Silkworm egg automatic positioning device for microinjection system

Abstract

The invention relates to the technical field of medicine, and discloses an automatic silkworm egg positioning device for a microinjection system, which comprises a bottom plate, the top of the bottom plate is fixedly connected with a mounting plate, the rear side of the top of the mounting plate is fixedly connected with a supporting plate, and the top of the supporting plate is fixedly connected with a connecting plate. The top of the connecting plate is fixedly connected with an integrated microscope, two eyepieces are arranged on the front side of the integrated microscope, adjusting wheels are arranged on the left side and the right side of the exterior of the integrated microscope, the front side of the connecting plate is fixedly connected with a light source, and an RGB imager is installed on the front side of the bottom of the connecting plate; and a moving assembly is arranged at the top of the mounting plate, and a driving assembly is mounted in the moving assembly. In the whole process, human errors are avoided, and the position is accurately adjusted through the RGB imager, so that accurate injection can be realized, and the success rate of transgenosis is greatly improved.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technologies, and particularly to an automatic silkworm egg positioning device for a microinjection system. Background Art

[0002] Implementing transgenes through silkworm egg injection is an important biotechnological means. During the process of silkworm transgenesis, first, a vector containing the target gene needs to be constructed. This vector usually includes components such as a promoter, the target gene, and a marker gene. Then, using microinjection technology, the constructed vector is precisely injected into silkworm eggs. When injecting, special microinjection equipment is required and operated under a microscope. An extremely fine injection needle is passed through the eggshell and egg membrane, and the vector is injected near the egg cytoplasm or nucleus. Subsequently, the injected vector undergoes processes such as integration within the cells of the silkworm egg. As the silkworm egg develops, the target gene has the opportunity to integrate into the silkworm genome, enabling the silkworm to express new traits during subsequent growth and development. The marker gene helps to screen out successfully transgenic silkworm individuals, thus achieving the transgenic modification of silkworms. This provides a basis for many applications such as silkworm gene function research, variety improvement, and production of silk with special properties.

[0003] The existing implementation is to manually paste silkworm eggs onto a glass slide, fix the glass slide on the stage, and then, by controlling the manipulation position of the microinjection system, move the steel needle and the glass needle tip of the manipulation arm to the position of each silkworm egg. After the steel needle pierces the eggshell, the plasmid solution is injected into the silkworm egg using the glass needle.

[0004] However, manually pasting silkworm eggs is an extremely time-consuming task; in addition, the position matching between the tips of the glass needle and the steel needle on the manipulation arm and the silkworm eggs, as well as the needle insertion force, etc., extremely test the experience of the experimenter. These problems will all lead to a relatively low efficiency and success rate of silkworm transgenesis. Therefore, in view of the above deficiencies, an automatic silkworm egg positioning device for a microinjection system is proposed to solve the above problems. Summary of the Invention

[0005] To make up for the above deficiencies, the present invention provides an automatic silkworm egg positioning device for a microinjection system, aiming to improve the problems in the prior art that manually pasting silkworm eggs is time-consuming and laborious and the needle tip is not convenient for precisely aligning the position of the silkworm eggs.

[0006] To achieve the above objective, the present invention adopts the following technical solutions:

[0007] An automatic silkworm egg positioning device for a microinjection system, comprising a bottom plate, a mounting plate fixedly connected to the top of the bottom plate, a support plate fixedly connected to the rear side of the top of the mounting plate, a connecting plate fixedly connected to the top of the support plate, a stereomicroscope fixedly connected to the top of the connecting plate, two eyepieces arranged on the front side of the stereomicroscope, adjusting wheels arranged on both the left and right sides of the outside of the stereomicroscope, a light source fixedly connected to the front side of the connecting plate, an RGB imager installed on the front side of the bottom of the connecting plate, a moving component arranged on the top of the mounting plate, a driving component installed inside the moving component, a stage fixedly connected to the top of the driving component, a silkworm egg plate arranged inside the stage, controllers installed on both the left and right sides of the top of the bottom plate, and injection components arranged on both the left and right sides of the top of the bottom plate;

[0008] As a further description of the above technical solution:

[0009] The moving component includes a moving platform, the bottom of the moving platform is installed on the top of the mounting plate, a bearing plate is slidably connected to the top of the moving platform, and an adjusting handle is arranged on the left side of the top of the moving platform;

[0010] As a further description of the above technical solution:

[0011] The driving component includes a motor, the outside of the motor is installed inside the bearing plate, and the output end of the motor is fixedly connected to a driving shaft;

[0012] As a further description of the above technical solution:

[0013] Two sliding rods are slidably connected inside the stage, springs are sleeved on the outside of both sliding rods, fixing rings are fixedly connected to the outside of both sliding rods, pulling plates are fixedly connected to the far sides of both sliding rods, two clamping plates are fixedly connected to the near sides of both sliding rods, two limiting frames are fixedly connected to the bottom of the stage, and limiting grooves are opened inside the bearing plate;

[0014] As a further description of the above technical solution:

[0015] Both injection components include adjusting frames, the bottoms of both adjusting frames are installed on the top of the bottom plate, two steel needles are fixedly connected to the front sides of both adjusting frames, two glass needles are fixedly connected to the front sides of both adjusting frames, and a delivery pipe is detachably connected to the far side of both glass needles;

[0016] As a further description of the above technical solution:

[0017] The outside of the fixed ring is slidably connected to the inside of the stage, and the outside of the clamping plate is slidably connected to the inside of the stage;

[0018] As a further description of the above technical solution:

[0019] One end of the spring is fixedly connected to the outside of the fixed ring, and the other end of the spring is fixedly connected to the inside of the stage;

[0020] As a further description of the above technical solution:

[0021] The outside of the limit frame is slidably connected to the inside of the limit groove, and a plasmid solution injector is connected to the outside of the delivery pipe.

[0022] The present invention has the following beneficial effects:

[0023] 1. In the present invention, the stereomicroscope is responsible for obtaining a three-dimensional clear image to facilitate accurate positioning of the silkworm eggs and the injection site; the RGB imager uses advanced image acquisition technology to quickly capture the image information of the silkworm eggs. Through the analysis of the built-in intelligent algorithm, the position coordinates of the silkworm eggs are accurately determined; the controller receives the image data of the RGB imager, and after rapid processing and analysis, it issues precise control instructions to the actuators such as the motor and the moving platform. During the operation, first, the moving platform drives the carrier plate to displace, moves the silkworm eggs under the RGB imager for automatic position analysis, then starts the motor to drive the stage to rotate for precise adjustment, and finally controls the three-dimensional spatial position of the steel needle and the glass needle through the adjustment frame to pierce the eggshell and inject the plasmid solution. The whole process avoids human errors and realizes accurate injection, greatly improving the success rate of transgenesis.

[0024] 2. In the present invention, by pulling the pull plate, the sliding rod displaces and the spring compresses. After releasing the pull plate, the spring releases energy to drive the clamping plate to reset. The clamping plate closely fits the edge of the silkworm egg plate, and uses the elastic force and friction force to firmly fix the silkworm egg plate inside the stage, ensuring that the silkworm egg plate can remain stable during the entire operation process of placing the silkworm egg plate, moving, rotating the stage, and injecting, providing a solid foundation for precise operation. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 is a perspective view of a silkworm egg automatic positioning device for a microinjection system proposed by the present invention;

[0026] Figure 2 is a schematic structural diagram of a connecting plate of a silkworm egg automatic positioning device for a microinjection system proposed by the present invention;

[0027] Figure 3 is a schematic structural diagram of a carrier plate of a silkworm egg automatic positioning device for a microinjection system proposed by the present invention;

[0028] Figure 4 is Figure 3 The enlarged view of part A in

[0029] Figure 5 Schematic diagram of the steel needle structure of an automatic silkworm egg positioning device for a microinjection system proposed by the present invention.

[0030] Marking description:

[0031] 1. Bottom plate; 2. Mounting plate; 3. Support plate; 4. Connecting plate; 5. Stereomicroscope; 6. Eyepiece; 7. Adjusting wheel; 8. Light source; 9. RGB imager; 10. Moving platform; 11. Carrier plate; 12. Motor; 13. Driving shaft; 14. Stage; 15. Slide bar; 16. Spring; 17. Fixed ring; 18. Pulling plate; 19. Limiting frame; 20. Limiting groove; 21. Adjusting handle; 22. Silkworm egg plate; 23. Controller; 24. Adjusting frame; 25. Steel needle; 26. Glass needle; 27. Delivery pipe; 28. Clamping plate. Specific implementation manner

[0032] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0033] Referring to Figures 1 to 3 , an embodiment provided by the present invention: An automatic silkworm egg positioning device for a microinjection system includes a bottom plate 1. The bottom plate 1 serves as the basic support component of the entire device, providing a stable bearing plane. The top of the bottom plate 1 is fixedly connected with a mounting plate 2. The rear side of the top of the mounting plate 2 is fixedly connected with a support plate 3. The top of the support plate 3 is fixedly connected with a connecting plate 4. The top of the connecting plate 4 is fixedly connected with a stereomicroscope 5. The stereomicroscope 5 is the core optical instrument for observing the fine structure of silkworm eggs and the injection process. The support plate 3 and the connecting plate 4 provide a solid vertical support force. At the same time, the height of the support plate 3 needs to be designed to adapt to the observation angle requirements of the stereomicroscope 5 to ensure that the microscope can be at an appropriate working height, facilitating the operator to observe and operate the silkworm eggs. Two eyepieces 6 are arranged on the front side of the stereomicroscope 5. The eyepieces 6 allow the operator to observe with both eyes simultaneously, obtaining a clear three-dimensional image, which is convenient for accurately positioning the silkworm eggs and the injection site. Adjusting wheels 7 are arranged on both the left and right sides of the outside of the stereomicroscope 5. The adjusting wheels 7 facilitate the operator to flexibly adjust parameters such as the focal length and magnification of the microscope according to actual needs. A light source 8 is fixedly connected to the front side of the connecting plate 4. The light source 8 provides sufficient and uniform illumination light for the observation area of the stereomicroscope 5;

[0034] The front side of the bottom of the connecting plate 4 is equipped with an RGB imager 9. Using advanced image acquisition technology, the RGB imager 9 can quickly and automatically capture the image information of the silkworm eggs on the silkworm egg plate 22, and analyze the image through the built-in intelligent algorithm to accurately determine the position coordinates of the silkworm eggs. A moving component is arranged at the top of the mounting plate 2. The moving component includes a moving platform 10, and the moving platform 10 can drive the bearing plate 11 to perform high-precision displacement movement on the horizontal plane. The bottom of the moving platform 10 is mounted on the top of the mounting plate 2, and the top of the moving platform 10 is slidably connected with a bearing plate 11. A regulating handle 21 is arranged on the left side of the top of the moving platform 10. A driving component is installed inside the moving component. The driving component includes a motor 12, and the outside of the motor 12 is installed inside the bearing plate 11. The motor 12 serves as the power source for driving the carrier table 14 to rotate. The output end of the motor 12 is fixedly connected with a driving shaft 13. The top of the driving component is fixedly connected with a carrier table 14, and the carrier table 14 is used for carrying the silkworm egg plate 22. A silkworm egg plate 22 is arranged inside the carrier table 14. On the left and right sides of the top of the bottom plate 1, controllers 23 are installed. On the one hand, the controllers 23 receive the image data from the RGB imager 9, perform rapid processing and analysis, and issue precise control instructions to the actuators such as the motor 12 and the moving platform 10 according to the preset program logic. On the other hand, the operator can also adjust the various parameters of the device through the human-machine interaction interface on the controllers 23. Injection components are arranged on the left and right sides of the top of the bottom plate 1.

[0035] Refer to Figures 2 to 4 , two sliding rods 15 are slidably connected inside the carrier table 14. When the pull plate 18 is pulled, the sliding rods 15 can smoothly displace along the slideway inside the carrier table 14. Springs 16 are sleeved outside both of the two sliding rods 15. Fixed rings 17 are fixedly connected to the outside of both of the two sliding rods 15. The outside of the fixed rings 17 is slidably connected inside the carrier table 14. One end of the spring 16 is fixedly connected to the outside of the fixed ring 17, and the other end of the spring 16 is fixedly connected to the inside of the carrier table 14. Elastic potential energy is stored when the pull plate 18 is pulled, and the energy is released after the pull plate 18 is released, driving the clamping plate 28 to reset and realizing the automatic clamping of the silkworm egg plate 22. Pull plates 18 are fixedly connected to the far sides of the two sliding rods 15, and two clamping plates 28 are fixedly connected to the near sides of the two sliding rods 15. When the spring 16 resets, the clamping plates 28 can closely fit the edge of the silkworm egg plate 22, and the silkworm egg plate 22 is firmly fixed inside the carrier table 14 by using the elastic force and friction force to ensure the stability of the silkworm egg plate 22 during the whole operation process. The outside of the clamping plate 28 is slidably connected inside the carrier table 14. Two limiting frames 19 are fixedly connected to the bottom of the carrier table 14, and limiting grooves 20 are opened inside the bearing plate 11. The outside of the limiting frames 19 is slidably connected inside the limiting grooves 20.

[0036] Refer to Figure 1 andFigure 5 , both injection components include an adjustment frame 24. The adjustment frame 24 serves as a support and adjustment structure for the steel needle 25 and the glass needle 26, and has the adjustment function of multiple degrees of freedom. The operator can precisely control the positions of the steel needle 25 and the glass needle 26 in the three-dimensional space through the adjustment frame 24, so that they are accurately aligned with the injection site of the silkworm eggs. The bottoms of the two adjustment frames 24 are both installed on the top of the bottom plate 1. Two steel needles 25 are fixedly connected to the front sides of the two adjustment frames 24. The steel needle 25 can accurately pierce the eggshell of the silkworm eggs without damaging the internal tissues of the silkworm eggs, opening up a channel for the subsequent injection of the plasmid solution by the glass needle 26. Two glass needles 26 are fixedly connected to the front sides of the two adjustment frames 24. The glass needle 26 is hollow inside and is used to transport the plasmid solution. After the steel needle 25 pierces the eggshell, it can accurately inject the plasmid solution into the silkworm eggs. A delivery tube 27 is detachably connected to the far side of the two glass needles 26, and an injection instrument for providing the plasmid solution is connected to the outside of the delivery tube 27.

[0037] Working principle: When injecting the plasmid solution into the silkworm eggs, first, the pull plate 18 can be pulled to drive the sliding rod 15 to displace, and then the fixed ring 17 can slide inside the carrier table 14. At this time, the spring 16 will be compressed, and then the clamping plate 28 will be driven to displace. At this time, the silkworm egg plate 22 can be placed inside the carrier table 14. Then, the pull plate 18 can be released, and the clamping plate 28 can be reset by the elastic force of the spring 16, so as to fix the silkworm egg plate 22. Then, the moving platform 10 can be started to drive the bearing plate 11 to displace, and then the silkworm eggs on the top of the silkworm egg plate 22 can be moved below the RGB imager 9. Then, the position of the silkworm eggs can be automatically analyzed by the RGB imager 9. Then, the motor 12 can be started to drive the bearing plate 11 to rotate, and then the carrier table 14 can be driven to rotate. At this time, the limit frame 19 will slide inside the limit groove 20, so as to accurately adjust the position of the silkworm eggs. Then, the steel needle 25 can be driven to displace by the adjustment frame 24 to pierce the eggshell of the silkworm eggs. Finally, the plasmid solution can be injected into the silkworm eggs by the glass needle 26, so as to avoid human errors, achieve accurate injection, and improve the success rate of transgenesis.

[0038] Finally, it should be noted that the above are only the preferred embodiments of the present invention and are not used to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, for those skilled in the art, they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An automatic silkworm egg positioning device for a microinjection system, comprising a bottom plate (1), characterized in that: The top of the bottom plate (1) is fixedly connected to a mounting plate (2), the top rear side of the mounting plate (2) is fixedly connected to a support plate (3), the top of the support plate (3) is fixedly connected to a connecting plate (4), the top of the connecting plate (4) is fixedly connected to a stereo microscope (5), the front side of the stereo microscope (5) is provided with two eyepieces (6), the left and right sides of the outside of the stereo microscope (5) are provided with adjustment wheels (7), the front side of the connecting plate (4) is fixedly connected to a light source (8), the bottom front side of the connecting plate (4) is installed with an RGB imager (9), the top of the mounting plate (2) is provided with a moving component, the inside of the moving component is installed with a driving component, the top of the driving component is fixedly connected to a stage (14), the inside of the stage (14) is provided with a silkworm egg plate (22), the left and right sides of the top of the bottom plate (1) are installed with controllers (23), and the left and right sides of the top of the bottom plate (1) are provided with injection components.

2. The automatic silkworm egg positioning device for a microinjection system according to claim 1, characterized in that: The mobile assembly comprises a mobile platform (10), the bottom of the mobile platform (10) is mounted on the top of the mounting plate (2), the top of the mobile platform (10) is slidably connected to a bearing plate (11), and an adjustment handle (21) is provided on the left side of the top of the mobile platform (10).

3. The automatic silkworm egg positioning device for a microinjection system according to claim 2, characterized in that: The driving assembly comprises a motor (12), the exterior of the motor (12) being mounted inside the bearing plate (11), and the output end of the motor (12) being fixedly connected to a driving shaft (13).

4. The automatic silkworm egg positioning device for a microinjection system according to claim 3, characterized in that: The loading platform (14) is internally slidably connected to two sliding rods (15), the exteriors of the two sliding rods (15) are sleeved with springs (16), the exteriors of the two sliding rods (15) are fixedly connected to fixing rings (17), the far sides of the two sliding rods (15) are fixedly connected to pull plates (18), the near sides of the two sliding rods (15) are fixedly connected to two clamping plates (28), the bottom of the loading platform (14) is fixedly connected to two limit frames (19), and the interior of the carrying plate (11) is provided with a limit groove (20).

5. The automatic silkworm egg positioning device for a microinjection system according to claim 4, characterized in that: The two injection assemblies each comprise an adjustment frame (24), the bottoms of the two adjustment frames (24) being mounted on the top of the bottom plate (1), the front sides of the two adjustment frames (24) being fixedly connected to two steel needles (25), the front sides of the two adjustment frames (24) being fixedly connected to two glass needles (26), and the far sides of the two glass needles (26) being detachably connected to a delivery tube (27).

6. The automatic silkworm egg positioning device for a microinjection system according to claim 4, characterized in that: The outside of the fixing ring (17) is slidably connected to the inside of the loading platform (14), and the outside of the clamping plate (28) is slidably connected to the inside of the loading platform (14).

7. The automatic silkworm egg positioning device for a microinjection system according to claim 4, characterized in that: One end of the spring (16) is fixedly connected to the outside of the fixing ring (17), and the other end of the spring (16) is fixedly connected to the inside of the loading platform (14).

8. The automatic silkworm egg positioning device for a microinjection system according to claim 5, characterized in that: The outside of the limiting frame (19) is slidably connected to the inside of the limiting groove (20), and the outside of the delivery tube (27) is connected to an injection device for providing a plasmid solution.

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