CN114181832A - Auxiliary device convenient for operating NK cell in-vitro separation culture and use method - Google Patents
Auxiliary device convenient for operating NK cell in-vitro separation culture and use method Download PDFInfo
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Abstract
The invention discloses an auxiliary device convenient for operating NK cell in-vitro separation culture and a using method, and particularly relates to the technical field of NK cell in-vitro separation culture, which comprises an incubator, wherein a plurality of positioning groove plates are sequentially arranged from top to bottom in the incubator, the top ends of the positioning groove plates are connected with a linkage support plate in a sliding manner, and one side of the linkage support plate is provided with an inspection mechanism; the inspection mechanism comprises a movable thread block arranged on one side of the linkage support plate, and a driving screw is arranged in the movable thread block. According to the invention, the inspection mechanism is adopted, and the NK cells in each culture tank in the incubator can be checked up and down without opening the incubator, so that the effect of three-axis linkage checking is realized, the NK cell culture condition is accurately mastered, the culture accuracy is improved, the culture condition in the incubator cannot be influenced by external factors, the culture accuracy is improved, and the culture effect is better.
Description
Technical Field
The invention relates to the technical field of NK cell in-vitro separation culture, in particular to an auxiliary device for NK cell in-vitro separation culture convenient to operate and a using method.
Background
NK cells are natural killer cells are important immune cells of the body, not only associated with anti-tumor, anti-viral infection and immune regulation, but also involved in the development of hypersensitivity and autoimmune diseases in some cases, since NK cells have no MHC restriction in their killing activity, do not depend on antibodies, and are called natural killer activity. The NK cell is rich in cytoplasm, contains large azurophilic granules, and the content of the granules is positively correlated with the killing activity of the NK cell. The killing effect of the NK cells after acting on the target cells appears early, the killing effect can be seen in 1 hour in vitro and 4 hours in vivo, and an auxiliary device is required to be used for carrying out auxiliary culture operation in the separation culture process of the NK cells.
In actual use, culture solution needs to be placed in the NK cell culture dish for long-time culture, the culture dish is placed in the incubator to complete culture operation, the NK cell culture condition in each culture dish needs to be opened periodically, and data is recorded in time, so that the following defects exist;
need open inside the looking over of whole incubator and cultivate the cell situation, can cause incubator internal culture's humidity, temperature, change like this to and contact outside air humidity and so on factor can lead to the inside cell culture of a plurality of culture dishes to influence great, and it is relatively poor to cultivate the effect, and need take out the culture dish during the observation and look over, and the viewing time is longer, operates very inconvenient and look over the accuracy relatively poor.
Disclosure of Invention
In order to overcome the defects in the prior art, the embodiment of the invention provides the auxiliary device for conveniently operating the in-vitro separation and culture of the NK cells and the using method thereof.
In order to achieve the purpose, the invention provides the following technical scheme: an auxiliary device convenient for operating NK cell in-vitro separation culture comprises an incubator, wherein a plurality of positioning groove plates are sequentially arranged from top to bottom in the incubator, a linkage support plate is connected to the top end of each positioning groove plate in a sliding mode, and an inspection mechanism is installed on one side of each linkage support plate;
the inspection mechanism is including setting up the removal screw thread piece in linkage extension board one side, remove the inside drive screw that is provided with of screw thread piece, the dwang is installed in linkage extension board bottom embedding, the dwang is outside and be located the linkage extension board about the equal symmetrical cover in both sides be equipped with cup joint the frame board, cup joint the frame inboard wall and be connected with the linkage screw, the outside cover of linkage screw is equipped with removes the spiral shell piece, remove spiral shell piece bottom and install the camera, linkage screw one end is installed and is used for driven first driving motor.
In a preferred embodiment, one side of the sleeving frame plate is connected with a linkage block, one side of the linkage block is connected with a movable belt, a first belt pulley is sleeved in the movable belt, a second driving motor is installed at one end of the first belt pulley, a second belt pulley is arranged on one side of the first belt pulley, a rotating shaft lever is arranged in the second belt pulley, a display is arranged on one side of the incubator, a rotating motor is arranged at the top end of the rotating rod, a moving motor is arranged at the top end of the driving screw rod, the interior of the movable screw block is in threaded connection with the exterior of the driving screw, the interior of the movable screw block is in threaded connection with the exterior of the linkage screw, the outer part of the rotating rod and the position between the two sleeve frame plates are connected with the inner part of the linkage support plate in a sliding way, the dwang outside with cup joint frame board fixed connection, a plurality of culture tanks have been seted up on transparent culture plate top.
In a preferred embodiment, the other side of the sleeve frame plate is connected with three connecting support rods, one end of each connecting support rod is fixedly connected with a culture mechanism, the culture mechanism comprises an injection pipe fixedly connected with one end of the connecting support rod, three atomizing injection heads are installed below the outer wall of the injection pipe, a connecting pipe is communicated above the outer wall, a first electric control valve is installed at the top end of the connecting pipe, a flow meter is installed at the top end of the first electric control valve, one end of the flow meter is connected with a first shunt pipe, a connecting hose for stretching is communicated above the outer wall of the first shunt pipe, the top end of the connecting hose is connected with an injection pipe, an injection pump machine for conveying culture solution is installed at the bottom end of the injection pipe, the input end of the injection pump machine is communicated with a second shunt pipe, a second electric control valve is installed at one end of the second shunt pipe, and a first container is installed at one end of the second electric control valve, and the other end of the second shunt pipe is provided with a third electric control valve, the bottom end of the third electric control valve is provided with a second container, the three atomizing spray heads are communicated with a connecting pipe, and two ends of the connecting hose are respectively communicated with the first shunt pipe and the injection pipe in pairs.
In a preferred embodiment, two hot air diffusion pipes are installed to the culture tank top, the hot air diffusion pipe top is connected with the third shunt pipe, third shunt pipe top intercommunication has the input tube, input tube one end is provided with the reposition of redundant personnel injection pipe, and reposition of redundant personnel injection pipe outer wall one side intercommunication has the booster pipe, circulating fan is installed to the booster pipe bottom, circulating fan input intercommunication has the circulation case, circulation case internally mounted has condenser tube, circulation outlet pipe and the other end are installed to condenser tube one end and are installed the circulation inlet tube, circulation inlet tube top is provided with the suction tube, suction tube one end is connected with the low straw of diffusion, the electrical heating board is installed to the circulation case bottom, suction tube is linked together with the low straw of diffusion.
A use method of an auxiliary device for conveniently operating NK cell in-vitro separation and culture comprises the following specific use steps:
step one, during temperature circulation, putting NK cell culture raw materials into a culture tank on each transparent culture plate, then aligning the transparent culture plates to two positioning groove plates for sliding insertion, closing a sealing door of the culture box, heating air in the circulation box by starting an electric heating plate when the temperature is low, monitoring the interior of the circulation box by using a temperature measuring instrument, starting a circulating fan when the temperature reaches the NK cell culture temperature so as to enable a diffusion low suction pipe to suck the air in the culture box into a cooling water pipe, enabling the cooling water pipe to enter the circulation box, entering a pressure boost pipe along the circulation box, entering a shunt injection pipe from the pressure boost pipe, entering a plurality of input pipes through the shunt injection pipe, injecting into a third shunt pipe through the input pipe, entering a hot air diffusion pipe through the third shunt pipe for uniform downward blowing, multiple layers are formed, the multiple layers are blown from top to bottom, then the multiple layers enter three through holes in the middle of a transparent culture plate to move downwards, hot air uniformly covers the upper surface of the transparent culture plate and flows to the lower surface of the transparent culture plate, then the hot air enters a diffusion low suction pipe to circulate into a suction pipe, thermal circulation is formed, when the temperature is hot, an electric heating plate can be closed, a circulation outlet pipe is connected with a water return pipeline of a cooling water tower, a circulation water inlet pipe is connected with a water inlet pipeline of the cooling water tower, cooling water can be injected into a bucket cooling water pipe, the air in a circulation box is cooled, the effect of cooling is achieved, cooling air is circulated into the culture box by a circulation fan, and the temperature can be adjusted through cold and hot circulation;
step two, when checking the culture condition, starting a rotating motor to drive a rotating rod to rotate ninety degrees clockwise, driving two rotating rods to rotate, sleeving a frame plate to drive a linkage screw rod to enable a movable screw block to rotate, driving a camera to move to the rightmost position of the upper surface of a transparent culture plate and moving another camera to the lower surface of the transparent culture plate, starting a first driving motor to drive the linkage screw rod to rotate forwards, driving the movable screw block to move backwards under the action of threads by the linkage screw rod, driving the camera to move backwards along the upper surface of the transparent culture plate by the movable screw block, thus playing a role of checking and monitoring the upper surfaces of three culture tanks, inputting the checked images into a display to display, checking the culture condition of NK cells at the bottom end of the inner wall of the culture tank by the camera below the transparent culture plate, and penetrating image data into the display to perform double-image display, and the second driving motor is started to drive the first belt pulley to rotate forwards, the first belt pulley drives the moving belt to transmit, and simultaneously the second belt pulley outside the shaft lever is rotated to transmit, the moving belt can drive the linkage block to move rightwards, the linkage block drives the moving thread block to move rightwards, the moving thread block drives the rotating rod to enable the two sleeving frame plates to move rightwards, so that the sleeving frame plates drive the cameras inside to move rightwards to three culture tanks at the other side position, then the first driving motor is started to drive the linkage screw rod to rotate backwards, the linkage screw rod drives the moving thread block to enable the cameras to move forwards, the culture conditions in the other three culture tanks can be continuously checked, after the cameras can move rightwards to a certain distance each time, the cameras can move forwards and backwards to check the culture conditions in the culture tanks, after the culture tanks on the transparent culture plate below are checked completely, starting a second driving motor to drive a first belt pulley to rotate reversely, and transmitting until the sleeved frame plate is reset to the leftmost position of the inner wall of the incubator, then starting a rotating motor to drive a rotating rod to rotate anticlockwise and reset to the initial position, so that the sleeved frame plate can be reset to the initial position, then starting a moving motor to drive a driving screw to rotate forwards, and driving the driving screw to drive a moving thread block to move upwards under the action of threads to check the culture tank in the transparent culture plate on the upper layer, so that the multilayer transparent culture plates can be checked repeatedly;
step three, when the culture solution is added, in the observation process, the sleeve frame plate can drive the connecting support rod to move, the connecting support rod drives the injection pipe to move, so that the injection pipe moves along the upper surface of the culture tank, and the three culture tanks are butted by the three groups of atomizing injection heads, so that in the observation process, if the NK cell culture solution is less, serum-free complete culture medium liquid in the second containing box or autologous plasma in the first containing box can be selected, when the culture solution in the first containing box is selected, the third electric control valve can be closed, the second electric control valve is opened, when the serum-free complete culture medium liquid in the second containing box is selected, the second electric control valve can be closed, the second electric control valve is opened, so that two different culture solutions can be selected, when the culture solution in the first containing box is selected, the injection pump can be started, the autologous plasma culture fluid in the first containing box enters the injection tube, is injected into the connecting hose through the injection tube and enters the first shunt tube along the connecting hose, the liquid is divided by the first shunt pipe and enters the opened first electric control valve through the flowmeter, is guided into the connecting pipe by the first electric control valve and enters the spray pipe, and is finally sprayed into the culture tank through the atomizing spray head, thus, the culture solution can be uniformly atomized and sprayed at one time, and when the culture solution in one of the culture tanks is less in the observation process of the camera, the injection quantity can be set, the other two first electric control valves are closed, the metering is carried out through the flow meter, when the amount of the liquid to be sprayed reaches a predetermined amount, the connection pipe is closed, so that the operation of controlling the liquid culture medium can be performed in any one of the culture tanks, thereby achieving a high-precision culture operation.
The invention has the technical effects and advantages that:
1. the invention adopts an inspection mechanism to start a rotating motor to drive a rotating rod to rotate ninety degrees clockwise, a frame plate is sleeved to drive a linkage screw rod to drive a movable screw block to rotate, a camera head moves to the rightmost position of the upper surface of a transparent culture plate, another camera head moves to the lower surface of the transparent culture plate, a first drive motor drives the linkage screw rod to drive the movable screw block to move backwards under the action of screw threads, the camera head on the upper surface can play a role in checking and monitoring the upper surfaces of three culture tanks, the camera head below the transparent culture plate can check the culture condition of NK cells at the bottom end of the inner wall of the culture tank, image data penetrates into a display to be displayed in a double-image mode, a second drive motor drives a first belt pulley to rotate forwards, a moving belt can drive the linkage block to move rightwards to check, and after one layer is checked, the drive screw rod can also be driven to rotate forwards through the moving motor, the effect of upward moving can be realized, so that the NK cells in each culture tank in the incubator can be checked up and down without opening the incubator, the checking is more convenient, the effect of three-axis linkage checking is realized, the NK cell culture condition is accurately mastered, the culture accuracy is improved, the external factors cannot influence the culture condition in the incubator, the culture accuracy is improved, and the culture effect is better;
2. the invention adopts a culture mechanism which can select serum-free complete culture medium liquid in a second containing box or autologous plasma in a first containing box, starts an injection pump machine to enable autologous plasma culture liquid in the first containing box to enter an injection pipe, and then the autologous plasma culture liquid enters an opened first electric control valve through a first shunt pipe shunt belt flow meter, is guided into a connecting pipe to enter an injection pipe, and is downwards sprayed into a culture tank through an atomizing spray head, so that the culture liquid can be uniformly atomized and sprayed at one time, when the culture liquid in one culture tank is less in the observation process of a camera, the spray volume can be set, the other two first electric control valves are closed, and the culture liquid is downwards sprayed through the flow meter in a metering way, so that the switching spray of various culture liquids can be formed, and the addition operation of the culture liquid volume can be carried out according to the actual culture condition, the addition operation can be carried out on different culture solutions, the culture solution can be accurately mastered and used for carrying out the culture operation, so that the NK cells can be subjected to the accurate culture solution culture operation to the greatest extent, the culture solution is not wasted, the accurate culture operation in each culture tank can be realized, and the culture effect is improved;
3. when the temperature reaches the NK cell culture temperature, the circulating fan is adopted to enable the diffusion low suction pipe to suck the air in the incubator into the cooling water pipe, the air is input into the shunt injection pipes from the pressure increasing pipe and enters the multiple input pipes, the air enters the hot air diffusion pipe through the third shunt pipe to be uniformly blown downwards, the hot air uniformly covers and flows on the upper surface of the transparent culture plate and then flows to the lower surface of the transparent culture plate, and then enters the diffusion low suction pipe to be circulated into the suction pipe, so that the uniform heating or temperature cooling of the air in the incubator can be realized, the upper surface of each transparent culture plate can be uniformly covered and flows in the circulating process, the requirement on uniform temperature culture is met, the accurate temperature covering culture operation is achieved, and the culture effect is better;
in conclusion, through the mutual influence of the above functions, the function of three-axis linkage checking is realized, the culture condition of NK cells is accurately mastered, the culture accuracy is improved, and will not cause the external factor to influence the internal culture situation of the incubator, improve the culture accuracy, the culture effect is better, the NK cells can be performed with the precise culture solution culture operation to the maximum extent, not only the culture solution can not be wasted, but also can achieve the accurate culture operation in each culture tank, improve the culture effect, promote the temperature covering flow on the upper surface of each transparent culture plate in the circulating process, achieve the better uniform temperature culture requirement, achieve the accurate temperature covering culture operation, and in conclusion, can realize the high-accuracy culture operation in the process of culturing NK cells, the inside of each culture tank is precisely cultured, so that the culture effect is effectively improved, and the culture quality is ensured.
Drawings
Fig. 1 is a schematic view of the overall structure of the present invention.
FIG. 2 is a schematic view of the sectional structure of the driving screw according to the present invention.
FIG. 3 is a schematic view of the structure of the joint between the movable screw block and the linkage support plate according to the present invention.
Fig. 4 is an enlarged schematic view of a portion a in fig. 3 according to the present invention.
Fig. 5 is a schematic structural view of a second pulley of the present invention.
Fig. 6 is an enlarged schematic view of the structure at B in fig. 5 according to the present invention.
FIG. 7 is a schematic view showing the internal structure of the incubator and the circulation box according to the present invention.
FIG. 8 is a schematic view of the structure of the connection between the atomizing spray head and the spray tube of the present invention.
Fig. 9 is a rear perspective view of the present invention.
The reference signs are: 1. an incubator; 2. positioning the groove plate; 3. a transparent culture plate; 4. a linkage support plate; 5. moving the thread block; 6. a drive screw; 7. rotating the rod; 8. sleeving a frame plate; 9. a linkage screw; 10. moving the screw block; 11. a first drive motor; 12. a camera; 13. a linkage block; 14. moving the belt; 15. a first pulley; 16. a second drive motor; 17. a second pulley; 18. rotating the shaft lever; 19. a rotating electric machine; 20. a moving motor; 21. a culture tank; 22. connecting a support rod; 23. an injection pipe; 24. an atomizing spray head; 25. a connecting pipe; 26. a first electrically controlled valve; 27. a flow meter; 28. a first shunt pipe; 29. a connecting hose; 30. an injection pipe; 31. an injection pump machine; 32. a second shunt pipe; 33. a second electrically controlled valve; 34. a first container box; 35. a third electrically controlled valve; 36. a second container box; 37. a hot air diffusion pipe; 38. a third shunt pipe; 39. an input tube; 40. a shunt injection pipe; 41. a pressure increasing pipe; 42. a circulating fan; 43. a circulation box; 44. a cooling water pipe; 45. a circulation outlet pipe; 46. circulating a water inlet pipe; 47. a suction tube; 48. a diffusion low suction pipe; 49. an electrical heating plate; 50. a display.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The auxiliary device for conveniently operating the in-vitro NK cell separation and culture comprises an incubator 1, wherein a plurality of positioning groove plates 2 are sequentially arranged from top to bottom in the incubator 1, the top ends of the positioning groove plates 2 are slidably connected with a linkage support plate 4, and an inspection mechanism is arranged on one side of the linkage support plate 4;
inspection mechanism is including setting up the removal screw thread piece 5 in linkage extension board 4 one side, remove the inside drive screw 6 that is provided with of screw thread piece 5, dwang 7 is installed in the embedding of linkage extension board 4 bottom, dwang 7 outside just is located linkage extension board 4 about the equal symmetrical cover in both sides be equipped with cup joint deckle board 8, cup joint deckle board 8 inner wall connection has linkage screw 9, the outside cover of linkage screw 9 is equipped with removes spiral shell piece 10, remove spiral shell piece 10 bottom and install camera 12, linkage screw 9 one end is installed and is used for driven first driving motor 11.
In some embodiments, as shown in fig. 1-7, one side of the sleeved frame plate 8 is connected with a linkage block 13, one side of the linkage block 13 is connected with a movable belt 14, the movable belt 14 is internally sleeved with a first belt pulley 15, one end of the first belt pulley 15 is provided with a second driving motor 16, one side of the first belt pulley 15 is provided with a second belt pulley 17, the second belt pulley 17 is internally provided with a rotating shaft 18, one side of the incubator 1 is provided with a display 50, the top end of the rotating rod 7 is provided with a rotating motor 19, the top end of the driving screw 6 is provided with a movable motor 20, the inside of the movable screw block 5 is in threaded connection with the outside of the driving screw 6, the inside of the movable screw block 10 is in threaded connection with the outside of the linkage screw 9, the outside of the rotating rod 7 and located between the two sleeved frame plates 8 is in sliding connection with the inside of the linkage support plate 4, the outside of the rotating rod 7 is fixedly connected with the sleeved frame plates 8, the top end of the transparent incubator 3 is provided with a plurality of culture grooves 21, so that second driving motor 16 drives first belt pulley 15 corotation, and first belt pulley 15 drives and removes belt 14 and carries out the transmission, rotates the outside second belt pulley 17 of axostylus axostyle 18 and carries out the transmission, removes belt 14 and can drive linkage piece 13 and move to the right, cup joints the operation that the deckle board 8 drove its inside camera 12 and move to the right, realizes lateral shifting.
In some embodiments, as shown in fig. 4-9, the other side of the sleeved frame plate 8 is connected with three connecting support rods 22, one end of each connecting support rod 22 is fixedly connected with a culture mechanism, the culture mechanism comprises an injection pipe 23 fixedly connected with one end of the connecting support rod 22, three atomizing and spraying heads 24 are installed below the outer wall of the injection pipe 23, a connecting pipe 25 is communicated with the upper part of the outer wall, a first electric control valve 26 is installed at the top end of the connecting pipe 25, a flow meter 27 is installed at the top end of the first electric control valve 26, a first shunt pipe 28 is connected with one end of the flow meter 27, a connecting hose 29 for stretching is communicated with the upper part of the outer wall of the first shunt pipe 28, an injection pipe 30 is connected with the top end of the connecting hose 29, an injection pump 31 for conveying culture solution is installed at the bottom end of the injection pipe 30, the input end of the injection pump 31 is communicated with a second shunt pipe 32, a second electric control valve 33 is installed at one end of the second shunt pipe 32, a first holding box 34 is arranged at one end of the second electric control valve 33, a third electric control valve 35 is arranged at the other end of the second shunt pipe 32, a second holding box 36 is arranged at the bottom end of the third electric control valve 35, the three atomizing and spraying heads 24 are communicated with the connecting pipe 25, two ends of the connecting hose 29 are respectively communicated with the first shunt pipe 28 and the injection pipe 30 in pairs, so that serum-free complete culture medium liquid in the second holding box 36 or autologous plasma in the first holding box 34 can be ensured, the autologous plasma culture liquid in the first holding box 34 can enter the injection pipe 30 and enter the first shunt pipe 28 along the connecting hose 29, the autologous plasma culture liquid is guided into the connecting pipe 25 through the first shunt pipe 28 and is delivered into the spraying pipe 23, and finally is downwards sprayed into the culture tank 21 through the atomizing and spraying head 24, when the culture liquid in one of the culture tanks 21 is found to be less in the observation process of the camera 12, the other two first electrically controlled valves 26 are closed and metering is performed by the flow meter 27, which makes it possible to form and refine the culture operation.
In some embodiments, as shown in fig. 2-7, two hot air diffusion pipes 37 are installed above the cultivation tank 21, the top end of each hot air diffusion pipe 37 is connected with a third shunt pipe 38, the top end of the third shunt pipe 38 is communicated with an input pipe 39, one end of the input pipe 39 is provided with a shunt injection pipe 40, one side of the outer wall of the shunt injection pipe 40 is communicated with a pressure increasing pipe 41, the bottom end of the pressure increasing pipe 41 is provided with a circulating fan 42, the input end of the circulating fan 42 is communicated with a circulating box 43, the inside of the circulating box 43 is provided with a cooling water pipe 44, one end of the cooling water pipe 44 is provided with a circulating outlet pipe 45, the other end of the cooling water pipe is provided with a circulating inlet pipe 46, the upper part of the circulating inlet pipe 46 is provided with a suction pipe 47, one end of the suction pipe 47 is connected with a diffusion low suction pipe 48, the bottom end of the circulating box 43 is provided with an electric heating plate 49, the suction pipe 47 is communicated with the diffusion low suction pipe 48 so as to start the electric heating plate 49 to heat the air inside the circulating box 43, thereby circulating fan 42 makes diffusion low suction pipe 48 with incubator 1 inside air suction to condenser tube 44 in, circulation case 43 enters into pressure increasing pipe 41, input into reposition of redundant personnel injection pipe 40 by pressure increasing pipe 41 in, third reposition of redundant personnel 38 enters into hot-blast diffusion tube 37 and evenly blows down, hot gas evenly covers and flows to transparent cultivation board 3 lower surface at transparent cultivation board 3 upper surface, form thermal cycle, circulation outlet pipe 45 connects cooling tower's return water pipeline, and circulation inlet pipe 46 connects cooling tower's inlet channel can be with cooling water injection fill condenser tube 44 in, form the cold cycle operation, realize the balanced cover formula of temperature fluidization.
A use method of an auxiliary device for conveniently operating NK cell in-vitro separation and culture comprises the following specific use steps:
step one, during temperature circulation, putting NK cell culture raw materials into a culture groove 21 on each transparent culture plate 3, then inserting the transparent culture plate 3 into two positioning groove plates 2 in a sliding way, closing a sealing door of the culture box 1, heating air inside a circulation box 43 by starting an electric heating plate 49 when the temperature is low, monitoring the inside of the circulation box 43 by using a temperature measuring instrument, starting a circulation fan 42 when the temperature reaches the NK cell culture temperature, enabling a diffusion low suction pipe 48 to suck the air inside the culture box 1 into a cooling water pipe 44, enabling the cooling water pipe 44 to enter the circulation box 43, entering a pressure boosting pipe 41 along the circulation box 43, inputting the air into a shunt injection pipe 40 by the pressure boosting pipe 41 and inputting the air into a plurality of input pipes 39 by the shunt injection pipe 40, and injecting the air into a third shunt pipe 38 by the input pipe 39, and enters the hot air diffusion pipe 37 through the third branch pipe 38 to be uniformly blown down, and is formed into a plurality of layers from top to bottom, then the hot air enters into three through holes in the middle of the transparent culture plate 3 to move downwards, and the hot air uniformly covers and flows on the upper surface of the transparent culture plate 3 and then flows to the lower surface of the transparent culture plate 3, then enters the diffusion low suction pipe 48 to circulate to the suction pipe 47, and forms a thermal cycle, and when the temperature is hot, the cooling water can be cooled by turning off the electric heating plate 49, and connecting the circulation outlet pipe 45 to the return pipe of the cooling water tower, and the circulating water inlet pipe 46 is connected to the water inlet pipe of the cooling water tower to inject the cooling water into the bucket cooling water pipe 44, thus cooling the air inside the circulating box 43, thereby playing a role of cooling, and the cooling air is circulated into the incubator 1 by the circulating fan 42, so that the temperature can be adjusted by cold and hot circulation;
step two, when checking the culture condition, starting the rotating motor 19 to drive the rotating rod 7 to rotate ninety degrees clockwise, driving two rotating rods 7 to rotate, sleeving the frame plate 8 to drive the linkage screw 9 to rotate the movable screw block 10, driving the camera 12 to move to the position of the other camera 12 at the rightmost position of the upper surface of the transparent culture plate 3 to move to the position of the lower surface of the transparent culture plate 3 by the movable screw block 10, starting the first driving motor 11 to drive the linkage screw 9 to rotate forwardly, driving the movable screw block 10 to move backwardly under the action of the screw thread by the linkage screw 9, driving the camera 12 to move backwardly along the upper surface of the transparent culture plate 3 by the movable screw block 10, thus playing a role of checking and monitoring the upper surfaces of the three culture tanks 21, inputting the checked image to the display 50 to display, and checking the culture condition of NK cells at the bottom end of the inner wall of the culture tanks 21 by the camera 12 below the transparent culture plate 3, image data is penetrated into a display 50 to carry out double-image display, a second driving motor 16 is started to drive a first belt pulley 15 to rotate forwards, the first belt pulley 15 drives a moving belt 14 to carry out transmission, meanwhile, a second belt pulley 17 outside a shaft lever 18 is rotated to carry out transmission, the moving belt 14 can drive a linkage block 13 to move rightwards, the linkage block 13 drives a moving threaded block 5 to move rightwards, the moving threaded block 5 drives a rotating lever 7 to move rightwards so that two sleeve frame plates 8 move rightwards, thus, the sleeve frame plates 8 drive a camera 12 inside the sleeve frame plates to move rightwards to three culture tanks 21 at the other side position, then a first driving motor 11 is started to drive a linkage screw 9 to rotate backwards, the linkage screw 9 drives a moving threaded block 10 to move the camera 12 forwards, the other three culture tanks 21 can be continuously checked for culture conditions, and after a distance is reached each time, the camera 12 can move back and forth to check the culture condition in the culture tank 21, after the culture tank 21 on the lower transparent culture plate 3 is checked, the second driving motor 16 is started to drive the first belt pulley 15 to rotate reversely and transmit until the sleeve frame plate 8 is reset to the leftmost position on the inner wall of the culture box 1, then the rotating motor 19 is started to drive the rotating rod 7 to rotate anticlockwise and reset to the initial position, so that the sleeve frame plate 8 is reset to the initial position, then the moving motor 20 is started to drive the driving screw 6 to rotate forwards, the driving screw 6 drives the moving thread block 5 to move upwards under the action of the threads to check the culture tank 21 in the upper transparent culture plate 3, and thus, the multilayer transparent culture plates 3 can be checked repeatedly;
step three, during the culture solution addition, during the viewing process, the connecting support rod 22 is driven by the sleeving frame plate 8 to move, the connecting support rod 22 drives the injection pipe 23 to move, so that the injection pipe 23 moves along the upper surface of the culture tank 21, and the three culture tanks 21 are butted by the three groups of atomizing injection heads 24, so that during the viewing process, if the NK cell culture solution is less, serum-free complete culture medium liquid inside the second container box 36 or autologous plasma inside the first container box 34 can be selected, when the culture solution inside the first container box 34 is selected, the third electric control valve 35 can be closed, the second electric control valve 33 is opened, when the serum-free complete culture medium liquid inside the second container box 36 is selected, the second electric control valve 33 can be closed, the second electric control valve 33 is opened, so that two different culture solutions can be selected, when the culture fluid in the first container 34 is selected, the injection pump 31 is started, the autologous plasma culture fluid in the first container 34 enters the injection pipe 30, is injected into the connecting hose 29 through the injection pipe 30, enters the first shunt pipe 28 along the connecting hose 29, is shunted by the first shunt pipe 28 and carries the flow meter 27, enters the opened first electric control valve 26 through the flow meter 27, is guided into the connecting pipe 25 through the first electric control valve 26 and enters the injection pipe 23, and is finally injected into the culture tank 21 through the atomizing injection head 24, so that the culture fluid can be uniformly atomized and injected at one time, when the culture fluid in one of the culture tanks 21 is less in the observation process of the camera 12, the injection amount can be set, the other two first electric control valves 26 are closed, and metering is carried out through the flow meter 27, when the amount of the injection reaches the set amount, the connection pipe 25 is closed, so that the operation of controlling the culture liquid can be performed to any one of the culture tanks 21 to achieve a high-precision culture operation.
The working principle of the invention is as follows: the rotating motor 19 drives the rotating rod 7 to rotate ninety degrees clockwise, the sleeve frame plate 8 drives the linkage screw rod 9 to enable the movable screw block 10 to rotate, the camera 12 moves to the rightmost position on the upper surface of the transparent culture plate 3, the other camera 12 moves to the lower surface position of the transparent culture plate 3, the first driving motor 11 drives the linkage screw rod 9 to rotate forward, the movable screw block 10 drives the camera 12 to move backward along the upper surface of the transparent culture plate 3, the camera 12 below the transparent culture plate 3 can check the culture condition of NK cells at the bottom end of the inner wall of the culture tank 21, the second driving motor 16 drives the first belt pulley 15 to rotate forward, the first belt pulley 15 drives the movable belt 14 to transmit, the linkage block 13 drives the movable screw block 5 to enable the rotating rod 7 to drive the two sleeve frame plates 8 to move rightward, and the culture conditions of the other three culture tanks 21 are continuously checked, second driving motor 16 drives first belt pulley 15 and reverses, it resets 1 inner wall leftmost position department of incubator to cup joint framed panel 8, rotating electrical machines 19 drives dwang 7 anticlockwise rotation and resets initial position department, start moving motor 20 and drive screw 6 and rotate corotation, drive screw 6 drives removal thread piece 5 shifts up under the effect of screw thread and looks over another layer of transparent culture plate 3 can, realize the all-round effect of looking over, and realize that the accuracy is looked over, and need not open incubator 1 and can accomplish and look over the operation.
The points to be finally explained are: first, in the description of the present application, it should be noted that, unless otherwise specified and limited, the terms "mounted," "connected," and "connected" should be understood broadly, and may be a mechanical connection or an electrical connection, or a communication between two elements, and may be a direct connection, and "upper," "lower," "left," and "right" are only used to indicate a relative positional relationship, and when the absolute position of the object to be described is changed, the relative positional relationship may be changed;
secondly, the method comprises the following steps: in the drawings of the disclosed embodiments of the invention, only the structures related to the disclosed embodiments are referred to, other structures can refer to common designs, and the same embodiment and different embodiments of the invention can be combined with each other without conflict;
and finally: the present invention is not limited to the above preferred embodiments, but rather, any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. The utility model provides an in vitro isolated culture auxiliary device of NK cell convenient to operate, includes incubator (1), its characterized in that: a plurality of positioning groove plates (2) which are sequentially arranged from top to bottom are arranged in the incubator (1), a linkage support plate (4) is connected to the top ends of the positioning groove plates (2) in a sliding manner, and an inspection mechanism is arranged on one side of the linkage support plate (4);
inspection mechanism is including setting up removal screw thread piece (5) in linkage extension board (4) one side, remove inside drive screw (6) that is provided with of screw thread piece (5), dwang (7) are installed in linkage extension board (4) bottom embedding, dwang (7) outside just be located linkage extension board (4) about the equal symmetrical cover in both sides be equipped with and cup joint framed board (8), cup joint framed board (8) inner wall connection has linkage screw (9), the outside cover of linkage screw (9) is equipped with and removes spiral shell piece (10), remove spiral shell piece (10) bottom and install camera (12), linkage screw (9) one end is installed and is used for driven first driving motor (11).
2. The auxiliary device for facilitating the in vitro NK cell separation and culture of claim 1, wherein: cup joint framed lamella (8) one side and be connected with linkage block (13), linkage block (13) one side is connected with removal belt (14), remove the inside cover of belt (14) and be equipped with first belt pulley (15), second driving motor (16) are installed to first belt pulley (15) one end, second belt pulley (17) are installed to first belt pulley (15) one side, second belt pulley (17) internally mounted has rotating shaft pole (18), display (50) are installed to incubator (1) one side.
3. The auxiliary device for facilitating the in vitro NK cell separation and culture of claim 1, wherein: rotating electrical machines (19) are installed on dwang (7) top, moving electrical machines (20) are installed on drive screw (6) top, remove inside and the outside threaded connection of drive screw (6) of screw piece (5), remove inside and the outside threaded connection of linkage screw (9) of screw piece (10).
4. The auxiliary device for facilitating the in vitro NK cell separation and culture of claim 1, wherein: dwang (7) outside just is located two and cup joints position department and linkage extension board (4) inside sliding connection between framed board (8), dwang (7) outside with cup joint framed board (8) fixed connection, a plurality of culture tanks (21) have been seted up on transparent culture plate (3) top.
5. The auxiliary device for facilitating the in vitro NK cell separation and culture of claim 1, wherein: the socket frame board (8) opposite side is connected with three connecting branch (22), connecting branch (22) one end fixedly connected with cultivates the mechanism, cultivate the mechanism including setting up injection pipe (23) at connecting branch (22) one end fixed connection, injection pipe (23) outer wall below is installed three atomizing injector head (24) and outer wall top intercommunication and is had connecting pipe (25), first electric control valve (26) are installed on connecting pipe (25) top, flowmeter (27) are installed on first electric control valve (26) top, flowmeter (27) one end is connected with first shunt tubes (28), first shunt tubes (28) outer wall top intercommunication has connecting hose (29) that are used for tensile, connecting hose (29) top is connected with injection tube (30), injection pump machine (31) of carrying the culture solution is installed to injection tube (30) bottom, and the input end of the injection pump (31) is communicated with a second shunt pipe (32).
6. The auxiliary device for facilitating the in vitro NK cell separation and culture of claim 5, wherein: a second electric control valve (33) is installed at one end of the second shunt pipe (32), a first container (34) is installed at one end of the second electric control valve (33), a third electric control valve (35) is installed at the other end of the second shunt pipe (32), and a second container (36) is installed at the bottom end of the third electric control valve (35).
7. The auxiliary device for facilitating the in vitro NK cell separation and culture of claim 5, wherein: the three atomizing spray heads (24) are communicated with a connecting pipe (25), and two ends of the connecting hose (29) are respectively communicated with the first shunt pipe (28) and the injection pipe (30) in pairs.
8. The auxiliary device for facilitating the in vitro NK cell separation and culture of claim 4 is characterized in that: two hot air diffusion pipes (37) are arranged above the culture tank (21), the top ends of the hot air diffusion pipes (37) are connected with a third shunt pipe (38), the top end of the third shunt pipe (38) is communicated with an input pipe (39), one end of the input pipe (39) is provided with a shunt injection pipe (40), one side of the outer wall of the shunt injection pipe (40) is communicated with a pressure increasing pipe (41), the bottom end of the pressure increasing pipe (41) is provided with a circulating fan (42), the input end of the circulating fan (42) is communicated with a circulating box (43), a cooling water pipe (44) is arranged in the circulating box (43), one end of the cooling water pipe (44) is provided with a circulating outlet pipe (45) and the other end is provided with a circulating inlet pipe (46), and a suction pipe (47) is arranged above the circulating water inlet pipe (46), and one end of the suction pipe (47) is connected with a diffusion low suction pipe (48).
9. The auxiliary device for facilitating the in vitro NK cell separation and culture of claim 8, wherein: an electric heating plate (49) is installed at the bottom end of the circulating box (43), and the suction pipe (47) is communicated with the diffusion low suction pipe (48).
10. The auxiliary device for conveniently operating the NK cell in-vitro separation and culture is characterized by further comprising a using method for conveniently operating the NK cell in-vitro separation and culture auxiliary device according to any one of claims 1 to 9, wherein the using method comprises the following specific steps:
step one, during temperature circulation, putting NK cell culture raw materials into a culture groove (21) on each transparent culture plate (3), then aligning the transparent culture plates (3) to two positioning groove plates (2) for sliding insertion, closing a sealing door of an incubator (1), heating air inside a circulation box (43) by starting an electric heating plate (49) when the temperature is low, monitoring the inside of the circulation box (43) by using a temperature measuring instrument, starting a circulation fan (42) when the temperature reaches the NK cell culture temperature, enabling a diffusion low suction pipe (48) to suck the air inside the incubator (1) into a cooling water pipe (44), enabling the cooling water pipe (44) to enter the circulation box (43), enabling the air to enter a pressurizing pipe (41) along the circulation box (43), inputting the air into a shunt injection pipe (40) by the pressurizing pipe (41), and inputting the air into a plurality of input pipes (39) by the shunt injection pipe (40), is injected into a third shunt pipe (38) through an input pipe (39), enters a hot air diffusion pipe (37) through the third shunt pipe (38) to be uniformly blown downwards, forms multiple layers, is blown downwards from the top, then enters three through holes in the middle of a transparent culture plate (3) to be moved downwards, and hot air uniformly covers the upper surface of the transparent culture plate (3) to flow to the lower surface of the transparent culture plate (3) and then enters a diffusion low suction pipe (48) to be circulated into a suction pipe (47), so as to form thermal circulation, and when the temperature is hot, an electric heating plate (49) can be closed, a circulation outlet pipe (45) is connected with a return water pipe of a cooling water tower, and a circulation inlet pipe (46) is connected with a water inlet pipe of the cooling water tower, so as to cool the air in a circulation box (43), thereby playing a role of cooling, and the cooling air is circulated into the incubator (1) by the circulating fan (42), so that the temperature can be adjusted by cold and hot circulation;
step two, when checking the culture condition, starting a rotating motor (19) to drive a rotating rod (7) to rotate ninety degrees clockwise, driving two rotating rods (7) to rotate, sleeving a frame plate (8) to drive a linkage screw rod (9) to drive a movable screw block (10) to rotate, driving a camera (12) by the movable screw block (10) to move to the rightmost position of the upper surface of a transparent culture plate (3) and moving another camera (12) to the lower surface position of the transparent culture plate (3), starting a first driving motor (11) to drive the linkage screw rod (9) to rotate forwards, driving the movable screw block (10) by the linkage screw rod (9) to move backwards under the action of threads, driving the camera (12) to move backwards along the upper surface of the transparent culture plate (3) by the movable screw block (10), thus playing a role in checking and monitoring the upper surfaces of three culture tanks (21), the image input who looks over shows on display (50), and camera (12) of transparent culture plate (3) below can look over the cultivation situation of culture tank (21) inner wall bottom NK cell, penetrate image data and carry out two image display on display (50), and start second driving motor (16) and drive first belt pulley (15) corotation, first belt pulley (15) drive removal belt (14) and carry out the transmission, rotate second belt pulley (17) of axostylus axostyle (18) outside simultaneously and carry out the transmission, removal belt (14) can drive linkage block (13) and carry out the right side shifting, and linkage block (13) drive removal thread block (5) and move right, removal thread block (5) drive dwang (7) make two cup joint frame board (8) carry out the right side shifting, cup joint frame board (8) like this and drive camera (12) inside carry out the right side shifting to three culture tank (21) of opposite side position department and start first driving motor (11) again and drive on three culture tank (21) of opposite side position department The linkage screw (9) rotates reversely, the linkage screw (9) drives the movable screw block (10) to make the camera (12) move forwards, the culture conditions of the other three culture tanks (21) can be continuously checked, the camera (12) can move back and forth to check the culture conditions in the culture tanks (21) after moving to a certain distance to the right each time, after the culture tanks (21) on the transparent culture plate (3) below are checked, the second driving motor (16) is started to drive the first belt pulley (15) to rotate reversely, and the transmission is carried out until the socket frame plate (8) is reset to the leftmost position of the inner wall of the culture tank (1), then the rotating motor (19) is started to drive the rotating rod (7) to rotate anticlockwise to reset to the initial position, so that the socket frame plate (8) can be reset to the initial position, and then the movable motor (20) is started to drive the driving screw (6) to rotate forwards, the driving screw (6) drives the movable thread block (5) to move upwards under the action of the threads to check the culture tank (21) in the upper layer of the transparent culture plate (3), so that the multilayer transparent culture plate (3) can be checked repeatedly;
step three, during the culture fluid addition, during the viewing process, the connecting support rod (22) is driven by the sleeving frame plate (8) to move, the connecting support rod (22) drives the injection pipe (23) to move, so that the injection pipe (23) moves along the upper surface of the culture tank (21), and the three groups of atomizing injection heads (24) butt joint the three culture tanks (21), so that during the viewing process, if NK cell culture fluid is less, serum-free complete culture medium liquid inside the second container box (36) or autologous plasma inside the first container box (34) can be selected, when the culture fluid inside the first container box (34) is selected, the third electric control valve (35) can be closed, the second electric control valve (33) can be opened, when the control valve inside the second container box (36) is selected to be completely serum-free culture medium liquid, the second electric control valve (33) can be closed and opened, thus, two different culture liquids can be selected, when the culture liquid in the first container (34) is selected, the injection pump (31) can be started, the autologous plasma culture liquid in the first container (34) can enter the injection pipe (30), can be injected into the connecting hose (29) through the injection pipe (30), can enter the first shunt pipe (28) along the connecting hose (29), can shunt the flow meter (27) through the first shunt pipe (28), can enter the opened first electric control valve (26) through the flow meter (27), can be guided into the connecting pipe (25) through the first electric control valve (26) and then enter the injection pipe (23), and finally can be injected into the culture tank (21) through the atomizing injection head (24), so that the culture liquid can be uniformly atomized and injected at one time, when the culture liquid in one of the culture tanks (21) is less in the observation process of the camera head (12), the injection amount can be set, the other two first electric control valves (26) are closed, metering is carried out through the flow meter (27), and the connecting pipe (25) can be closed when the injection amount reaches the set amount, so that the culture solution control operation can be carried out in any one culture tank (21), and the high-precision culture operation can be realized.
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