conveniently and accurately observe the liquid injection volume in the incubation chamber 10, a scale mark 14 can also be arranged on the outer wall of the annular incubation barrel 8 along the height direction. The scale marks 14 can be a plurality of ones corresponding to the incubation chambers 10 one by one, so that the liquid volume situation in each incubation chamber 10 can be clearly observed.

[0020] A membrane fixing frame for clamping the PVDF membrane is vertically inserted into the annular incubation barrel 8. The membrane fixing frame includes a rectangular frame 15. The width of the rectangular frame 15 is the same as the width of the incubation chamber 10, and its length is less than the height of the incubation chamber 10. Sliding grooves 16 are arranged on the inner sides of the two long sides of the rectangular frame 15, and an upper clamping plate 17 and a lower clamping plate 18 for clamping the PVDF membrane are slidably arranged in the sliding grooves 16; the rectangular frame 15 of the membrane fixing frame can be stably inserted vertically into the incubation chamber 10, and the upper clamping plate 17 and the lower clamping plate 18 thereon are used to stably clamp the PVDF membrane, ensuring that PVDF membranes of different lengths can be longitudinally placed in the incubation chamber 10 in an unfolded state. In addition, a plurality of membrane fixing frames can be inserted side by side in each incubation chamber 10 according to the use needs.

[0021] A plurality of sample adding ports 19 and sample discharging ports 20 are also arranged on the outer wall of the annular incubation barrel 8. Each group of sample adding ports 19 and sample discharging ports 20 is correspondingly communicated with an incubation chamber 10. Among them, the sample adding port 19 is located near the top of the side wall of the annular incubation barrel 8, and the sample discharging port 20 is located on the bottom wall of the annular incubation barrel 8. At this time, a recovery groove 21 can be arranged on the top wall of the centrifuge 1 directly below the annular incubation barrel 8. In order to avoid affecting the rotation of the rotating shaft 2, the recovery groove 21 is an upper-opening annular groove cavity that is intermittently wound around the outside of the rotating shaft 2. A recovery port 22 is also arranged on the outer wall of the recovery groove 21 near the bottom. The sample liquid flowing out from the sample discharging port 20 is collected by the recovery groove 21 and collected through the recovery port 22, so as to facilitate the recycling of the sample liquid.

[0022] During use, first, the cut PVDF membranes are successively clamped on each membrane holder, keeping the PVDF membranes in an unfolded state. Then, according to the experiment, the PVDF membranes that need to be added with the same type of antibody (primary antibody, secondary antibody) are inserted side by side into the same incubation chamber 10, and the corresponding primary antibody dilution is added to the corresponding incubation chamber 10 from the sample addition port 19 according to the classification. The temperature of the added primary antibody dilution can be directly controlled at about 4°C, and the added height should cover the PVDF membrane. After that, start the centrifuge 1 and shake it by rotation to make the primary antibody bind better to the PVDF membrane.

[0023] After the primary antibody incubation is completed, the primary antibody dilution is discharged through the sample discharge port 20. During the drainage process, the centrifuge 1 can also be started to rotate slowly, so as to ensure that the primary antibody dilution is completely drained into the recovery tank 21 below through centrifugation, and then the used primary antibody dilution is recovered through the recovery port 22 on the recovery tank 21. Note that when recovering the primary antibody dilution, the sample discharge ports 20 at the bottom of each incubation chamber 10 should be opened separately according to the specific situation to collect the primary antibody dilution separately.

[0024] After the collection of the primary antibody dilution is completed, the liquid injection pump 13 can be started to inject the washing solution (i.e., PBS solution) into the incubation chamber 10 to wash the PVDF membrane after incubating with the primary antibody. The membrane washing is carried out three times, 3 - 5 minutes for each time. During the membrane washing process, the cooling bucket 7 can be inserted into the slot 6 on the top wall of the washing solution storage bucket 4, so that the temperature of the washing solution is always maintained at about 4°C.

[0025] After the membrane washing is completed, the secondary antibody incubation can be carried out in the same operation mode, as well as the membrane washing again after the secondary antibody incubation is completed. The whole operation process realizes the batch incubation, washing and waste liquid recovery of the PVDF membrane, and can use the multiple incubation chambers 10 separated by the partition 9 to realize the classification incubation of different types of antibodies, improve the incubation efficiency, optimize the incubation operation steps, ensure that the PVDF membrane can be completely unfolded during the antibody incubation process, and at the same time, cooperate with the rotation and shaking of the centrifuge 1 to ensure that the antibody can bind better to the PVDF membrane, ensure the incubation effect, save antibody resources, reduce antibody waste, and reduce the cost input of PVDF membrane incubation.

Claims

1. A PVDF membrane incubation device, characterized in that: It comprises a centrifuge, the top of which is provided with a vertically upward rotating shaft, the upper end of the rotating shaft is coaxially fixedly connected with a washing liquid containing barrel, the outer side of the washing liquid containing barrel is surrounded by an annular incubation barrel with an upper opening; the inside of the annular incubation barrel is evenly divided into a plurality of incubation chambers by partitions, the top of the annular incubation barrel is provided with a sealing cover adapted to cover the incubation chambers, the sealing cover is provided with a liquid injection port, the top of the washing liquid containing barrel is provided with an injection pump connected with the liquid injection port; a membrane fixing frame for clamping a PVDF membrane is vertically inserted in the annular incubation barrel, and a sample addition port and a sample discharge port corresponding to each of the incubation chambers are provided on the outer wall of the annular incubation barrel.

2. The PVDF membrane incubation device according to claim 1, characterized in that: The washing liquid containing barrel is a fully enclosed cylindrical barrel body, and a liquid replenishing port is arranged on the top wall of the washing liquid containing barrel.

3. The PVDF membrane incubation device according to claim 2, characterized in that: A slot is axially provided at the center of the top wall of the washing liquid containing barrel, and a cooling barrel is adapted to be inserted into the slot.

4. The PVDF membrane incubation device according to claim 1, characterized in that: The annular incubation barrel is a transparent barrel body.

5. The PVDF membrane incubation device according to claim 4, characterized in that: Scale markings are arranged on the outer wall of the annular incubation barrel along the height direction.

6. The PVDF membrane incubation device according to claim 4, characterized in that: The sample addition port is located on the side wall of the annular incubation barrel near the top, and the sample discharge port is located on the bottom wall of the annular incubation barrel.

7. The PVDF membrane incubation device according to claim 6, characterized in that: A recovery groove is arranged on the top wall of the centrifuge, facing the bottom of the annular incubation barrel. The recovery groove is an upper opening annular groove cavity arranged around the outer side of the rotating shaft, and a recovery port is arranged on the outer wall of the recovery groove.

8. The PVDF membrane incubation device according to claim 1, characterized in that: The membrane fixing frame comprises a rectangular frame, the width of which is consistent with the width of the incubation chamber, and slide grooves are arranged inside the two long sides of the rectangular frame, and an upper clamping plate and a lower clamping plate for clamping the PVDF membrane are slidably arranged in the slide grooves.

9. The PVDF membrane incubation device according to claim 1, characterized in that: A liquid crystal control panel is arranged on the side wall of the centrifuge.

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