A semi-mechanical device for flushing embryos from oviducts in mice

Abstract

The application discloses a kind of mouse in vivo fertilization oviduct semi-mechanical embryo flushing device, about the technical field of experimental animal equipment, including embryo flushing dish and embryo flushing device, the embryo flushing dish is barrel-shaped, the embryo flushing dish is covered with embryo flushing cover above;Increase the movable oviduct placement dish, can be installed detachable, convenient and practical, avoid the waste of embryo when taking out oviduct.The embryo flushing dish increases the constant-temperature heating ring connected with the embryo flushing dish, reduces the influence of embryo by temperature, ensures the quality of embryo;Embryo flushing needle can realize mouse in vivo fertilization oviduct semi-mechanical, with a large number of water storage, quantitative liquid flushing, self-constant-temperature heating and other advantages;Optimize traditional operation, reduce the workload of personnel, while improving the quantity of embryo collection and experimental yield.

Description

Technical Field

[0001] This invention relates to the technical field of equipment for laboratory animals, and in particular to a semi-mechanized embryo flushing device for in vivo fertilization of oviducts in rodents. Background Technology

[0002] In vivo fertilization in laboratory mice is less affected by external factors than in vitro fertilization. Rare male mice can be reused without euthanasia, and the resulting embryos are of high quality with fewer abnormal cells and a high implantation and development rate.

[0003] In vivo fertilization (IVF) requires embryo flushing, a procedure where a flushing needle is inserted into the fimbriae of the fallopian tubes of mice and rats. The solution in the syringe is then pushed to flush the embryos completely out of the uterus and fallopian tubes, facilitating in vitro embryo collection or transfer. Traditional flushing procedures present several challenges. Improper handling by the personnel collecting the fallopian tubes can make locating the fimbriae under a microscope more difficult. The needle tip is quite sharp, and improper handling can puncture the fallopian tubes, damaging their integrity and causing leakage of fluid and air, preventing complete embryo flushing. Traditional flushing needles are typically about the size of a 1mL syringe, with a small handle that is inconvenient to hold and lacks protective measures for the needle tip and the person. Multiple fluid replenishments are usually required during flushing to maintain fluid levels and temperature. Embryos are sensitive to temperature changes; rapid cooling of the soaking solution can affect embryo quality and reduce transfer rates. The force and speed of flushing also affect embryo morphology and quality; excessive force can damage the embryo. Deformation, and in severe cases, damage or even detachment of the zona pellucida; the fimbriae of the fallopian tube are generally everted and relatively lubricated, and the insertion depth, fit, and seal of the needle tip when inserting into the fallopian tube will affect the difficulty of embryo flushing; poor sealing at the connection between the syringe and the plunger can also lead to leakage of the flushing fluid or the entry of bacteria; the flushing fluid is immersed in the same droplet as organs such as the fallopian tube, and since the organs are large, a lot of flushing fluid and embryos will be taken away when the organs are removed, resulting in the loss and waste of embryos; during the operation, embryos need to be collected once every 1-2 mice, and if the time is too long, the droplets will evaporate, the microscope will get hot from prolonged operation, or the use of a microscope without heating will cause the droplets to cool down, and long-term exposure to the fluid will increase the possibility of cell contamination.

[0004] Under a microscope, the approximate location of cells can be seen in the mouse oviduct, but the smaller size of the mouse oviduct increases the difficulty of embryo flushing; although the fimbriae of the rat oviduct have large openings, the thicker tube walls make it impossible to observe the location of cells under a microscope. In addition, rat embryos are more sensitive to external changes, requiring stricter operation time and techniques.

[0005] The information disclosed in this background section is intended only to enhance the understanding of the overall background of the invention and should not be construed as an admission or in any way implying that the information constitutes prior art known to those skilled in the art. Summary of the Invention

[0006] The purpose of this invention is to provide a semi-mechanized embryo flushing device for in vivo fertilization of rodent oviducts, which can solve the problem of complex operation requiring multiple fluid replenishments to ensure the liquid temperature during the embryo flushing process; and solve the problem of poor sealing at the connection between the syringe and the needle plug caused by the outward turning and relatively lubricated state of the fimbriae of the oviduct.

[0007] To achieve the above objectives, the present invention provides a semi-mechanized embryo flushing device for in vivo fertilization of rodent oviducts, comprising an embryo flushing dish and an embryo flushing apparatus, wherein the embryo flushing dish is cylindrical and covered with an embryo flushing cover.

[0008] A movable fallopian tube placement dish is placed at the bottom of the embryo flushing dish. The fallopian tube is placed at the bottom of the movable fallopian tube placement dish. The movable fallopian tube placement dish includes an upper edge and a lower edge. The upper edge and the lower edge are connected by multiple connecting skeletons. A hollow groove is formed between the upper edge, the lower edge and the multiple connecting skeletons. Multiple support platforms are provided around the bottom of the movable fallopian tube placement dish, which can suspend and support the movable fallopian tube placement dish in the embryo flushing dish.

[0009] The embryo flushing device includes an embryo flushing needle, a water collection tank, and a push handle. The embryo flushing needle and the water collection tank are connected through the embryo flushing needle interface. The embryo flushing needle is covered by an embryo flushing handle, one end of which is connected to the outer surface of the water collection tank. The needle tip of the embryo flushing needle extends beyond the other end of the embryo flushing handle. A soft plastic, adjustable-size fallopian tube fixing ring is fitted around one end of the embryo flushing handle near the embryo flushing needle. The inner ring of the fallopian tube fixing ring has fixing teeth for fixing the fixing ring to the outer wall of the fallopian tube port. The end of the water collection tank away from the embryo flushing needle is filled with a movable handle. The side of the movable handle away from the water collection tank is connected to the push handle. The end of the push handle away from the water collection tank has a push buckle. An operating ring is connected to the push handle near the push buckle. A negative pressure spring is fitted around the push handle between the movable handle and the operating ring.

[0010] Preferably, the movable fallopian tube placement dish is cylindrical.

[0011] Furthermore, a heating coil is connected to the outside of the blanking pan. The heating coil is sleeved around the lower surface of the outer periphery of the blanking pan. The heating coil includes a heating coil body, a wire, and a plug. The heater is a detachable external heater.

[0012] Preferably, the tooth profile of the fixing clamp is an obtuse triangle.

[0013] Furthermore, the blank punch also includes a fixed ring size adjustment line and a fixed ring length adjustment buckle. The fixed clamping ring is connected to the fixed ring length adjustment buckle through the fixed ring size adjustment line. The fixed ring length adjustment buckle is fixed on the blank punch handle. The fixed ring length adjustment buckle is provided with fixed ring length adjustment teeth, which can extend or contract the fixed ring size adjustment line to wrap around the fixed ring length adjustment teeth to adjust the length and width of the fixed ring size adjustment line, thereby adjusting the size of the fixed clamping ring.

[0014] Furthermore, the blank punch also includes a fixed ring length adjustment tooth and a fixed ring length adjustment line, and the operating ring is provided with a fixed ring length adjustment wheel; one end of the fixed ring length adjustment line is connected to the fixed ring length adjustment buckle and extends to the operating ring at the top of the push handle, and the other end of the fixed ring length adjustment line is connected to the fixed ring length adjustment wheel. Rotating the fixed ring length adjustment wheel can change the length of the fixed ring length adjustment line, so that the fixed clamping ring can move along the blank punch handle with the fixed ring length adjustment buckle, thereby adjusting the distance between the fixed clamping ring and the blank punch needle head.

[0015] Preferably, the water collection tank is provided with a button battery slot for placing button batteries, the outer wall of the water collection tank is provided with scale lines to indicate the liquid content in the water collection tank, the water collection tank is covered with a heating plate to maintain the liquid in the water collection tank at the required temperature, and the surface of the water collection tank is provided with a water inlet and a negative pressure vent to maintain the negative pressure state in the water collection tank.

[0016] Compared with the prior art, the semi-mechanized oviduct flushing device for in vivo fertilization in rodents according to the present invention has the following advantages:

[0017] 1. In cases such as autumn and winter, sudden temperature drops, or when the microscope does not have a bottom heating function, a heating coil is connected to the outside of the embryo flushing dish during the embryo flushing process to maintain a constant temperature. When not in use, the heater can be removed and placed separately to reduce the impact of temperature on the embryo and ensure the quality of the embryo.

[0018] 2. A movable fallopian tube placement dish has been added, which can fix the fallopian tube. It can be installed and removed, which is convenient and practical and avoids the waste of embryos when removing the fallopian tube. The movable fallopian tube placement dish includes a hollow groove. During embryo flushing, the fluid mixed with the embryo can flow into the flushing dish through the hollow groove around the perimeter, thereby collecting the flushing fluid. Multiple support platforms are provided around the bottom of the movable fallopian tube placement dish, which can suspend and support the movable fallopian tube placement dish in the middle of the bottom of the flushing dish.

[0019] 3. A negative pressure spring is fitted on the long handle. After the long handle is pushed to move the handle, the negative pressure spring can cause the long handle to spring back to its original position, thereby achieving quantitative flushing. The water collection tank is covered with a heating plate, which can maintain the liquid temperature at 37℃ or other required temperatures. The surface of the water collection tank is equipped with a water inlet and a negative pressure vent to maintain the negative pressure state inside the water collection tank, which can realize large-scale water storage and automatic water replenishment.

[0020] 4. By adjusting the size adjustment line and length adjustment line of the fixing ring, the size and length of the fixing ring and its position on the fallopian tube can be adjusted, so that the end of the embryo flushing handle and the opening of the fallopian tube fit tightly, reducing the workload of personnel and increasing the number of embryos collected and the output of the experiment. Attached Figure Description

[0021] Figure 1 This is a schematic diagram of the structure of a mold-making vessel according to an embodiment of the present invention;

[0022] Figure 2 This is a schematic diagram of the upper surface structure of a blanking vessel according to an embodiment of the present invention;

[0023] Figure 3 This is a schematic diagram of a movable fallopian tube placement dish according to an embodiment of the present invention;

[0024] Figure 4 This is a schematic diagram of the structure of an external constant temperature heating coil for a blank-filling dish according to an embodiment of the present invention;

[0025] Figure 5 This is a schematic diagram of the structure of a blank punch according to an embodiment of the present invention;

[0026] Figure 6 This is a schematic diagram of the structure of a water collection tank according to an embodiment of the present invention.

[0027] Explanation of key figure labels:

[0028] 1: Embryo flushing dish, 2: Embryo flushing cover, 3: Movable fallopian tube placement dish, 4: Connecting skeleton, 5: Hollowed-out groove, 6: Support platform, 7: External constant temperature heating coil, 8: Wire, 9: Plug, 10: Embryo flushing needle, 11: Fixing clamp ring, 12: Embryo flushing long handle, 13: Fixing clamp teeth, 14: Fixing ring size adjustment line, 15: Fixing ring length adjustment buckle, 16: Fixing ring length adjustment teeth, 17: Embryo flushing needle interface, 18: Button battery slot, 19: Button battery, 20: Heating plate, 21: Scale line, 22: Water inlet, 23: Negative pressure outlet, 24: Moving handle, 25: Negative pressure spring, 26: Pushing long handle, 27: Operating ring, 28: Pushing buckle, 29: Fixing ring length adjustment line, 30: Fixing ring length adjustment wheel, 31: Water collection tank suspension buckle. Detailed Implementation

[0029] The specific embodiments of the present invention will now be described in detail with reference to the accompanying drawings, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments.

[0030] Unless otherwise expressly stated, throughout the specification and claims, the term "comprising" or its variations such as "including" or "comprises" shall be understood to include the stated elements or components without excluding other elements or other components.

[0031] According to a preferred embodiment of the present invention, a semi-mechanized embryo flushing device for in vivo fertilization of mice is provided. The device can achieve large-volume water storage, quantitative flushing, automatic water replenishment, and constant temperature heating. At the same time, it can fix the fallopian tube, collect a large amount of flushing fluid, and retain cells. It can optimize the loss and damage of embryos in the traditional embryo flushing process and increase the number of embryos collected and the yield of experiments.

[0032] A semi-mechanized oviduct flushing device for in vivo fertilization in rodents, comprising an oviduct flushing dish 1 and an oviduct flushing apparatus.

[0033] like Figures 1 to 4 As shown, the embryo flushing dish 1 is cylindrical, and an embryo flushing cover 2 covers the top of the embryo flushing dish 1. A movable fallopian tube placement dish 3 is placed at the bottom of the embryo flushing dish 1. The movable fallopian tube placement dish 3 can be placed at the center of the bottom of the embryo flushing dish 1. The bottom of the movable fallopian tube placement dish 3 holds the fallopian tube. The movable fallopian tube placement dish 3 is cylindrical, and its diameter is smaller than that of the embryo flushing dish 1. The movable fallopian tube placement dish 3 includes an upper edge and a lower edge, which are connected by multiple connecting skeletons 4. A hollow groove 5 is formed between the upper edge, the lower edge, and the multiple connecting skeletons 4. During embryo flushing, the liquid mixed with the embryo can flow into the embryo flushing dish 1 through the hollow groove 5 around the perimeter, thereby collecting the embryo flushing fluid. Multiple support platforms 6 are provided around the bottom perimeter of the movable fallopian tube placement dish 3, which can suspend and support the movable fallopian tube placement dish 3 at the center of the bottom of the embryo flushing dish 1.

[0034] In cases such as autumn and winter, sudden temperature drops, or when the microscope does not have a bottom heating function, a heating coil is connected to the outside of the embryo-filling dish 1 during the embryo-filling process. The heating coil is an external constant temperature heating coil 7. The heating coil can be wrapped around the lower 1 / 3 of the outer circumference of the embryo-filling dish 1. The heating coil includes the heating coil body, the wire 8, and the plug 9. The embryo-filling dish 1 can be heated by plugging in the power. When not in use, the heater can be removed and placed separately.

[0035] The semi-mechanized oviduct flushing device for in vivo fertilization in rodents features a movable oviduct placement dish 3, which is easy to install and remove, preventing embryo waste during oviduct removal. The flushing dish 1 is equipped with an external constant-temperature heating coil 7 to reduce temperature-related impacts on the embryo and ensure embryo quality.

[0036] like Figure 5 and Figure 6 As shown, the blank punch includes a blank punching needle 10, a fixing clamping ring 11, a fixing ring size adjustment line 14, a fixing ring length adjustment buckle 15, a blank punching handle 12, a water collection tank, a pushing handle 26, an operating ring 27, and a pushing buckle 28.

[0037] The embryo flushing needle 10 and the water collection tank are connected through the embryo flushing needle interface 17. The embryo flushing needle 10 is covered with an embryo flushing long handle 12 to protect the structure of the embryo flushing needle 10 from bending. One end of the embryo flushing long handle 12 is connected to the outer surface of the water collection tank, and the needle length of the embryo flushing needle 10 extends beyond the end of the embryo flushing long handle 12. At the end of the embryo flushing long handle 12, a soft plastic adjustable fallopian tube fixing clip 11 is fitted. The inner ring of the fallopian tube fixing clip 11 is provided with obtuse-angled triangular fixing teeth 13 for fixing the fixing clip 11 to the outer wall of the fallopian tube port. The fixed clamping ring 11 is connected to the fixed ring length adjusting buckle 15 via the fixed ring size adjustment line 14. The fixed ring length adjusting buckle 15 is looped around the middle of the long shank of the blank 12. The fixed ring length adjusting buckle 15 is provided with fixed ring length adjusting teeth 16, which can extend or contract the fixed ring size adjustment line 14 and wrap it around the fixed ring length adjusting teeth 16 to adjust the length and width of the fixed ring size adjustment line 14, thereby adjusting the size of the fixed clamping ring 11.

[0038] One end of the water collection tank is connected to the flushing needle 10, and the other end is filled by the moving handle 23. A push handle 26 is connected to the side of the moving handle 23 away from the water collection tank. A push buckle 28 is provided on the end of the push handle 26 away from the water collection tank for manual pushing and pulling. An operating ring 27 is connected to the side of the push handle 26 near the push buckle 28. The operating ring 27 has a fixed ring length adjustment wheel 30. A negative pressure spring 25 is sleeved on the push handle 26 between the moving handle 23 and the operating ring 27. After the push handle 26 pushes the moving handle 23, the negative pressure spring 25 can cause the push handle 26 to spring back to its original position, thereby achieving a quantitative flushing.

[0039] The water collection tank is equipped with a hanging buckle 31 for suspending the tank. A button battery slot 18 is located on the outside of the tank for holding button batteries 19. Graduation lines 21 on the outer wall of the tank indicate the liquid content. A heating element 20 covers the outside of the tank, maintaining the liquid temperature at 37°C or other desired temperatures. A water inlet 22 and a negative pressure vent 23 are located on the surface of the tank to maintain a negative pressure state inside.

[0040] The fixed clamping ring 11 is connected to the fixed ring length adjusting buckle 15 via the fixed ring size adjustment line 14. The fixed clamping ring 11 and the fixed ring length adjusting buckle 15 are adjusted together via the fixed ring length adjusting line 29. One end of the fixed ring length adjusting line 29 is connected to the fixed ring length adjusting buckle 15 and extends to the top operating ring 27 of the push handle 26. The other end is connected to the fixed ring length adjusting wheel 30. Rotating the fixed ring length adjusting wheel 30 can change the length of the fixed ring length adjusting line 29, so that the fixed clamping ring 11 can move along the punch handle 12 with the fixed ring length adjusting buckle 15, thereby adjusting the distance between the fixed clamping ring 11 and the punch needle head.

[0041] During the embryo flushing operation, the embryo flushing needle 10 is inserted into the fallopian tube port, and the fallopian tube fixing clamp 11 can be wrapped around the outer wall of the fallopian tube port. By adjusting the size adjustment line 14 and the length adjustment line 29 of the fixing clamp, the size, length and position of the fixing clamp 11 in the fallopian tube can be adjusted, so that the end of the embryo flushing handle 12 and the fallopian tube opening are tightly fitted.

[0042] A semi-mechanized oviduct flushing device for in vivo fertilization in rodents, with a flushing needle 10, enables semi-mechanization of the oviduct in vivo fertilization process in rodents. It boasts advantages such as large water storage, quantitative flushing, and self-regulating temperature heating. This optimizes traditional operations, reduces workload, and simultaneously increases the number of embryos collected and the experimental output.

[0043] The procedure for using a semi-mechanized oviduct flushing device for in vivo fertilization in rodents includes the following steps:

[0044] 1. Install the movable fallopian tube placement dish 3 in the embryo flushing dish 1.

[0045] 2. Retrieve the collected fallopian tubes and place them in the center of the movable fallopian tube placement dish 3. Adjust the field of view under the microscope.

[0046] 3. Assemble the blank-punching needle 10, check whether the constant temperature heating device is operating normally, and fill the water collection tank with water.

[0047] 4. Locate the opening of the fallopian tube under a microscope, insert the embryo flushing needle 10, and adjust the size and length of the fixing ring 11 by adjusting the fixing ring size adjustment line 14 and the fixing ring length adjustment line 29 to fix the fallopian tube opening.

[0048] 5. By pushing the buckle 28 and the long handle 26, water in the water collection tank is flushed into the fallopian tube, thereby realizing the embryo flushing operation.

[0049] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A semi-mechanized oviduct flushing device for in vivo fertilization in rodents, comprising a flushing dish and a flushing apparatus, wherein the flushing dish is cylindrical and covered with a flushing cap; characterized in that, A movable fallopian tube placement dish is placed at the bottom of the embryo flushing dish. The fallopian tube is placed at the bottom of the movable fallopian tube placement dish. The movable fallopian tube placement dish includes an upper edge and a lower edge. The upper edge and the lower edge are connected by multiple connecting skeletons. A hollow groove is formed between the upper edge, the lower edge and the multiple connecting skeletons. Multiple support platforms are provided around the bottom of the movable fallopian tube placement dish, which can suspend and support the movable fallopian tube placement dish in the embryo flushing dish. The embryo flushing device includes an embryo flushing needle, a water collection tank, and a push handle. The embryo flushing needle and the water collection tank are connected through the embryo flushing needle interface. The embryo flushing needle is covered by an embryo flushing handle, one end of which is connected to the outer surface of the water collection tank. The needle tip of the embryo flushing needle extends beyond the other end of the embryo flushing handle. A soft plastic, adjustable-size fallopian tube fixing ring is fitted around one end of the embryo flushing handle near the embryo flushing needle. The inner ring of the fallopian tube fixing ring has fixing teeth for fixing the fixing ring to the outer wall of the fallopian tube port. The end of the water collection tank away from the embryo flushing needle is filled with a movable handle. The side of the movable handle away from the water collection tank is connected to the push handle. The end of the push handle away from the water collection tank has a push buckle. An operating ring is connected to the push handle near the push buckle. A negative pressure spring is fitted around the push handle between the movable handle and the operating ring.

2. The semi-mechanized oviduct flushing device for in vivo fertilization in rodents as described in claim 1, characterized in that, The movable fallopian tube placement dish is cylindrical.

3. The semi-mechanized oviduct flushing device for in vivo fertilization in rodents as described in claim 1, characterized in that, A heating coil is connected to the outside of the blanking pan. The heating coil is sleeved around the lower surface of the outer periphery of the blanking pan. The heating coil includes a heating coil body, a wire and a plug. The heating coil is a detachable external heater.

4. The semi-mechanized oviduct flushing device for in vivo fertilization in rodents as described in claim 1, characterized in that, The tooth profile of the fixed clamping teeth is an obtuse triangle.

5. The semi-mechanized oviduct flushing device for in vivo fertilization in rodents as described in claim 1, characterized in that, The blanking tool also includes a fixed ring size adjustment line and a fixed ring length adjustment buckle. The fixed clamping ring is connected to the fixed ring length adjustment buckle through the fixed ring size adjustment line. The fixed ring length adjustment buckle is fitted onto the long handle of the blanking tool. The fixed ring length adjustment buckle is provided with fixed ring length adjustment teeth, which can extend or contract the fixed ring size adjustment line and wrap it around the fixed ring length adjustment teeth to adjust the length and width of the fixed ring size adjustment line, thereby adjusting the size of the fixed clamping ring.

6. The semi-mechanized oviduct flushing device for in vivo fertilization in rodents as described in claim 5, characterized in that, The blank punch also includes a fixed ring length adjustment tooth and a fixed ring length adjustment line. The operating ring is provided with a fixed ring length adjustment wheel. One end of the fixed ring length adjustment line is connected to the fixed ring length adjustment buckle and extends to the operating ring at the top of the push handle. The other end of the fixed ring length adjustment line is connected to the fixed ring length adjustment wheel. Rotating the fixed ring length adjustment wheel can change the length of the fixed ring length adjustment line, so that the fixed clamping ring can move along the blank punch handle with the fixed ring length adjustment buckle, thereby adjusting the distance between the fixed clamping ring and the blank punch needle head.

7. The semi-mechanized oviduct flushing device for in vivo fertilization in rodents as described in claim 1, characterized in that, The water collection tank is equipped with a button battery slot for holding button batteries. The outer wall of the water collection tank is equipped with scale lines to indicate the liquid content inside the water collection tank. The water collection tank is covered with a heating plate to maintain the liquid inside the water collection tank at the required temperature. The surface of the water collection tank is equipped with a water inlet and a negative pressure vent to maintain the negative pressure state inside the water collection tank.

Images