An in vitro incubation and collection device for liver fluke excreted-secreted antigens
Patent Information
- Application Number
- CN202522544864.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-01
- Publication Date
- 2026-10-20
- Estimated Expiration
- 2035-12-01
Smart Images
Figure CN224863753U_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the technical field of in vitro incubation devices for liver flukes, and particularly relates to an in vitro incubation and collection device for liver fluke excretion-secretion antigens. Background Technology
[0002] Fasciola hepatica, belonging to the phylum Platyhelminthes, class Trematoda, subclass Digenea, family Fasciolidae, and genus Fasciola, commonly parasitizes the liver and bile ducts of ruminants such as cattle and sheep, causing liver fluke disease. Humans can also be infected, making it a very important zoonotic disease. The worm is flat and leaf-shaped, 20–30 mm long and 8–13 mm wide. Fasciola hepatica can cause acute and chronic hepatitis and cholangitis in animals, accompanied by systemic poisoning and nutritional deficiencies. In cattle and sheep infected with Fasciola hepatica, not only does milk production decline significantly, but the quality of meat, milk, and other livestock products, as well as related biological products, deteriorates.
[0003] Fasciola hepatica excretory secretory antigens (FhESAs) are soluble proteins, glycoproteins, and other biomolecules released into the host through excretory organs and body surface secretions during the growth and development of the worm. FhESAs mainly include proteases, antioxidant enzymes, fatty acid-binding proteins, etc., and are the core molecules of the interaction between the worm and the host. Their immunogenicity, pathogenicity, and biological functions make them a key target for research on the prevention and control of fascioliasis.
[0004] The above-mentioned research requires the in vitro culture of adult liver flukes and the collection of their excretory and secreted antigens. The amount and composition of ES antigen secretion by liver flukes vary at different culture stages, so it is necessary to collect the excretory and secreted antigens at different stages separately. Therefore, the entire collection process is quite lengthy and may take several days or even dozens of days.
[0005] Currently, FhESAs are mostly collected manually. To collect FhESAs at different stages, the culture medium in the culture dishes needs to be collected multiple times a day at fixed times and locations, and then fresh culture medium needs to be added to continue culturing. Manual collection has many drawbacks, the most prominent of which are: 1. It requires fixed-point duty day and night, which is labor-intensive; 2. Frequent contact between the operator and the equipment makes it easy to become contaminated. Utility Model Content
[0006] To address the problems in the background art, this utility model provides an in vitro incubation and collection device for liver fluke excretion-secretion antigens. This utility model achieves automated in vitro incubation of liver flukes and automated collection of their excretion-secretion antigens, avoiding the fatigue of manual day and night duty, significantly reducing the number of times the operator comes into contact with the equipment, and effectively reducing the risk of contamination.
[0007] The technical solution provided by this utility model is: an in vitro incubation and collection device for excretory-secreted antigens of liver fluke, comprising a constant-temperature, light-proof in vitro culture chamber, a low-temperature storage chamber, and an automatic control system; the in vitro culture chamber is equipped with a culture dish for holding culture medium, the upper end of the culture dish is equipped with a culture dish lid, and a fluke incubation box is detachably installed on the culture dish lid. The lower end surface of the fluke incubation box is a mesh surface. The upper part of the culture dish lid is equipped with a lifting mechanism for lifting or lowering the culture dish lid together with the fluke incubation box. The side wall of the culture dish is equipped with a culture medium inlet tube and an excretory-secreted antigen collection tube. The culture medium inlet tube is equipped with an electrically controlled switch valve A, and the excretion and secretion antigen collection tube is equipped with an electrically controlled switch valve C. An electrically controlled turntable is located at the bottom of the cryopreservation chamber, with antigen storage cups evenly spaced around its upper end. The other end of the excretion and secretion antigen collection tube extends into the cryopreservation chamber and is positioned above the antigen storage cups. The automatic control system includes a controller, whose output is electrically connected to switch valve A, switch valve C, the lifting and lowering mechanism, and the turntable, respectively, to control the opening and closing of the valves, the lifting and lowering of the culture dish lid by the lifting and lowering mechanism, and the rotation or stopping of the turntable.
[0008] A further technical solution is as follows: The lifting mechanism includes a rotating shaft A, a drive motor A, a lifting rope, and a lifting frame. The lifting frame is fixedly connected to the petri dish lid. The rotating shaft A is horizontally positioned above the in vitro incubator and can rotate relative to the in vitro incubator. The drive motor A is connected to the rotating shaft A and is used to drive the rotation of the rotating shaft A. The lower end of the lifting rope is fixedly connected to the lifting frame, and the upper end of the lifting rope is fixedly connected to the rotating shaft A. During the rotation of the rotating shaft A, the lifting rope is wound around the rotating shaft A, thereby lifting the lifting frame and then lifting the petri dish lid along with the insect incubation box. During the culture stage, the insect incubation box is immersed in the culture medium, which permeates into the insect incubation box through the mesh. When changing the culture medium, the insect incubation box is temporarily lifted to avoid adverse impact on the insects during the culture medium change. The drive motor A is electrically connected to the output of the controller, which controls the forward and reverse rotation or stops the drive motor.
[0009] Furthermore, a telescopic guide rod is installed between the petri dish lid and the in vitro incubator. The lower end of the guide rod is fixedly connected to the petri dish lid, and the upper end of the guide rod is fixedly connected to the upper end of the in vitro incubator. Under the guidance of the telescopic guide rod, the petri dish lid is lifted and lowered straight up and down without rotation or shaking.
[0010] Furthermore: A camera mounting plate is installed at the lower end of the lifting frame, and a night vision camera is eccentrically mounted at the lower end of the camera mounting plate. A telescopic rotating shaft B is installed at the upper end of the camera mounting plate, and a drive motor B is installed above the in vitro incubator. The lower end of the telescopic rotating shaft B is fixedly connected to the camera mounting plate, and the upper end of the telescopic rotating shaft B is fixedly connected to the drive motor B. The two telescopic parts of the rotating shaft B can only extend and retract relative to each other, not rotate relative to each other. The night vision camera is electrically connected to the input terminal of the controller, and the drive motor B is electrically connected to the output terminal of the controller. There is no contact crossing between the rotating shaft B and the rotating shaft A to avoid mechanical interference between them.
[0011] Furthermore, the insect incubation box is equipped with a lid with a convex lens at the top, and the lid has ventilation holes on the edge of the convex lens.
[0012] A further technical solution is as follows: a gear A is fixedly connected to the lower end of the turntable, a gear B is meshed with the gear A, and a drive motor C is coaxially connected to the gear B. The rotation of the drive motor C drives the gear B to rotate, which in turn drives the gear A and the turntable to rotate synchronously. When the drive motor C stops rotating, the turntable stops rotating. The drive motor C is electrically connected to the output terminal of the controller.
[0013] A further technical solution is as follows: the lower end of the culture dish is a cone, and an egg discharge tube is provided at the bottom of the cone. The excretory secretion antigen collection tube is located above the cone. An electric switch valve D is provided on the egg collection tube, and the switch valve D is electrically connected to the output terminal of the controller.
[0014] Furthermore: the excretory secretion antigen collection tube and the egg discharge tube are connected by a four-way valve, the switch valve C is set on the excretory secretion antigen collection tube downstream of the four-way valve, the switch valve D is set on the egg discharge tube upstream of the four-way valve, and an electric switch valve E is also set on the egg discharge tube downstream of the four-way valve. The switch valve E is electrically connected to the output terminal of the controller.
[0015] A further technical solution is that the insect incubation box has a "multi-compartment" structure inside, with each insect located in a separate compartment.
[0016] A further technical solution is as follows: a carbon dioxide gas pipe is connected to the upper end of the in vitro incubator, and an electrically operated switch valve B is installed on the pipe; a water container is placed at the bottom of the in vitro incubator; a culture medium filtration and temporary storage device is installed upstream of the culture medium inlet pipe, the culture medium filtration and temporary storage device includes a culture medium storage tank, a vacuum pump, and a filter, the filter is connected to the culture medium storage tank, and a one-way valve is installed on the connecting pipe, the vacuum pump is connected to the culture medium storage tank, and the culture medium inlet pipe is connected to the culture medium storage tank; the switch valve B and the vacuum pump are electrically connected to the output terminal of the controller.
[0017] Compared with the prior art, the beneficial effects of this utility model are as follows:
[0018] 1. This invention achieves automated incubation of liver flukes and automated collection of liver fluke excretion-secretion antigens, eliminating the need for operators to manually change the culture medium at fixed times and locations, eliminating the need for operators to be on duty day and night, and reducing the number of times operators come into contact with the vessels, effectively reducing the risk of contact contamination.
[0019] 2. The lower end of the insect incubation box in this invention has a mesh structure, and the incubation box can be lifted and lowered. The beneficial effect of the mesh structure is that even if the insects are in the incubation box, the presence of the mesh structure does not affect the immersion of the insects in the culture medium, and the mesh structure can allow insect eggs to pass through. The beneficial effect of lifting and lowering the incubation box is that when changing the culture medium, lifting the insects avoids adverse impacts from the liquid flow on the insects, thus preventing them from affecting their vitality.
[0020] 3. The convex lens in this invention can magnify the insects in the incubation box several times, facilitating clear image capture by the night vision camera. The night vision camera can be an existing infrared night vision camera. The operator can more easily observe the survival status of the insects through the night vision camera.
[0021] 4. The night vision camera of this application is mounted off-center on the camera mounting plate, and can achieve full-angle coverage observation of the insect incubation box by off-center rotation.
[0022] 5. The incubation box of this utility model has a "multi-grid" structure inside, with each insect located in a compartment. This not only effectively avoids insects gathering and making observation inconvenient, but also prevents other live insects from being touched or harmed when removing dead insects.
[0023] 6. The bottom of the culture dish of this utility model is conical, which is more conducive to the collection of parasite eggs. The culture dish has two discharge channels, one of which is a collection channel for excretory secretion antigens and the other is a discharge channel for parasite eggs. The two channels can be separated or merged, which can provide samples for both parasite egg collection experiments and excretory-secretion antigens of liver fluke. The dual-purpose design increases its functional versatility. Attached Figure Description
[0024] Figure 1 This is a simplified structural diagram of the entire utility model.
[0025] Figure 2 This is a partial enlarged view of the insect incubation box of this utility model.
[0026] Figure 3 This is a top view of the internal structure of the insect incubation box of this utility model.
[0027] Figure 4 This is a top view of the turntable and the antigen storage cup on the turntable in this utility model.
[0028] Figure 5 This is a top view diagram showing the positional relationship of the guide rod, rotating shaft A, and rotating shaft B in this utility model.
[0029] Figure 6 This is a circuit block diagram with the controller as the core of this utility model.
[0030] In the diagram: 1. Filter; 2. One-way valve; 3. Culture medium storage tank; 4. Vacuum pump; 5. Switch valve B; 6. Carbon dioxide gas pipe; 7. Culture medium inlet pipe; 8. Switch valve A; 9. Rotating shaft A; 10. Guide rod; 11. Lifting rope; 12. Rotating shaft B; 13. Drive motor B; 14. Drive motor A; 15. In vitro incubator; 16. Temperature sensor A; 17. Culture dish; 18. Culture dish lid; 19. Parasite incubation box; 20. Excretory secretion antigen collection tube; 21. Mesh frame; 22. Egg discharge tube; 23. 24. Four-way valve; 25. Switch valve C; 26. Switch valve E; 27. Egg collection dish; 28. Water container; 29. Low-temperature storage box; 30. Temperature sensor B; 31. Antigen storage cup; 32. Drive motor C; 33. Gear B; 34. Gear A; 35. Turntable; 36. Lifting frame; 37. Camera mounting plate; 38. Night vision camera; 39. Convex lens; 40. Limiting block; 41. Grid cavity; 42. Insert plate; 43. Switch valve D; 44. Auxiliary drive shaft; 45. Gear C; 46. Gear D; 47. Mesh surface. Detailed Implementation
[0031] To make the technical problems, technical solutions, and beneficial effects of this utility model clearer, the present utility model will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present utility model and are not intended to limit the present utility model.
[0032] like Figure 1 As shown, this embodiment includes a constant-temperature, opaque in vitro incubator 15, a low-temperature storage box 28, and an automatic control system.
[0033] The in vitro culture chamber 15 needs to maintain a constant temperature of approximately 37°C (simulating the body temperature of the liver fluke host). To maintain this constant temperature, a heater is installed inside the in vitro culture chamber 15; a commercially available heater is acceptable. A temperature sensor A16 is also installed inside the chamber, electrically connected to the input of the controller. The temperature sensor A16 transmits the temperature signal inside the chamber to the controller. When the temperature inside the chamber is detected to be as low as 36.5°C, the controller sends a command to turn on the heater to raise the temperature. When the temperature inside the chamber is detected to be as high as 37.5°C, the controller sends a command to turn off the heater to prevent the temperature from rising further. The reason why the in vitro culture chamber 15 is opaque is also to simulate the parasitic environment of the liver fluke. The natural parasitic sites of the liver fluke are the body cavities of the host, such as the liver, bile ducts, and abdominal cavity. These environments are completely dark and without light stimulation.
[0034] The temperature of the cryopreservation chamber 28 is maintained at approximately 4°C. Liver fluke excretory-secreting antigens collected at different times are preserved at low temperatures within the chamber to prevent deterioration of these antigens. The chamber 28 maintains its low temperature by incorporating a commercially available refrigerator. A temperature sensor B29 is also installed within the chamber, electrically connected to the input of the controller. This sensor transmits the temperature signal from the chamber to the controller. When the temperature reaches 4.5°C, the controller activates the refrigerator to lower the temperature. When the temperature drops to 3.5°C, the controller deactivates the refrigerator to prevent further temperature decreases. The chamber 28 is also light-sensitive to avoid adverse effects from light on the collected liquid.
[0035] The controller's temperature control program for the in vitro incubator 15 and the cryopreservation box 28 is the simplest and most commonly used program in the field of automatic control, which is already mastered by those skilled in the art. The program part is not the content claimed in this application, so it will not be described in detail here. This application aims to protect the structure of the product.
[0036] like Figure 1As shown, the in vitro culture chamber 15 is equipped with a culture dish 17 for holding culture medium. A culture dish lid 18 is provided at the upper end of the culture dish 17. The culture dish lid 18 can be removed from the culture dish 17. An insect incubation box 19 is detachably installed on the culture dish lid 18. The lower end surface of the insect incubation box 19 is a mesh surface 46. The mesh surface 46 can be a fine steel wire mesh. The insects survive on the mesh surface 46. The purpose of providing the mesh surface 46 at the lower end of the insect incubation box 19 is: first, when the insect incubation box 19 is immersed in the culture medium, the culture medium can penetrate into the insect incubation box 19 through the mesh surface 46, providing a simulated nutritional environment for the insects; second, the insect eggs can fall through the mesh surface 46 to the bottom of the culture dish 17, so that this application can also be used as an insect egg collection device. Third, when changing the culture medium (first drain the culture dish 17 containing excretory-secreting antigens, and then add new culture medium to the culture dish 17), the insect body can be lifted by the mesh surface 46 to avoid the discharged liquid flow from adversely impacting the insect body and affecting its vitality.
[0037] The in vitro incubator 15 has a forward-opening door structure, which allows the culture dish 17, insect incubation box 19, etc. to be taken out after the door is opened.
[0038] like Figure 2 As shown, to enable rapid removal and placement of the insect incubation box 19 from the petri dish lid 18, this embodiment has a circular hole with a diameter larger than that of the insect incubation box 19 machined in the center of the petri dish lid 18. The upper outer diameter of the insect incubation box 19 has a step that is wider at the top and narrower at the bottom. After the insect incubation box 19 is placed into the circular hole of the petri dish lid 18 from bottom to top, the step passes through the circular hole, and then two semi-circular inserts 41 are inserted into the lower end of the step to support the insect incubation box 19. Similarly, after removing the two inserts 41, the insect incubation box 19 can be removed from the petri dish lid 18 from top to bottom.
[0039] To enable automatic lifting and lowering of the culture dish 17, a lifting and lowering mechanism is provided on the upper part of the culture dish lid 18. This mechanism is used to lift or lower the culture dish lid 18 together with the insect incubation box 19. Figure 1As shown, the lifting mechanism includes a rotating shaft A9, a drive motor A14, a lifting rope 11, and a lifting frame 35. The lifting frame 35 is fixedly connected to the petri dish lid 18. The rotating shaft A9 is horizontally positioned above the in vitro culture chamber 15 and can rotate relative to the in vitro culture chamber 15. The drive motor A14 is fixedly installed on the in vitro culture chamber 15 and connected to the rotating shaft A9. The drive motor A14 is used to drive the rotation of the rotating shaft A9. The lower end of the lifting rope 11 is fixedly connected to the lifting frame 35, and the upper end of the lifting rope 11 is fixedly connected to the rotating shaft A9. During the rotation of the rotating shaft A9, the lifting rope 11 is wound around the rotating shaft A9, thereby lifting the lifting frame 35 and then lifting the petri dish lid 18 together with the insect incubation box 19. During the cultivation stage, the insect incubation box 19 is immersed in the culture medium, which permeates into the insect incubation box 19 through the mesh surface 46. When changing the culture medium, the insect incubation box 19 is temporarily lifted. The drive motor A14 is electrically connected to the output terminal of the controller. The controller can send commands to make the drive motor rotate forward or backward a specified number of times.
[0040] To prevent the insect incubation box 19 from swaying or rotating during its ascent, this embodiment provides at least two telescopic guide rods 10 between the petri dish lid 18 and the in vitro culture box 15. The lower end of the guide rod 10 is fixedly connected to the petri dish lid 18, and the upper end of the guide rod 10 is fixedly connected to the upper end of the in vitro culture box 15. Under the guidance of the telescopic guide rods 10, the petri dish lid 18 is ensured to always move straight up and down without rotation or swaying. Therefore, the insects will not experience unnecessary vibrations in the insect incubation box 19.
[0041] To improve the visibility of the insect's survival status, this application provides a camera mounting plate 36 at the lower end of the lifting frame 35. A limiting block 39 is also provided on the lifting frame 35 to limit the lower surface of the camera mounting plate 36. Under the limiting action of the limiting block 39, the camera mounting plate 36 can maintain a fixed gap with the lifting frame 35. This gap allows the camera mounting plate 36 to rotate freely relative to the lifting frame 35. A night vision camera 37 is eccentrically mounted at the lower end of the camera mounting plate 36. The significance of the eccentrically mounted night vision camera 37 is that, by rotating the camera mounting plate 36, a full-range observation of the insect incubation box 19 can be achieved. A commercially available night vision camera 37 is required; an infrared-enabled night vision camera 37 is optional. A telescopic hinge B12 is located at the upper end of the camera mounting plate 36. A drive motor B13 is located above the in vitro incubator 15. The lower end of the telescopic hinge B12 is fixedly connected to the camera mounting plate 36, and the upper end is fixedly connected to the drive motor B13. The two telescopic parts of the hinge B12 can only extend and retract relative to each other, not rotate relative to each other. The reason for designing the hinge B12 as telescopic is that the camera mounting plate 36 also needs to be lifted along with the lifting frame 35, while the drive motor B13 remains stationary. The night vision camera 37 is electrically connected to the input terminal of the controller, transmitting the video signal to the controller. The drive motor B13 is electrically connected to the output terminal of the controller, and its operation is controlled by the controller. It should be noted that the hinge B12 and the hinge A9 do not have any contact intersection to avoid mechanical interference. (Appendix) Figure 5 This is a top view of in vitro incubator 15, with attached... Figure 5 The purpose is to clearly demonstrate the positional relationship between guide rod 10, pivot A9, and pivot B12, thereby illustrating that there is no mechanical interference between them. For example... Figure 5 As shown, an auxiliary drive shaft 43 is set next to the rotating shaft A9. The auxiliary drive shaft 43 can rotate relative to the in vitro culture box 15. Two gears C44 that rotate with the shaft are fixedly set on the auxiliary drive shaft 43. The rotating shaft A9 is divided into two ends. Each segment is equipped with a gear D45 that meshes with the gear C44. In this way, the synchronous rotation of the two segments of the rotating shaft A9 is achieved through the transmission between the gears. At this time, the suspended end of the rotating shaft A9 is connected to the upper end of the in vitro culture box 15 through the lifting component. The lifting component is fixedly connected to the in vitro culture box 15. A rotating bearing is set between the rotating shaft A9 and the lifting component, so that the rotating shaft A9 can rotate relative to the lifting component.
[0042] The culture dish 17 has a culture medium inlet tube 7 and an excretory / secretory antigen collection tube 20 on its side wall. During the incubation of the worms, the excretory / secretory antigens permeate into the culture medium through the mesh surface 46. The culture medium is collected periodically to obtain the excretory / secretory antigens of the liver fluke at different time points. An electrically controlled switch valve A8 is installed on the culture medium inlet tube 7, and an electrically controlled switch valve C24 is installed on the excretory / secretory antigen collection tube 20. Switch valves A8 and C24 are electrically connected to the output of the controller, thus avoiding manual operation. The most tedious task of changing the culture medium at fixed times and locations is managed by the controller, eliminating the need for operators to work day and night and reducing the number of times operators come into contact with the dishes, effectively reducing the risk of contact contamination.
[0043] The cryogenic storage box 28 has an electrically controlled turntable 34 at its bottom. Antigen storage cups 30 are equidistantly arranged circumferentially on the upper end of the turntable 34. The other end of the excretory secretion antigen collection tube 20 extends into the cryogenic storage box 28 and is located above the antigen storage cups 30. (See attached figures for this embodiment.) Figure 4 There are eight antigen storage cups 30. The purpose of setting up eight antigen storage cups 30 is that the excretory-secreting antigen stock solution collected at different times can be dispensed into different antigen storage cups 30, without having to take out a cup each time, so the operator does not need to be on duty day and night.
[0044] During the day, antigen storage cups 30 can be removed one by one, centrifuged to obtain the supernatant, and then used for subsequent scientific research experiments.
[0045] The insect incubation box 19 is equipped with a lid with a convex lens 38 at the top. The convex lens 38 can magnify the insects in the insect incubation box 19 several times, making it easier for the night vision camera 37 to clearly capture images. The operator can more easily observe the survival status of the insects through the night vision camera 37, which is conducive to timely detection and removal of dead insects and reduces contamination of the culture medium.
[0046] The lid of the box with the convex lens 38 has a vent hole, which is used for air exchange between the inside and outside of the insect incubation box 19.
[0047] The lower end of the turntable 34 is fixedly connected to a gear A33, which meshes with a gear B32. A drive motor C31 is coaxially connected to gear B32. The rotation of the drive motor C31 drives the gear B32 to rotate, which in turn drives the gear A33 and the turntable 34 to rotate synchronously. When the drive motor C31 stops rotating, the turntable 34 stops rotating. The drive motor C31 is electrically connected to the output of the controller. The number of rotations of the drive motor C31 is set by the controller. The number of rotations corresponds to the rotation angle of the turntable 34. Each rotation of the turntable 34 corresponds to one antigen storage cup 30 and one excretory secretion antigen collection tube 20. For example, if there are eight antigen storage cups 30, the number of rotations of the turntable 34 is 1 / 8 of a rotation, and the corresponding rotation angle of the turntable 34 is 45°. The number of rotations required for the drive motor C31 to rotate once is calculated based on the transmission ratio of gear A33 and gear B32.
[0048] The lower end of the culture dish 17 is conical, which is more conducive to the accumulation of parasite eggs. An egg discharge tube 22 is located at the bottom of the conical surface, and the excretory-secretory antigen collection tube 20 is located above the conical surface. This application is mainly used to collect the excretory-secretory antigens of *Fasciola hepatica*, collected through the excretory-secretory antigen collection tube 20. If it is also necessary to collect parasite eggs, the eggs at the bottom of the culture dish 17 can be collected through the egg discharge tube 22 after the excretory-secretory antigen collection tube 20 has finished collecting. Therefore, this application has at least two functions. An electrically operated switch valve D42 is installed on the egg collection tube. The switch valve D42 is electrically connected to the output of the controller, and the opening and closing of the switch valve D42 is also achieved through automatic control.
[0049] Furthermore, such as Figure 1 As shown, the excretory-secretory antigen collection tube 20 and the egg discharge tube 22 are connected by a four-way valve 23. The switching valve C24 is located on the excretory-secretory antigen collection tube 20 downstream of the four-way valve 23, and the switching valve D42 is located on the egg discharge tube 22 upstream of the four-way valve 23. An electrically operated switching valve E25 is also located on the egg discharge tube 22 downstream of the four-way valve 23, and the switching valve E25 is electrically connected to the output of the controller. The culture dish 17 of this application has two discharge channels, one for the excretory-secretory antigen collection tube 20 and the other for the egg discharge tube 22. These two channels can be separated or merged, providing samples for both egg collection experiments and liver fluke excretory-secretory antigen experiments, thus increasing its functional versatility.
[0050] like Figure 3 As shown, the insect incubation box 19 has a "multi-grid" structure inside, with each insect located in a separate compartment 40. This not only effectively avoids insects gathering and causing inconvenience in observation, but also prevents other live insects from being touched or harmed when removing dead insects.
[0051] In addition to the above technical solutions, this application also requires the following structures:
[0052] 1. The upper end of the in vitro incubator 15 is connected to a carbon dioxide gas pipe 6 and an electric switch valve B5 is installed on the pipe to supply carbon dioxide with a concentration of about 5% into the incubator. The purpose is to maintain the pH value of the culture system and simulate the physiological environment in vivo.
[0053] 2. A water-filled dish 27 is placed at the bottom of the incubator 15. Its function is to prevent rapid evaporation of moisture from the culture medium and to stabilize the high-humidity environment, ensuring uniform temperature and humidity distribution within the incubator and reducing condensation. Simultaneously, the high humidity environment prevents CO2 from rapidly dissipating due to dryness before contact with the culture medium. The culture dish 17 is supported by a mesh frame 21, which sits atop the water-filled dish 27.
[0054] 3. A culture medium filtration and storage device is installed upstream of the culture medium inlet tube 7. Its purpose is to filter out bacteria and microorganisms in the culture medium and minimize the adverse effects on the obtained excretion and secretion antigens.
[0055] In this embodiment, the culture medium filtration and temporary storage device includes a culture medium storage tank 3, a vacuum pump 4, and a filter 1. The filter 1 is connected to the culture medium storage tank 3, and a one-way valve 2 is installed on the connecting pipe. Only filtered culture medium can enter the culture medium storage tank 3 through the one-way valve 2. The vacuum pump 4 is connected to the culture medium storage tank 3. The process of the vacuum pump 4 evacuating the culture medium storage tank 3 introduces the culture medium into the culture medium storage tank 3. The culture medium inlet pipe 7 is connected to the culture medium storage tank 3, and the culture medium is supplied to the culture dish 17 through the culture medium inlet pipe 7. The culture medium storage tank 3 can be a 37°C constant temperature chamber. The switching valve B5 and the vacuum pump 4 are electrically connected to the output terminal of the controller. The switching valve B5 and the vacuum pump 4 are connected to the automatic control system to achieve fully automatic control.
[0056] It should be noted that the programming of the controller for the working sequence, rotation speed, rotation direction, and number of rotations of the drive motor, as well as the programming of the opening and closing sequence and opening and closing duration of the switching valve, are all conventional programs and are methods that have been mastered by those skilled in the art, and will not be elaborated here.
[0057] The control system also includes a display and a remote transmission device. The display and controller are interconnected, allowing users to view required data in real time and input set parameters to the controller. The remote transmission device communicates with the display to enable remote data transmission, facilitating data reception and control via mobile phones or PCs. For example, users can remotely rotate the night vision camera 37 via mobile phone to comprehensively observe the insect's survival status. All program components involved in this application are not included in the claimed content.
[0058] The process of in vitro incubation and collection of liver fluke excretion-secretion antigens in this application is as follows:
[0059] 1. Add culture medium to culture dish 17, immerse the insect incubation box 19 in the culture medium, and after the first set time is reached, lift the insect incubation box 19 and open the switch valve C24. The culture medium containing the excreted secreted antigens enters the antigen temporary storage cup 30. After collection, the antigen temporary storage cup 30 is rotated at an angle (45° in this embodiment).
[0060] 2. Close valve C24, open valve A8, add culture medium to petri dish 17 again, and repeat the above steps.
[0061] If it is also necessary to collect parasite eggs, the switch valve E25 should be opened after each step of collecting excreted and secreted antigens to collect the parasite eggs into the additionally set parasite egg collection dish 26.
[0062] In summary, this utility model is an automated device with a novel structure, ingenious design, and low cost. It effectively replaces manual labor, reduces fatigue, improves work efficiency, and lowers the risk of infection for personnel and contamination of samples, making it very suitable for widespread use in the industry.
Claims
1. A device for in vitro incubation and collection of liver fluke excretion-secretion antigens, characterized in that: The system includes a constant-temperature, opaque in vitro incubator (15), a cryopreservation box (28), and an automatic control system. The in vitro incubator (15) contains a culture dish (17) for holding culture medium. A culture dish lid (18) is located at the top of the culture dish (17). A parasite incubation box (19) is detachably mounted on the culture dish lid (18). The lower surface of the parasite incubation box (19) has a mesh structure. A lifting mechanism is located at the top of the culture dish lid (18) to lift or lower the culture dish lid (18) along with the parasite incubation box (19). A culture medium inlet tube (7) and an excretory secretion antigen collection tube (20) are located on the side wall of the culture dish (17). An electrically controlled... The switch valve A (8) and the excretion and secretion antigen collection tube (20) are equipped with an electrically controlled switch valve C (24); the bottom of the low temperature storage box (28) is equipped with an electrically controlled turntable (34), and the upper end of the turntable (34) is equipped with antigen temporary storage cups (30) at equal intervals along the circumference. The other end of the excretion and secretion antigen collection tube (20) extends into the low temperature storage box (28) and is located above the antigen temporary storage cups (30); the automatic control system includes a controller, the output end of which is electrically connected to the switch valve A (8), the switch valve C (24), the lifting and placing mechanism and the turntable (34) respectively, and is used to control the opening and closing of the valves, the lifting and placing mechanism to lift and place the petri dish lid (18), and the rotation or stopping of the turntable (34).
2. The device for in vitro incubation and collection of liver fluke excretion-secretion antigens according to claim 1, characterized in that: The lifting mechanism includes a rotating shaft A (9), a drive motor A (14), a lifting rope (11), and a lifting frame (35). The lifting frame (35) is fixedly connected to the petri dish lid (18). The rotating shaft A (9) is horizontally positioned above the in vitro incubator (15) and can rotate relative to the in vitro incubator (15). The drive motor A (14) is connected to the rotating shaft A (9) and is used to drive the rotation of the rotating shaft A (9). The lower end of the lifting rope (11) is fixedly connected to the lifting frame (35), and the upper end of the lifting rope (11) is fixedly connected to the rotating shaft A (9). (9) Fixed connection, during the rotation of the rotating shaft A (9), the lifting rope (11) is wrapped around the rotating shaft A (9), thereby lifting the lifting frame (35), and then lifting the petri dish lid (18) together with the insect incubation box (19); during the culture stage, the insect incubation box (19) is immersed in the culture medium, and the culture medium penetrates into the insect incubation box (19) through the mesh surface (46). When changing the culture medium, the insect incubation box (19) is temporarily lifted; the drive motor A (14) is electrically connected to the output end of the controller, and the controller controls the forward and reverse rotation or stop of the drive motor.
3. The device for in vitro incubation and collection of liver fluke excretion-secretion antigens according to claim 2, characterized in that: A telescopic guide rod (10) is provided between the petri dish lid (18) and the in vitro incubator (15). The lower end of the guide rod (10) is fixedly connected to the petri dish lid (18), and the upper end of the guide rod (10) is fixedly connected to the upper end of the in vitro incubator (15). Under the guidance of the telescopic guide rod (10), the petri dish lid (18) is lifted and lowered straight up and down without rotation or shaking.
4. The device for in vitro incubation and collection of liver fluke excretion-secretion antigens according to claim 2, characterized in that: A camera mounting plate (36) is set at the lower end of the lifting frame (35). A night vision camera (37) is eccentrically mounted at the lower end of the camera mounting plate (36). A telescopic rotating shaft B (12) is set at the upper end of the camera mounting plate (36). A drive motor B (13) is set above the in vitro incubator (15). The lower end of the telescopic rotating shaft B (12) is fixedly connected to the camera mounting plate (36), and the upper end of the telescopic rotating shaft B (12) is fixedly connected to the drive motor B (13). The two telescopic parts of the rotating shaft B (12) can only extend and retract relative to each other and cannot rotate relative to each other. The night vision camera (37) is electrically connected to the input end of the controller, and the drive motor B (13) is electrically connected to the output end of the controller.
5. The device for in vitro incubation and collection of liver fluke excretion-secretion antigens according to claim 4, characterized in that: The insect incubation box (19) is provided with a lid with a convex lens (38) at the top, and the lid with ventilation holes is located on the edge of the convex lens (38).
6. The device for in vitro incubation and collection of liver fluke excretion-secretion antigens according to claim 1, characterized in that: The lower end of the turntable (34) is fixedly connected to a gear A (33), and a gear B (32) is meshed with the gear A (33). A drive motor C (31) is coaxially connected to the gear B (32). The rotation of the drive motor C (31) drives the gear B (32) to rotate, which in turn drives the gear A (33) and the turntable (34) to rotate synchronously. When the drive motor C (31) stops rotating, the turntable (34) stops rotating. The drive motor C (31) is electrically connected to the output end of the controller.
7. The device for in vitro incubation and collection of liver fluke excretion-secretion antigens according to claim 1, characterized in that: The lower end of the petri dish (17) is a cone, and an egg discharge tube (22) is provided at the bottom of the cone. The excretion secretion antigen collection tube (20) is located above the cone. An electric switch valve D (42) is provided on the egg collection tube, and the switch valve D (42) is electrically connected to the output end of the controller.
8. The device for in vitro incubation and collection of liver fluke excretion-secretion antigens according to claim 7, characterized in that: The excretory secretion antigen collection tube (20) and the egg discharge tube (22) are connected by a four-way valve (23). The switch valve C (24) is located on the excretory secretion antigen collection tube (20) downstream of the four-way valve (23). The switch valve D (42) is located on the egg discharge tube (22) upstream of the four-way valve (23). An electric switch valve E (25) is also located on the egg discharge tube (22) downstream of the four-way valve (23). The switch valve E (25) is electrically connected to the output end of the controller.
9. The device for in vitro incubation and collection of liver fluke excretion-secretion antigens according to claim 1, characterized in that: The insect incubation box (19) has a "multi-grid" structure inside, with each insect located in a separate compartment (40).
10. The device for in vitro incubation and collection of liver fluke excretion-secretion antigens according to claim 1, characterized in that: The upper end of the in vitro incubator (15) is connected to a carbon dioxide gas pipe (6) and an electric switch valve B (5) is installed on the pipe; a water container (27) is placed at the bottom of the in vitro incubator (15); a culture medium filtration and temporary storage device is installed upstream of the culture medium inlet pipe (7), the culture medium filtration and temporary storage device includes a culture medium storage tank (3), a vacuum pump (4) and a filter (1), the filter (1) is connected to the culture medium storage tank (3), and a one-way valve (2) is installed on the connecting pipe, the vacuum pump (4) is connected to the culture medium storage tank (3), and the culture medium inlet pipe (7) is connected to the culture medium storage tank (3); the switch valve B (5) and the vacuum pump (4) are electrically connected to the output end of the controller respectively.