A cell culture plate surface treatment device and treatment process

Abstract

The present invention discloses a cell culture plate surface treatment device and treatment process, which relates to the field of surface treatment technology, including a conveying component, an immersion component and a spray component, and also includes: an injection component and a transport component, the transport component is arranged outside the conveying component, and the transport component is used to take the cell culture plate and shake it; the immersion component includes a transfer plate, the bottom of the transfer plate is provided with a mounting groove, the inside of the mounting groove is connected with a blocking plate by screws, the blocking plate is fitted with the inner wall of the mounting groove, and the blocking plate and the cell culture plate are squeezed and matched to separate each culture hole into a closed chamber. The present invention blocks each culture hole into an independent chamber by setting a blocking plate, and an initiator can be injected into each independent culture hole during immersion, so that chemical immersion only exists in each culture hole that needs to be hydrophilically treated, which not only effectively saves the initiator, but also can avoid the narrow part of the culture plate being affected by the hydrophilic treatment and subsequent use.

Description

Technical Field

[0001] The invention relates to the technical field of surface treatment, and in particular to a surface treatment device and a treatment process for a cell culture plate. Background Art

[0002] Cell culture plates are important tools for cell culture and observation in laboratories. Their main purpose is to provide a suitable growth environment for cells so as to conduct various biological experiments, such as cell proliferation, differentiation, drug screening, etc. Among them, the culture of adherent cells needs to be able to attach to the surface of the culture medium, such as fibroblasts, epithelial cells and many cancer cells. In order to support the growth of such cells, the inner wall of the culture plate needs to be hydrophilic before culture.

[0003] The above-mentioned hydrophilic treatment is a surface modification technology that aims to change the physical and chemical properties of the material surface to make it more hydrophilic. Common methods for hydrophilic treatment include physical treatment and chemical treatment; among them, chemical treatment includes spraying or immersion. The above methods all coat a layer of chemical substances on the surface of the culture plate to improve its hydrophilic properties, thereby enhancing the attachment and growth of cells on the culture plate. Compared with spraying, chemical immersion can achieve modification treatment more evenly and comprehensively.

[0004] However, in actual application, the method of immersing the cell culture plate in an initiator for modification has the problems of initiator waste and over-treatment of the cell culture plate. Since the entire cell culture plate needs to be placed in the initiator for immersion during the modification process, multiple parts such as the outer wall and the bottom of the cell culture plate usually do not need to be hydrophilic treated. The direct immersion method will inevitably cause all parts of the cell culture plate to be hydrophilic treated, thereby causing a waste of initiator. As the number of immersion parts increases, the degree of contamination of the initiator after immersion will also increase, especially the traditional immersion needs to be carried out in a container that is relatively easy to place, which makes the initiator more easily contaminated. As for the cell culture plate itself, since the hydrophilic treated parts are more likely to adhere to water droplets and other microorganisms, the hydrophilic treatment of the parts that do not need hydrophilic treatment will also have a certain impact on the culture process.

[0005] like Fig.10The invention relates to a universal ELISA plate for cell culture, wherein a is a cell culture plate, b is a top groove, c is a culture well, and d is a part that does not need to be hydrophilic. The structure is that each culture well has an independent protruding part, so that the overall structure of the culture plate is a plate with a groove on the top, and there are a number of cylindrical culture wells in the groove. For this type of culture plate, the parts in the groove other than the culture wells do not need to be hydrophilic (as shown in the figure), but based on this arrangement, the parts will be hydrophilic during the chemical immersion process, so that the parts are more likely to adhere to water droplets and pollutants during the actual operation. Since the parts are narrow, it is not easy to clean them. Moreover, due to their hydrophilic characteristics, the drying process after cleaning will be more difficult, which is not conducive to practical application. Summary of the invention

[0006] The purpose of the present invention is to provide a cell culture plate surface treatment device and treatment process to solve the problem that when the cell culture plate is directly immersed in the modified state, the immersion liquid is wasted and polluted, and the cell culture plate is easily contaminated.

[0007] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0008] A cell culture plate surface treatment device comprises: a conveying component, the conveying component is used to convey the cell culture plate, the conveying component comprises a base and a conveyor belt; a stopper is fixedly connected to the inner side of the base; an immersion component, the immersion component is used to chemically immerse the cell culture plate; a spray component, the spray component is used to spray a grafting monomer solution on the cell culture plate after chemical immersion; the immersion component comprises a transfer plate, the bottom of the transfer plate is provided with a mounting groove, the interior of the mounting groove is connected to a blocking plate by screws, the blocking plate is fitted with the inner wall of the mounting groove, the top and bottom of the blocking plate are fixedly connected with sealing strips, the sealing strips are tightly fitted with the mounting groove, and the blocking plate is squeezed and matched with the cell culture plate Each culture well is divided into a closed chamber, and the top of the transfer plate is fixedly connected to a cover shell; a transfer cavity is opened on the top of the sealing plate, and a plurality of liquid outlets are fixedly connected to the bottom of the sealing plate, and a through hole for connecting the liquid outlets with the transfer cavity is opened on the sealing plate; an input pipe connected to the mounting groove is fixedly connected to the top of the transfer plate; a plurality of sealing rings are fixedly connected to the bottom of the sealing plate, and the sealing rings correspond to the liquid outlets one by one; it also includes: a transfer component, which is arranged on the outside of the conveying component, and the transfer component is used to take the cell culture plate and shake it; an injection component, which is arranged above the cover shell, and the injection component is used to provide an initiator to the immersion component through the input pipe.

[0009] By adopting the above technical scheme, each culture well is sealed into an independent chamber by setting a sealing plate, and an initiator can be injected into each independent culture well during immersion, so that chemical immersion only exists in each culture well that needs to be hydrophilic treated, thereby achieving targeted modification treatment. It not only effectively saves the initiator, but also avoids the narrow part of the culture plate being affected by the hydrophilic treatment and affecting subsequent use.

[0010] A further improvement of the technical solution of the present invention is that: the transfer component includes a servo module symmetrically installed on both sides of the base of the conveying component, the movable end of the servo module is fixedly connected to a U-shaped frame, the top of the inner side of the U-shaped frame is fixedly connected to a first cylinder, the piston rod of the first cylinder is fixedly connected to a lifting plate, the top of the lifting plate is fixedly connected to a motor, the output end of the motor extends to the bottom of the lifting plate and is fixedly connected to a rocker, and the cover shell is rotatably connected to the bottom of the rocker; the two sides of the cover shell are symmetrically fixedly connected to extension plates; the bottom of the lifting plate is rotatably connected to an adapter block, the bottom of the adapter block is rotatably connected to an adapter, and the bottom of the adapter is rotatably connected to the extension plate; the top of the transfer plate is fixedly connected to an intake pipe, and a through hole connecting the intake pipe and the non-culture hole part of the cell culture plate is provided on the transfer plate; one side of the base is fixedly connected to an air pump, and the air pump is connected to the intake pipe.

[0011] By adopting the above technical solution, the motor is controlled to drive the rocker to rotate, and further, the cover shell and the structure connected thereto will rotate around the motor output shaft; and the cover shell is limited by the adapter block and the adapter at the same time, so the direction will not change, but the rotation will be completed while the direction remains unchanged, thereby achieving shaking, so that the initiator can move along the inner wall of the culture hole.

[0012] A further improvement of the technical solution of the present invention is that: a plurality of collecting bars are fixedly connected to the top of the transfer plate, a collecting pipe is commonly connected to the tops of the collecting bars, an electric valve is fixedly connected to one side of the transfer plate, one end of the collecting pipe is connected to the electric valve; a whistle is connected to the other end of the electric valve, a plurality of pressure-discharging pipes are fixedly connected to the bottom of the inner wall of the transfer cavity, a plurality of through holes corresponding to the pressure-discharging pipes are opened at the top of the inner wall of the mounting groove, and the through holes are connected to the collecting bars; an air outlet hole connected to the pressure-discharging pipe is opened at the bottom of the sealing plate.

[0013] By adopting the above technical solution, the electric valve is kept turned on, and then the air pump is controlled to work. The air pump draws air into the installation groove through the suction pipe, and the suction position is a non-culture hole position. Since this position is fitted with the sealing plate and a relatively closed chamber is formed, the suction can make the culture plate adsorbed on the sealing plate to complete the adsorption of the culture plate; further, if the sealing of each culture hole is poor, air leakage will occur. At this time, suction of this position will continuously draw air from the culture hole, so that the initiator in the transfer cavity is drawn into the unsealed culture hole.

[0014] A further improvement of the technical solution of the present invention is that: a horizontal plate is fixedly connected between the inner walls of the air outlet, a sliding rod is slidably connected to the bottom of the horizontal plate, a plug is fixedly connected to the bottom of the sliding rod, a floating ball is fixedly connected to the bottom of the plug, a sliding pin is fixedly connected to the outside of the sliding rod, the top of the sliding rod extends to the top of the horizontal plate and is fixedly connected to a limiting plate; a plurality of strip grooves are provided on the outside of the sliding pin.

[0015] By adopting the above technical solution, with the continuous injection of initiator, the liquid level in the culture well rises, thereby causing the float to float up and driving the plug and the sliding pin to move upward. When the culture well is filled with initiator, the plug is completely blocked, so the corresponding culture well can no longer be injected with initiator. At the same time, since the plug blocks the corresponding air outlet, the initiator will not overflow. When all culture wells are filled, there is obviously a large air pressure resistance when the piston plate continues to press downward.

[0016] A further improvement of the technical solution of the present invention is that the injection component includes a mounting frame, the mounting frame is fixedly connected to the top of the lifting plate, the top of the mounting frame is fixedly connected to the second cylinder, the inner side of the mounting frame is fixedly connected to the piston cylinder, the inner walls of the piston cylinder are slidably connected with a piston plate, the piston rod of the second cylinder extends to the interior of the piston cylinder and is fixedly connected to the piston plate; a pressure regulating tube is provided on the piston cylinder, a valve is provided on the pressure regulating tube, and the bottom of the piston cylinder is connected to the input pipe.

[0017] By adopting the above technical solution, the piston rod of the second cylinder is controlled to extend and the piston plate is pushed downward, thereby squeezing the initiator in the transfer chamber into each culture hole; further, after the immersion is completed, the second cylinder is controlled to retract and the piston plate is driven to move upward, thereby applying negative pressure to the piston cylinder, so that the initiator in the culture hole is sucked into the transfer chamber and can be used multiple times. During this process, external air enters the transfer chamber from the air outlet and pushes the plug downward.

[0018] A further improvement of the technical solution of the present invention is that a force-bearing plate is fixedly connected to the inner side of the base, and the top of the conveyor belt is higher than the top of the force-bearing plate.

[0019] By adopting the above technical solution, the top of the conveyor belt is initially higher than the force plate, so the conveyor belt provides support and transport for the culture plate. When pressed down, the force plate is flush with the conveyor belt. At this time, the culture plate is blocked by the block and cannot move on the one hand, and supported by the force plate on the other hand.

[0020] A further improvement of the technical solution of the present invention is that the interior of the liquid outlet nozzle is arranged as a honeycomb hole structure.

[0021] By adopting the above technical solution, by setting the liquid outlet nozzle as a honeycomb hole structure, the channel flowing downward from the transfer chamber is divided into multiple slender channels. It is difficult for the initiator to flow out if the pressure inside the transfer chamber is not released, thereby avoiding the dripping of the initiator.

[0022] A further improvement of the technical solution of the present invention is that: an annular plate is fixedly connected to the bottom of the lifting plate, and the bottom of the annular plate and the top of the rocker are both set as rough surfaces and contact each other.

[0023] By adopting the above technical solution, the rocker will rub against the annular plate during rotation, and the contact surface of the two is a rough surface, so high-frequency jitter will be generated during relative friction, thereby making the initiator concentration distribution more uniform and easier to diffuse to various parts.

[0024] The present invention also provides a cell culture plate surface treatment process, comprising the following steps:

[0025] S1. Soak the cell culture plate in anhydrous ethanol and then rinse it repeatedly with deionized water;

[0026] During the production and storage process, cell culture plates will accumulate dust, grease, dirt or other contaminants. By cleaning, these impurities can be effectively removed, providing a clean base for chemical modification.

[0027] S2. Completely immerse the cleaned cell culture plate in a sulfuric acid-nitric acid mixed acid solution at 25-40° C. for 4-6 hours; take it out and repeatedly wash it with deionized water, and dry it;

[0028] S3, blocking the top of the cell culture plate so that each culture well is separated into a relatively closed chamber;

[0029] Since there are quite a few parts on the cell culture plate that do not need to be hydrophilic treated, especially some narrow areas, which are very easy to adhere to water droplets after hydrophilic treatment, thus providing an attachment environment for microorganisms and impurities and are not convenient for cleaning, they are separated and treated in a targeted manner.

[0030] S4, injecting the initiator into each culture well until the culture well is completely filled, and leaving it to soak for 24 hours, shaking the cell culture plate for 10 seconds every hour at a position deviated from the center of the cell culture plate to make the initiator concentration evenly distributed, and extracting the initiator after the soaking is completed;

[0031] During the static immersion process, the initiator will gradually settle, making the concentration at the bottom higher than that at the top, which will result in differences in treatment effects. Therefore, the cell culture plate is shaken between treatments to improve the uniformity of treatment.

[0032] S5, spraying the grafting monomer solution onto the cell culture plate and letting it stand for 1 hour. After the reaction is completed, repeatedly rinsing with ethanol solution and deionized water to remove the unreacted initiator and grafting monomer, and drying;

[0033] By reacting the grafting solution with the initiator, a hydrophilic layer can be produced on the inner wall of the culture well of the cell culture plate, while other parts will not be hydrophilized.

[0034] Due to the adoption of the above technical solution, the present invention has the following technical advances compared with the prior art:

[0035] 1. The present invention seals each culture well into an independent chamber by setting a sealing plate. During immersion, an initiator can be injected into each independent culture well, so that chemical immersion only exists in each culture well that needs to be hydrophilic treated, thereby achieving targeted modification treatment. It not only effectively saves the initiator, but also avoids the narrow part of the culture plate being affected by the hydrophilic treatment and affecting subsequent use.

[0036] 2. The present invention provides a structure capable of shaking the culture plate, specifically by controlling the motor to drive the rocker to rotate, so that the cover shell and the structure connected thereto rotate around the motor output shaft; and the cover shell is limited by the adapter block and the adapter at the same time, so the direction will not change, but the rotation is completed without changing the direction, thereby achieving shaking, so that the initiator can move along the inner wall of the culture hole.

[0037] 3. The present invention provides a suction cup structure capable of detecting air tightness. When in use, the air pump is controlled to make the suction pipe suck air into the installation groove, and the suction position is a non-culture hole position. Since this position is fitted with the sealing plate to form a relatively closed chamber, the suction can make the culture plate adsorbed on the sealing plate to complete the adsorption of the culture plate. If the sealing of each culture hole is poor, air leakage will occur. At this time, suction of this position will continuously suck air from the culture hole, so that the initiator in the transfer cavity is drawn into the unsealed culture hole, which will cause a whistle to sound, so as to realize the detection of the sealing before immersion. If a sealing problem occurs in the above process, the initiator will enter the corresponding culture hole, so that the leakage position can be directly judged, which greatly facilitates the practical application.

[0038] 4. The present invention provides a float structure. With the continuous injection of the initiator, the liquid level in the culture well rises, so that the float floats up and drives the plug and the sliding pin to move upward. When the culture well is filled with the initiator, the plug is completely blocked, so the corresponding culture well can no longer be injected with initiator. At the same time, since the plug blocks the corresponding air outlet, the initiator will not overflow. When all the culture wells are filled, the piston plate continues to press down and there is obviously a large air pressure resistance, which can prevent the continued injection of the initiator. Furthermore, the increase in injection pressure will be fed back to the controller, so that the injection of the initiator can be stopped.

[0039] 5. The present invention arranges a suction structure to squeeze in or extract the initiator. During operation, the piston rod of the second cylinder is controlled to extend to push the piston plate downward, thereby squeezing the initiator in the transfer chamber into each culture hole. After the soaking is completed, the second cylinder is controlled to retract to drive the piston plate upward, thereby applying negative pressure to the piston cylinder, so that the initiator in the culture hole is sucked into the transfer chamber and can be used multiple times. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] The present invention will be further described below in conjunction with the accompanying drawings.

[0041] Figure 1 It is a schematic diagram of the structure of the present invention from the first viewing angle as a whole;

[0042] Figure 2 It is a schematic diagram of the structure of the second viewing angle of the whole invention;

[0043] Figure 3 A schematic diagram of the U-shaped frame and internal structure of the present invention;

[0044] Figure 4 It is a schematic cross-sectional structure diagram of the soaking component of the present invention;

[0045] Figure 5 It is a schematic cross-sectional structural diagram of the injection component of the present invention;

[0046] Figure 6 It is a schematic diagram of the external structure of the immersion component of the present invention;

[0047] Figure 7 It is a schematic structural diagram of a cross-section of a plugging plate of the present invention;

[0048] Figure 8 It is a schematic diagram of the disassembly of the sliding pin, the plug and the floating ball of the present invention;

[0049] Fig. 9 It is a schematic cross-sectional structural diagram of the liquid outlet nozzle of the present invention;

[0050] Fig.10 It is a schematic diagram of the structure of an existing universal ELISA plate cell culture plate;

[0051] Fig.11 For the present invention Figure 1 Enlarged view of point A in the middle;

[0052] Fig.12 For the present invention Figure 2 Enlarged view of point B in the middle;

[0053] Fig.13 For the present invention Figure 3 Enlarged view of point C in the middle;

[0054] Fig.14 For the present invention Figure 7 Enlarged view of point D in the middle.

[0055] In the figure: 1. conveying component; 2. spray component; 3. servo module; 4. U-shaped frame; 5. first cylinder; 6. lifting plate; 7. motor; 8. cover; 9. transfer plate; 10. blocking plate; 11. extension plate; 12. liquid outlet; 13. sealing ring; 14. electric valve; 15. whistle; 16. air outlet; 17. horizontal plate; 18. sliding pin; 19. plug; 20. floating ball; 21. sliding rod; 2 2. Limit plate; 23. Honeycomb hole structure; 24. Mounting frame; 25. Second cylinder; 26. Piston barrel; 27. Piston plate; 28. Adapter block; 29. ​​Adapter; 30. Pressure regulating pipe; 31. Ring plate; 32. Rocker; 33. Force plate; 34. Stopper; 35. Manifold; 36. Collecting bar; 37. Transfer chamber; 38. Mounting groove; 39. Input pipe; 40. Intake pipe; 41. Exhaust pipe. DETAILED DESCRIPTION

[0056] The present invention is further described in detail below in conjunction with embodiments:

[0057] Example 1

[0058] like Figure 1 , Figure 4 and Figure 7As shown, the present invention provides a cell culture plate surface treatment device, comprising: a conveying component 1, the conveying component 1 is used to convey the cell culture plate, and the conveying component 1 includes a base and a conveyor belt; a block 34 is fixedly connected to the inner side of the base; an immersion component, the immersion component is used to chemically immerse the cell culture plate; a spray component 2, the spray component 2 is used to spray a grafting monomer solution on the cell culture plate after chemical immersion; the immersion component includes a transfer plate 9, the bottom of the transfer plate 9 is provided with a mounting groove 38, the interior of the mounting groove 38 is connected to a sealing plate 10 by screws, the sealing plate 10 is fitted with the inner wall of the mounting groove 38, the top and bottom of the sealing plate 10 are fixedly connected with sealing strips, the sealing strip on the top of the sealing plate 10 is tightly fitted with the mounting groove 38, and the sealing plate 10 and the cell culture plate The extrusion fit separates each culture hole into a closed chamber, and the top of the transfer plate 9 is fixedly connected to the cover shell 8; the top of the sealing plate 10 is provided with a transfer chamber 37, and the bottom of the sealing plate 10 is fixedly connected with a plurality of liquid outlets 12, and the sealing plate 10 is provided with a through hole for connecting the liquid outlets 12 with the transfer chamber 37; the top of the transfer plate 9 is fixedly connected with an input pipe 39 connected with the mounting groove 38; the bottom of the sealing plate 10 is fixedly connected with a plurality of sealing rings 13, and the sealing rings 13 correspond to the liquid outlets 12 one by one; it also includes: a transfer component, which is arranged on the outside of the conveying component 1, and the transfer component is used to take the cell culture plate and shake it; an injection component, which is arranged above the cover shell 8, and the injection component is used to provide an initiator to the immersion component through the input pipe 39.

[0059] In this embodiment, each culture well is sealed into an independent chamber by setting a sealing plate 10, and an initiator can be injected into each independent culture well during immersion, so that chemical immersion only exists in each culture well that needs to be hydrophilic treated, thereby achieving targeted modification treatment, which not only effectively saves the initiator, but also prevents the narrow part of the culture plate from being affected by the hydrophilic treatment and affecting subsequent use.

[0060] Wherein, the above-mentioned chemical immersion is an immersion initiator.

[0061] like Figure 3 , Figure 7 and Fig.13As shown, preferably, the transfer component includes a servo module 3 symmetrically installed on both sides of the base of the conveying component 1, the movable end of the servo module 3 is fixedly connected to a U-shaped frame 4, the top of the inner side of the U-shaped frame 4 is fixedly connected to a first cylinder 5, the piston rod of the first cylinder 5 is fixedly connected to a lifting plate 6, the top of the lifting plate 6 is fixedly connected to a motor 7, the output end of the motor 7 extends to the bottom of the lifting plate 6 and is fixedly connected to a rocker 32, and the cover 8 is rotatably connected to the bottom of the rocker 32; the two sides of the cover 8 are symmetrically fixedly connected to the extension plate 11; the bottom of the lifting plate 6 is rotatably connected to a transfer block 28, the bottom of the transfer block 28 is rotatably connected to a transfer piece 29, and the bottom of the transfer piece 29 is rotatably connected to the extension plate 11; the top of the transfer plate 9 is fixedly connected to an air suction pipe 40, and a through hole connecting the air suction pipe 40 and the non-culture hole part of the cell culture plate is opened on the transfer plate 9; one side of the base is fixedly connected to an air pump, and the air pump is connected to the air suction pipe 40.

[0062] The coordination between the plugging time of the plug 19 and the liquid level inside the culture well is usually achieved by controlling the properties (density and volume) of the float 20, but there may be errors in the actual operation (that is, the above-mentioned precise control is usually only achieved under ideal conditions, and errors need to be considered in the actual operation). Therefore, the float 20 usually pushes the plug 19 to completely block the vent 16 under the action of buoyancy before the culture well is filled, and the remaining small part that has not been immersed needs to be compensated by shaking the culture plate;

[0063] In this embodiment, by controlling the operation of the motor 7, the rocker 32 is driven to rotate, and further, the cover 8 and the structure connected thereto are caused to rotate around the output shaft of the motor 7; and the cover 8 is limited by the adapter block 28 and the adapter 29 at the same time, so the direction will not change, but the rotation is completed while the direction remains unchanged, thereby achieving shaking, so that the initiator can move along the inner wall of the culture hole.

[0064] Preferably, the adapter block 28 and the adapter 29 are hollow structures that are connected to each other internally, and the rotational connection portion between the adapter block 28 and the housing is a rotary joint, and a rotary joint is also provided on the top of the adapter block 28. The air supply of the air pump and the injection of the initiator are respectively connected to the adapter block 28 through the rotary joint, and further connected to the bottom of the housing 8, so that the pipeline will not be entangled during the flipping process. The specific structure is as follows Figure 5 shown.

[0065] Example 2

[0066] like Figure 4 , Figure 7 and Fig.14As shown, on the basis of Example 1, the present invention provides a technical solution: preferably, a plurality of collecting bars 36 are fixedly connected to the top of the transfer plate 9, and a collecting pipe 35 is commonly connected to the top of the collecting bars 36, an electric valve 14 is fixedly connected to one side of the transfer plate 9, and one end of the collecting pipe 35 is connected to the electric valve 14; a whistle 15 is connected to the other end of the electric valve 14, a plurality of pressure-discharging pipes 41 are fixedly connected to the bottom of the inner wall of the transfer chamber 37, a plurality of through holes corresponding to the pressure-discharging pipes 41 are opened at the top of the inner wall of the mounting groove 38, and the through holes are connected to the collecting bars 36; an air outlet 16 connected to the pressure-discharging pipe 41 is opened at the bottom of the sealing plate 10.

[0067] In this embodiment, the electric valve 14 is kept turned on, and the air pump is controlled to work. The air pump sucks air into the mounting groove 38 through the air suction pipe 40, and the air suction part is the non-culture hole part. Since this part is attached to the blocking plate 10 and a relatively closed chamber is formed, the air suction can make the culture plate adsorbed on the blocking plate 10, thereby completing the adsorption of the culture plate;

[0068] Furthermore, if the sealing of each culture hole is poor, air leakage will occur. At this time, suction at this part will continuously suck air from the culture hole, so that the initiator in the transfer chamber 37 is drawn into the unsealed culture hole.

[0069] Example 3

[0070] like Figure 4 , Figure 7 and Figure 8 As shown, on the basis of Example 1, the present invention provides a technical solution: preferably, a transverse plate 17 is fixedly connected between the inner walls of the air outlet 16, a slide rod 21 is slidably connected to the bottom of the transverse plate 17, a plug 19 is fixedly connected to the bottom of the slide rod 21, a float 20 is fixedly connected to the bottom of the plug 19, a slide pin 18 is fixedly connected to the outer side of the slide rod 21, the top of the slide rod 21 extends to the top of the transverse plate 17 and is fixedly connected to the limiting plate 22; a plurality of strip grooves are provided on the outer side of the slide pin 18.

[0071] When injecting the initiator into the culture well, the operator cannot intuitively judge the injection amount of the initiator, and because each culture well is in an independent liquid level state, the amount of initiator in each culture well cannot be reflected by controlling the total amount injected, so the injection amount of the initiator cannot be accurately controlled;

[0072] In this embodiment, with the continuous injection of the initiator, the liquid level in the culture well rises, so that the float 20 floats up and drives the plug 19 and the sliding pin 18 to move upward. When the culture well is filled with the initiator, the plug 19 is completely blocked, so the initiator can no longer be injected into the corresponding culture well. At the same time, since the plug 19 blocks the corresponding air outlet 16, the initiator will not overflow. When all the culture wells are filled, there is obviously a large air pressure resistance when the piston plate 27 continues to press down.

[0073] Example 4

[0074] like Figure 3 and Figure 5 As shown, on the basis of Example 1, the present invention provides a technical solution: preferably, the injection component includes a mounting frame 24, the mounting frame 24 is fixedly connected to the top of the lifting plate 6, the top of the mounting frame 24 is fixedly connected to the second cylinder 25, the inner side of the mounting frame 24 is fixedly connected to the piston cylinder 26, the inner wall of the piston cylinder 26 is slidably connected with a piston plate 27, the piston rod of the second cylinder 25 extends to the inside of the piston cylinder 26 and is fixedly connected to the piston plate 27; a pressure regulating pipe 30 is provided on the piston cylinder 26, a valve is provided on the pressure regulating pipe 30, and the bottom of the piston cylinder 26 is connected to the input pipe 39.

[0075] In this embodiment, the piston rod of the second cylinder 25 is controlled to extend to push the piston plate 27 downward, thereby squeezing the initiator in the transfer chamber 37 into each culture well;

[0076] Furthermore, after the soaking is completed, the second cylinder 25 is controlled to be retracted, driving the piston plate 27 to move upward, thereby drawing negative pressure on the piston cylinder 26, so that the initiator in the culture hole is sucked into the transfer chamber 37, which can be used multiple times. During this process, the external air enters the transfer chamber 37 from the air outlet 16 and pushes the plug 19 downward. When the liquid level drops to a certain level, the plug 19 can also be separated from the air outlet 16 under the action of its own gravity;

[0077] There is a small gap between the bottom of the liquid outlet nozzle 12 and the bottom of the culture hole.

[0078] For the pressure regulating tube 30, when the culture hole is not filled during operation, the second cylinder 25 is pushed out to the longest state. At this time, if space is needed to continue pressurizing, the valve can be opened, and then the second cylinder 25 can be controlled to be retracted, and then the valve can be closed, so that the second cylinder 25 can be used to provide squeezing force again (it is not needed under normal circumstances, and it is intended to temporarily solve the situation where a malfunction occurs during operation).

[0079] like Figure 1 , Figure 2 , Fig.11 and Fig.12As shown, preferably, a force-bearing plate 33 is fixedly connected to the inner side of the base, and the top of the conveyor belt is higher than the top of the force-bearing plate 33 .

[0080] When the blocking plate 10 is pressed down, the conveyor belt will sink downward and cannot provide sufficient rigid support;

[0081] In this embodiment, the top of the conveyor belt is initially higher than the force plate 33, so the conveyor belt provides support and transport for the culture plate. When pressed down, the force plate 33 is flush with the conveyor belt. At this time, the culture plate is blocked by the block 34 and cannot move on the one hand, and is supported by the force plate 33 on the other hand.

[0082] like Figure 4 , Figure 6 and Fig. 9 As shown, preferably, the interior of the liquid outlet nozzle 12 is configured as a honeycomb hole structure 23 .

[0083] Since the amount of initiator in the transfer chamber 37 is completely controlled by the air pressure in the piston cylinder 26, and the liquid outlet 12 for extruding the initiator is in a conducting state during the whole process, the initiator is likely to leak out during the operation, especially when the servo module 3 is working or the motor 7 is working;

[0084] In this embodiment, by setting the liquid outlet nozzle 12 as a honeycomb hole structure 23, the channel flowing downward from the transfer chamber 37 is divided into multiple slender channels. It is difficult for the initiator to flow out if the internal pressure of the transfer chamber 37 is not released, thereby avoiding the dripping of the initiator.

[0085] like Figure 3 and Fig.13 As shown, preferably, the bottom of the lifting plate 6 is fixedly connected with an annular plate 31, and the bottom of the annular plate 31 and the top of the rocker 32 are both set as rough surfaces and contact each other.

[0086] In this embodiment, the rocker 32 will rub against the annular plate 31 during its rotation, and the contact surfaces of the two are rough surfaces, so high-frequency jitter will be generated during the relative friction, thereby making the initiator concentration distribution more uniform and easier to diffuse to various parts.

[0087] The present invention also provides a cell culture plate surface treatment process, comprising the following steps:

[0088] S1. Soak the cell culture plate in anhydrous ethanol and then rinse it repeatedly with deionized water;

[0089] During the production and storage process, cell culture plates will accumulate dust, grease, dirt or other contaminants. By cleaning, these impurities can be effectively removed, providing a clean base for chemical modification.

[0090] S2. Completely immerse the cleaned cell culture plate in a sulfuric acid-nitric acid mixed acid solution at 25-40° C. for 4-6 hours; take it out and repeatedly wash it with deionized water, and dry it;

[0091] S3, blocking the top of the cell culture plate so that each culture well is separated into a relatively closed chamber;

[0092] Since there are quite a few parts on the cell culture plate that do not need to be hydrophilic treated, especially some narrow areas, which are very easy to adhere to water droplets after hydrophilic treatment, thus providing an attachment environment for microorganisms and impurities and are not convenient for cleaning, they are separated and treated in a targeted manner.

[0093] S4, injecting the initiator into each culture well until the culture well is completely filled, and leaving it to soak for 24 hours, shaking the cell culture plate for 10 seconds every hour at a position deviated from the center of the cell culture plate to make the initiator concentration evenly distributed, and extracting the initiator after the soaking is completed;

[0094] During the static immersion process, the initiator will gradually settle so that the concentration at the bottom is higher than that at the top, which will result in differences in treatment effects. Therefore, the cell culture plate is shaken between treatments to improve the uniformity of treatment.

[0095] S5, spraying the grafting monomer solution onto the cell culture plate and letting it stand for 1 hour. After the reaction is completed, repeatedly rinsing with ethanol solution and deionized water to remove the unreacted initiator and grafting monomer, and drying;

[0096] By reacting the grafting solution with the initiator, a hydrophilic layer can be produced on the inner wall of the culture well of the cell culture plate, while other parts will not be hydrophilized.

[0097] The working principle of the cell culture plate surface treatment device and treatment process will be described in detail below.

[0098] like Figure 1-Figure 14 As shown, a container containing an initiator is placed on the conveying component 1, and the servo module 3 and the first cylinder 5 are controlled to work so that the liquid outlet nozzle 12 extends into the initiator. Initially, the piston plate 27 is in a lower position in the piston cylinder 26, and the valve on the pressure regulating tube 30 is closed. By controlling the second cylinder 25 to work, the piston rod is retracted, so that the piston plate 27 moves upward, thereby drawing negative pressure into the piston cylinder 26, so that the initiator is drawn from the liquid outlet nozzle 12 into the transfer chamber 37.

[0099] The cell culture plate (hereinafter referred to as the culture plate) after acid washing is placed on the conveyor belt. The culture plate is conveyed to the position of the block 34 along with the conveyor component 1 by controlling the conveying component 1 to work. The culture plate is blocked by the block 34 and cannot move further. The lifting plate 6 is pushed downward by controlling the first cylinder 5 to work. The blocking plate 10 faces the cell culture plate. The blocking plate 10 moves downward along with the lifting plate 6 until it is tightly fitted with the cell culture plate. The blocking plate 10 blocks the culture hole into an independent chamber and is not connected with the non-culture hole part.

[0100] Keep the electric valve 14 turned on, and then control the air pump to work. The air pump sucks air into the installation groove 38 through the suction pipe 40, and the suction part is the non-culture hole part. Since this part is attached to the blocking plate 10 and forms a relatively closed chamber, the suction can make the culture plate adsorbed on the blocking plate 10, and the culture plate is taken out;

[0101] In the above process, if the sealing of each culture hole is not good, air leakage will occur. At this time, suction at this part will continuously suck air from the culture hole, so that the initiator in the transfer chamber 37 is sucked into the unsealed culture hole, and further, when the culture hole is full, it will be sucked out of the culture hole; but in this process, when the air pump sucks air to make the initiator enter the culture hole, since the piston plate 27 is controlled by the second cylinder 25 and will not move, the above process will suck air from the external environment through the electric valve 14, and one end of the electric valve 14 has a whistle 15, Therefore, the whistle 15 will sound when the sealing of each culture hole is poor during the adsorption of the culture plate. At the same time, the initiator has infiltrated into the corresponding culture hole. When the operator hears the whistle 15, he will control the air pump to stop vacuuming and release the negative pressure. For a minor sealing problem, the initiator has not yet been pumped to the part outside the culture hole when the whistle 15 is sounded. Then the blocking plate 10 is lifted. At this time, the culture hole with the initiator is the part with the sealing problem, and the operator can deal with the part in a targeted manner.

[0102] In addition, the leakage may also be between the sealing plate 10 and the frame of the culture plate. It is only necessary to stop the vacuuming and let it stand for a few seconds when the culture plate is initially adsorbed. If there is a leakage between the sealing plate 10 and the frame of the culture plate, due to gas leakage, the negative pressure will decrease after a few seconds and the culture plate will not be able to be sucked.

[0103] After the culture plate is sucked up through the above operation, the first cylinder 5 is controlled to be retracted to lift the culture plate, and the piston rod of the second cylinder 25 is controlled to extend to push the piston plate 27 downward, so that the initiator in the transfer chamber 37 is squeezed into each culture hole. During the squeezing process, the air pressure in the culture hole is released through the air outlet 16, and the air pressure is discharged outward after passing through the pressure exhaust pipe 41, the manifold 36 and the manifold 35 in sequence, thereby avoiding the situation where the injection cannot be performed due to the air pressure inside the culture hole;

[0104] With the continuous injection of the initiator, the liquid level in the culture well rises, so that the float 20 floats up and drives the plug 19 and the sliding pin 18 to move upward. When the culture well is filled with the initiator, the plug 19 is completely blocked at this time, so the corresponding culture well can no longer be injected with initiator. At the same time, because the plug 19 blocks the corresponding air outlet 16, the initiator will not overflow. When all the culture wells are filled, the piston plate 27 continues to press down, and there is obviously a large air pressure resistance. When the second cylinder 25 receives the obviously increased resistance, it feeds back a signal to the external controller electrically connected thereto, and the external controller controls the piston rod of the second cylinder 25 to stop extending further, completing the operation of injecting the initiator into the culture well.

[0105] The coordination between the plugging time of the plug 19 and the liquid level inside the culture well is usually achieved by controlling the properties (density and volume) of the float 20, but there may be errors in the actual operation (that is, the above-mentioned precise control is usually only achieved under ideal conditions, and errors need to be considered in the actual operation). Therefore, the float 20 usually pushes the plug 19 to completely block the vent 16 under the action of buoyancy before the culture well is filled, and the remaining small part that has not been immersed needs to be compensated by shaking the culture plate, specifically:

[0106] By controlling the motor 7 to work, the rocker 32 is driven to rotate, and further, the cover 8 and the structure connected thereto are caused to rotate around the output shaft of the motor 7; and the cover 8 is limited by the adapter block 28 and the adapter 29 at the same time, so the direction will not change, but the rotation is completed without changing the direction, thereby achieving shaking, so that the initiator can move along the inner wall of the culture hole; the immersion effect of the part near the top of the culture hole is worse than that of the bottom (based on the situation that the initiator is not completely filled), so the hydrophilicity at the top of the hole is weaker than that at the bottom, which can inhibit the adherent cells from extending out of the culture hole;

[0107] In the above process, the rocker 32 will rub against the annular plate 31 during its rotation, and the contact surfaces of the two are rough surfaces, so high-frequency jitter will be generated during the relative friction, thereby making the initiator concentration distribution more uniform and easier to diffuse to various parts.

[0108] After the soaking is completed, the second cylinder 25 is controlled to be retracted, driving the piston plate 27 to move upward, thereby applying negative pressure to the piston cylinder 26, so that the initiator in the culture well is sucked into the transfer chamber 37, and can be used multiple times;

[0109] Then control the motor 7 to stop working and reset to the initial state, then control the servo module 3 to work and transport the culture plate to the other side of the block 34, and control the first cylinder 5 to extend so that the culture plate is placed on the conveyor belt, and then control the air pump to stop supplying air and release the negative pressure, so that the culture plate can be released and transported to the spraying part 2 along with the conveying part 1. The spraying part 2 sprays the grafting solution on the soaked cell culture plate and leaves it to stand for 1 hour. After the reaction is completed, it is repeatedly rinsed with ethanol solution and deionized water to remove the unreacted initiator and grafting monomer, and dried to obtain a hydrophilic cell culture plate. In the above operation, only the culture hole part of the cell culture plate undergoes the entire process operation, and the parts outside the culture hole are not subjected to chemical soaking (initiator) operation, so hydrophilicity will not be obtained.

[0110] The above generally describes the present invention in detail, but it is obvious to a person skilled in the art that some modifications or improvements can be made to the present invention. Therefore, modifications or improvements that do not depart from the spirit of the present invention are within the scope of protection of the present invention.

Claims

1. A cell culture plate surface treatment device, comprising: A conveying component (1), the conveying component (1) being used for conveying a cell culture plate, the conveying component (1) comprising a base and a conveying belt; a stopper (34) is fixedly connected to the inner side of the base; An immersion component, wherein the immersion component is used to chemically immerse the cell culture plate; A spraying component (2), the spraying component (2) being used to spray the grafting monomer solution onto the cell culture plate after chemical immersion; The immersion component comprises a transfer plate (9), a mounting groove (38) is provided at the bottom of the transfer plate (9), a sealing plate (10) is connected to the inside of the mounting groove (38) by screws, the sealing plate (10) is fitted with the inner wall of the mounting groove (38), the top and bottom of the sealing plate (10) are fixedly connected with sealing strips, the sealing strip at the top of the sealing plate (10) is tightly fitted with the mounting groove (38), the sealing plate (10) is squeezed and matched with the cell culture plate to separate each culture well into a closed chamber, and the top of the transfer plate (9) is fixedly connected with a cover shell (8); A transfer cavity (37) is formed at the top of the blocking plate (10); a plurality of liquid outlet nozzles (12) are fixedly connected to the bottom of the blocking plate (10); a through hole for connecting the liquid outlet nozzles (12) with the transfer cavity (37) is formed on the blocking plate (10); an input pipe (39) connected to the mounting groove (38) is fixedly connected to the top of the transfer plate (9); a plurality of sealing rings (13) are fixedly connected to the bottom of the blocking plate (10); the sealing rings (13) correspond one to one to the liquid outlet nozzles (12); It also includes: a transport component, which is arranged outside the conveying component (1) and is used to take the cell culture plate and shake it; An injection component is arranged above the cover (8) and is used to provide an initiator to the immersion component through the input pipe (39).

2. A cell culture plate surface treatment device according to claim 1, characterized in that: The transfer component comprises a servo module (3) symmetrically mounted on both sides of a base of the conveying component (1); a movable end of the servo module (3) is fixedly connected to a U-shaped frame (4); a first cylinder (5) is fixedly connected to the top of the inner side of the U-shaped frame (4); a piston rod of the first cylinder (5) is fixedly connected to a lifting plate (6); a motor (7) is fixedly connected to the top of the lifting plate (6); an output end of the motor (7) extends to the bottom of the lifting plate (6) and is fixedly connected to a rocker (32); and the cover (8) is rotatably connected to the bottom of the rocker (32); The two sides of the cover shell (8) are symmetrically fixedly connected to the extension plates (11); the bottom of the lifting plate (6) is rotatably connected to the adapter block (28), the bottom of the adapter block (28) is rotatably connected to the adapter (29), and the bottom of the adapter (29) is rotatably connected to the extension plate (11); the top of the transfer plate (9) is fixedly connected to the suction pipe (40), and the transfer plate (9) is provided with a through hole connecting the suction pipe (40) and the non-culture hole part of the cell culture plate; one side of the base is fixedly connected to the air pump, and the air pump is connected to the suction pipe (40).

3. A cell culture plate surface treatment device according to claim 2, characterized in that: A plurality of collecting bars (36) are fixedly connected to the top of the transfer plate (9), and a collecting pipe (35) is commonly connected to the tops of the collecting bars (36). An electric valve (14) is fixedly connected to one side of the transfer plate (9), and one end of the collecting pipe (35) is connected to the electric valve (14); the other end of the electric valve (14) is connected to a whistle (15); a plurality of pressure-discharging pipes (41) are fixedly connected to the bottom of the inner wall of the transfer chamber (37); a plurality of through holes corresponding to the pressure-discharging pipes (41) are opened at the top of the inner wall of the mounting groove (38), and the through holes are connected to the collecting bars (36); and an air outlet hole (16) connected to the pressure-discharging pipe (41) is opened at the bottom of the blocking plate (10).

4. A cell culture plate surface treatment device according to claim 3, characterized in that: A transverse plate (17) is fixedly connected between the inner walls of the air outlet holes (16); a slide rod (21) is slidably connected to the bottom of the transverse plate (17); a plug (19) is fixedly connected to the bottom of the slide rod (21); a float (20) is fixedly connected to the bottom of the plug (19); a slide pin (18) is fixedly connected to the outer side of the slide rod (21); the top of the slide rod (21) extends to the top of the transverse plate (17) and is fixedly connected to a limit plate (22); and a plurality of strip grooves are provided on the outer side of the slide pin (18).

5. A cell culture plate surface treatment device according to claim 4, characterized in that: The injection component comprises a mounting frame (24), wherein the mounting frame (24) is fixedly connected to the top of the lifting plate (6), a second cylinder (25) is fixedly connected to the top of the mounting frame (24), a piston cylinder (26) is fixedly connected to the inner side of the mounting frame (24), a piston plate (27) is slidably connected between the inner walls of the piston cylinder (26), a piston rod of the second cylinder (25) extends into the interior of the piston cylinder (26) and is fixedly connected to the piston plate (27); a pressure regulating pipe (30) is provided on the piston cylinder (26), a valve is provided on the pressure regulating pipe (30), and the bottom of the piston cylinder (26) is communicated with an input pipe (39).

6. A cell culture plate surface treatment device according to claim 5, characterized in that: A force-bearing plate (33) is fixedly connected to the inner side of the base, and the top of the conveyor belt is higher than the top of the force-bearing plate (33).

7. A cell culture plate surface treatment device according to claim 6, characterized in that: The interior of the liquid outlet nozzle (12) is provided with a honeycomb hole structure (23).

8. A cell culture plate surface treatment device according to claim 7, characterized in that: An annular plate (31) is fixedly connected to the bottom of the lifting plate (6), and the bottom of the annular plate (31) and the top of the rocker (32) are both arranged as rough surfaces and in contact with each other.

9. A cell culture plate surface treatment process, characterized in that: The cell culture plate surface treatment device applicable to any one of claims 1 to 8 above comprises the following steps: S1. Soak the cell culture plate in anhydrous ethanol and then rinse it repeatedly with deionized water; S2. Completely immerse the cleaned cell culture plate in a sulfuric acid-nitric acid mixed acid solution at 25-40° C. for 4-6 hours, take it out, wash it repeatedly with deionized water, and dry it; S3, blocking the top of the cell culture plate so that each culture well is separated into a relatively closed chamber; S4, injecting the initiator into each culture well until the culture well is completely filled, and leaving it to soak for 24 hours, shaking the cell culture plate for 10 seconds every hour at a position deviated from the center of the cell culture plate to make the initiator concentration evenly distributed, and extracting the initiator after the soaking is completed; S5. Spray the grafting monomer solution onto the cell culture plate and let it stand for 1 to 1.5 hours. After the reaction is completed, rinse it repeatedly with ethanol solution and deionized water to remove the unreacted initiator and grafting monomer, and dry it.

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