Sperm preferred dish and sperm capacitation liquid preloaded device

Abstract

Sperm preferred culture dish and sperm capacitation liquid preloading device relate to the technical field of sperm culture dish, and are used for providing a culture dish with sperm preferred function and a device convenient for preloading capacitation liquid. The sperm preferred culture dish comprises a culture dish main body and a limiting buckle, and the culture dish main body is internally provided with a capacitation liquid adding hole, a sperm adding hole and a sperm collecting hole. The sperm capacitation liquid preloading device comprises a preloader and a movable rod, the preloader is movably arranged above the culture dish, the edge of the preloader is fixed with a supporting column, and the bottom of the supporting column is provided with a moving wheel. The top of the preloader is provided with a liquid injection port; the movable rod movably penetrates through the preloader, the movable rod and the preloader are provided with a spring therebetween, and the upper end of the movable rod is fixed with a limiting plate. The sperm preferred culture dish and the sperm capacitation liquid preloading device can realize the selection of sperm and preloading of sperm capacitation liquid.

Description

Technical Field

[0001] This invention relates to the field of sperm culture dish technology, specifically to a sperm selection culture dish and a sperm capacitation fluid pre-loading device. Background Technology

[0002] Sperm culture requires the use of culture dishes for subsequent biological experiments. To improve sperm culture, sperm capacitation medium needs to be added to the culture dishes. Therefore, a pre-filled capacitation medium device needs to be added to the culture dishes. For example, a culture device for intracytoplasmic sperm injection (ICSI) published in CN217868910U includes a culture dish body, a fixing sleeve, and a supporting sleeve. This improves the stability of the culture dish during use, making it less likely to shift or fall off due to external collisions or vibrations. Furthermore, the culture dish is not easily detached after assembly, making it convenient to use. However, this ICSI culture device still has the following drawbacks in practical use: although the above-mentioned culture device achieves sperm culture, it does not have a sperm selection function. Sperm capacitation medium needs to be added to the sperm selection culture dish during use, making the entire operation relatively cumbersome. Summary of the Invention

[0003] The purpose of this invention is to provide a sperm selection culture dish and a sperm capacitation fluid pre-loading device to solve the problems mentioned in the background art, such as the lack of sperm selection function in the culture dish and the inability to pre-load sperm capacitation fluid.

[0004] The technical solution adopted by the present invention to solve its technical problem is: a sperm selection culture dish, including a culture dish body, the bottom of the culture dish body has a sperm addition hole and a capacitation fluid addition hole, the capacitation fluid addition holes are evenly distributed on the circumference with the sperm addition hole as the center, the bottom of the sperm addition hole is lower than the bottom of the capacitation fluid addition hole, and the bottom of the sperm addition hole is connected to the bottom of the capacitation fluid addition hole through an inclined flow channel, the upper middle part of the flow channel is recessed downward to form a sperm collection hole.

[0005] Furthermore, a limit buckle is fixed to the side wall of the culture dish body.

[0006] The present invention also provides a sperm capacitation fluid preloading device comprising a sperm selection culture dish, including a preloading device and a movable rod. The preloading device is movably disposed above the main body of the culture dish. A support column is fixed at the edge of the preloading device, and a movable wheel is movably installed at the bottom of the support column. The support column is snapped into connection with a limiting buckle and can rotate relative to the limiting buckle. A liquid injection port is provided at the top of the preloading device, and liquid storage chambers are evenly distributed inside the preloading device. A liquid outlet hole is reserved at the bottom of the liquid storage chamber. The movable rod is movably connected to the preloading device and passes through the preloading device from top to bottom. A limiting plate is fixed at the upper end of the movable rod. A fixing groove is provided in the upper surface of the preloading device, and a spring is loosely fitted on the outside of the movable rod between the limiting plate and the bottom of the fixing groove.

[0007] Furthermore, the bottom of the moving wheel is flush with the bottom surface of the culture dish body.

[0008] Furthermore, the connection between the energy-generating liquid addition hole and the flow channel has a flow-limiting mesh.

[0009] Furthermore, the liquid storage chamber and the liquid outlet are both distributed at equal angles on the pre-loading device, and the energy harvesting liquid in the liquid storage chamber falls into the energy harvesting liquid addition hole on the main body of the culture dish through the liquid outlet.

[0010] Furthermore, the movable rod and the pre-installer are slidably connected, the movable rod and the limiting plate are perpendicular to each other, the length of the limiting plate is less than the length of the fixed groove, and the width of the limiting plate is less than or equal to the width of the fixed groove.

[0011] Furthermore, a connecting plate is rotatably mounted on the bottom of the movable rod, and a sealing block is fixed at the edge of the connecting plate.

[0012] Furthermore, the sealing block has a frustum-shaped structure, and the sealing block and the liquid outlet are arranged in a one-to-one correspondence, with the shape of the liquid outlet adapted to the shape of the sealing block.

[0013] Compared with the prior art, the beneficial effects of the present invention are: (1) The pre-loading device and the sperm selection culture dish are detachably connected, which facilitates the alignment and adjustment of the pre-loading device and the sperm selection culture dish, ensuring that the liquid storage chamber and the capacitation liquid addition hole are aligned vertically, thus providing a guarantee for the pre-loading of the capacitation liquid.

[0014] (2) When the movable rod is rotated 90° and pushed downward, the limiting plate can be moved in the fixed groove, which in turn drives the connecting plate to move downward, releasing the sealing block from the liquid outlet. At this time, the energy-generating liquid in the storage chamber can be discharged into the energy-generating liquid addition hole to realize the quantitative pre-filling of the energy-generating liquid. Attached Figure Description

[0015] Figure 1 This is a front view structural diagram of the present invention; Figure 2 This is a front view schematic diagram of the limiting buckle structure of the present invention; Figure 3 This is a schematic diagram of the front section structure of the present invention; Figure 4 For the present invention Figure 3 Enlarged structural diagram at point A in the middle; Figure 5 This is a top-section schematic diagram of the pre-assembler of the present invention; Figure 6 This is a top view of the connecting plate structure of the present invention; Figure 7 This is a schematic diagram of the pre-assembler structure after rotation. Figure 8 Top view of a culture dish for sperm selection; Figure 9 for Figure 8 BB section view in the middle; In the diagram: 1. Petri dish body; 2. Limiting buckle; 21. Notch; 3. Pre-loading device; 4. Support column; 5. Casters; 6. Injection port; 7. Storage chamber; 8. Outlet hole; 9. Movable rod; 91. Handle; 10. Limiting plate; 11. Fixing groove; 12. Connecting plate; 13. Sealing block; 14. Capacitating fluid addition hole; 15. Sperm addition hole; 16. Flow channel; 17. Sperm collection hole; 18. Spring; 19. Flow limiting net. Detailed Implementation

[0016] like Figure 1 , Figure 3 , Figure 8 and Figure 9 As shown, the sperm selection culture dish includes a culture dish body 1 and a limiting buckle 2, which is fixed to the side of the culture dish body 1. The culture dish body 1 is a cylindrical hollow structure with an open top. The limiting buckle 2 is also cylindrical, with a C-shaped notch 21 that penetrates the upper and lower surfaces of the limiting buckle 2 and is located on the side wall of the limiting buckle 2. Figure 8As shown, the bottom of the culture dish body 1 has capacitation fluid addition holes 14 and sperm addition holes 15. Three capacitation fluid addition holes 14 are evenly arranged circumferentially. The sperm addition holes 15 are located at the center of the culture dish body 1, and the capacitation fluid addition holes 14 are located at the edge of the culture dish body 1, specifically on a circle centered at the sperm addition holes 15. The capacitation fluid addition holes 14 and sperm addition holes 15 are connected by a flow channel 16. The flow channel 16 is inclined, with its upper end connected to the bottom of the capacitation fluid addition hole 14 and its lower end connected to the bottom of the sperm addition hole 15. Thus, after adding sperm capacitation fluid to the capacitation fluid addition hole 14, the sperm capacitation fluid can flow through the flow channel 16 to the sperm addition hole 15. A flow-limiting mesh 19 is provided between the upper end of the flow channel 16 and the capacitation fluid addition hole 14. The flow-limiting mesh 19 has precise micropores calculated based on the composition of the capacitation fluid and fluid tension dynamics, which can control the slow flow of the capacitation fluid towards the sperm addition hole 15. The inclination angle of the flow channel 16 is 30-45 degrees, and the middle of the flow channel 16 is recessed downward to form a sperm collection hole 17. The depth of the sperm collection hole 17 is 10-100 micrometers, which can ensure the smooth flow of the capacitation fluid and maximize the collection of high-quality sperm from upstream. In use, semen is added to the sperm addition hole 15, and then capacitation fluid is added to the capacitation fluid addition hole 14. The capacitation fluid flows slowly along the flow channel 16 to the sperm addition hole 15. At this time, the sperm in the sperm addition hole 15, under the stimulation of the capacitation fluid, the inclined design of the flow channel 16, and the forward motility and tropism of the sperm itself, flow counterclockwise along the flow channel 16 from low to high. High-quality sperm will gather in the sperm collection hole 17. Multiple capacitation fluid addition holes 14 are evenly arranged on the same circumference, such as three, which can increase the selectivity of sperm swimming upward and facilitate sperm selection.

[0017] Pre-filled devices for sperm capacitation fluid containing sperm selection culture dishes, such as Figure 1 As shown, the device includes a pre-loading device 3 and a movable rod 9. The pre-loading device 3 is movably positioned above the main body 1 of the culture dish, with its bottom contacting the top of the main body 1. A support column 4 is fixed to the edge of the pre-loading device 3, and a caster wheel 5 is movably mounted on the bottom of the support column 4. The support column 4 extends into the notch 21 of the limiting buckle 2 to achieve a snap-fit ​​connection with the limiting buckle 2, and the support column 4 can rotate relative to the limiting buckle 2. The top of the pre-loading device 3 is provided with several liquid injection ports 6, such as... Figure 3 As shown, the pre-loading device 3 has a liquid storage chamber 7 inside, such as... Figure 4 As shown, the bottom of the storage chamber 7 has a pre-reserved outlet hole 8. The outlet hole 8 is a conical hole, and its diameter increases from top to bottom. The number of injection ports 6, storage chambers 7, and outlet holes 8 are the same and correspond one-to-one. The injection ports 6, storage chambers 7, and outlet holes 8 are arranged sequentially from top to bottom, and the injection ports 6 are connected to the outlet holes 8 through the storage chambers 7. Insert the support column 4 into the notch 21 on the limiting buckle 2 of the sperm selection culture dish, and align the pre-loading device 3 with the sperm selection culture dish vertically, so that the liquid storage chamber 7 is aligned vertically with the capacitating liquid addition hole 14.

[0018] like Figure 1 , Figure 3 As shown, the movable rod 9 extends through the top of the pre-assembler 3. The top of the movable rod 9 has a handle 91. A spring 18 is wound around the middle of the movable rod 9. A limit plate 10 is fixed to the upper end of the movable rod 9. A fixing groove 11 is formed in the pre-assembler 3 below the limit plate 10. The lower end of the spring 18 contacts and is fixed to the bottom of the fixing groove 11, and the upper end of the spring 18 contacts the bottom of the limit plate 10. The movable rod 9 and the pre-assembler 3 are slidably connected, and the movable rod 9 and the limit plate 10 are perpendicular to each other. Figure 1 As shown, the length of the limiting plate 10 is less than the length of the fixing groove 11, and the width of the limiting plate 10 is less than or equal to the width of the fixing groove 11. Figure 6 As shown, the bottom of the movable rod 9 is connected to a connecting plate 12 via a bearing, and a sealing block 13 is fixed at the edge of the connecting plate 12. The sealing block 13 has a frustum-shaped structure, and the number of sealing blocks 13 is the same as the number of liquid outlet holes 8, and the sealing blocks 13 and liquid outlet holes 8 are set in a one-to-one correspondence.

[0019] like Figure 1 , Figure 2 and Figure 7 As shown, the bottom of the movable wheel 5 is flush with the bottom surface of the culture dish body 1. The support column 4 is used to connect with the limiting buckle 2. The support column 4 passes through the notch 21 on the limiting buckle 2, so the support column 4 can rotate relative to the limiting buckle 2. When the pre-loading device 3 is rotated, the support column 4 rotates relative to the limiting buckle 2, and the pre-loading device 3 rotates on the culture dish body 1 to ensure that the pre-loading device 3 is aligned vertically with the sperm selection culture dish, thus facilitating the accurate falling of the sperm capacitation fluid in the storage chamber 7 into the capacitation fluid addition hole 14. During the rotation of the pre-loading device 3, the movable wheel 5 can be driven to roll on the table, thereby improving the smoothness of rotation and ensuring the stability of the pre-loading device 3 after rotation, preventing it from tipping over. In use, the sperm selection culture dish is placed under the pre-loading device 3. Then, the support column 4 is inserted into the notch 21 of the limiting buckle 2. On the one hand, it is convenient to adjust the pre-loader 3 so that the pre-loader 3 and the sperm selection culture dish are aligned vertically. On the other hand, when rotating and pressing the handle 91 at the top of the movable rod 9, the cooperation between the support column 4 and the limiting buckle 2 can prevent the pre-loader 3 and the sperm selection culture dish from being misaligned at a large angle.

[0020] like Figure 5 As shown, the liquid storage chamber 7 and the liquid outlet 8 are both distributed at equal angles on the pre-loading device 3, such as... Figure 3As shown, the liquid storage chamber 7 is interconnected with the interior of the culture dish body 1 through the liquid outlet 8, and the number of liquid storage chambers 7 is the same as the number of capacitating liquid addition holes 14. After rotating the pre-loading device 3 so that the pre-loading device 3 is vertically aligned with the culture dish body 1, the liquid outlet 8 is vertically aligned with the corresponding capacitating liquid addition hole 14, that is, there is a liquid outlet 8 above each capacitating liquid addition hole 14.

[0021] When adding capacitating fluid, first add sperm capacitating fluid into the storage chamber 7 through the injection port 6. Then rotate the movable rod 9 by 90°. As the movable rod 9 rotates, it drives the limiting plate 10 to rotate synchronously until the limiting plate 10 is directly above the fixed groove 11, and the limiting plate 10 and the positioning groove 11 are aligned vertically. At this time, the connecting plate 12 remains stationary. Then press down on the handle 91 of the movable rod 9. At this time, the movable rod 9 slides downward relative to the pre-loading device 3, and drives the limiting plate 10 to slide in the fixed groove 11. At the same time, it drives the connecting plate 12 to descend synchronously, so that the upper end of the sealing block 13 moves out of the liquid outlet 8, thereby releasing the sealing block 13 from the liquid outlet 8. At this time, the capacitating fluid in the storage chamber 7 is discharged into the capacitating fluid addition hole 14 through the liquid outlet 8. Since the volume of the storage chamber 7 is fixed, the capacitating fluid can be added quantitatively. When the pressure on the top handle 91 of the movable rod 9 is released, the spring 18 lifts the limiting plate 10, which in turn drives the connecting plate 12 to rise and causes the sealing block 13 to re-block the liquid outlet 8. The movable rod 9 is rotated in the opposite direction so that the limiting plate 10 is misaligned with the positioning groove 11, thereby limiting the movable rod 9 through the limiting plate 10 to prevent accidental contact with the movable rod 9.

[0022] The method of using this invention is described below: (1) Inject sperm capacitation fluid into the storage chamber 7 inside the pre-loading device 3 through the injection port 6; (2) Insert the support column 4 on the pre-loading device 3 into the notch 21 of the limiting buckle 2 of the sperm selection culture dish; (3) Rotate the pre-loading device 3 around the support column 4 so that the pre-loading device 3 is aligned vertically with the main body 1 of the culture dish, and at this time the liquid outlet 8 is aligned vertically with the energy collection liquid addition hole 14. (4) Rotate the handle 91 so that the limiting plate 10 is aligned with the positioning groove 11; then press the handle 91 down so that the connecting plate 12 moves down, thereby releasing the sealing block 13 from the liquid outlet hole 8. At this time, the energy harvesting liquid in the storage chamber 7 flows into the energy harvesting liquid addition hole 14 of the culture dish body 1 through the liquid outlet hole 8. (5) After the addition is completed, release the pressure on the handle 91. Under the action of the spring 18, the sealing block 13 resumes the sealing of the liquid outlet 8. Then rotate the handle 91 so that the limiting plate 10 and the positioning groove 11 are misaligned. (6) Separate the sperm selection culture dish pre-filled with capacitation solution from the support column 4 to achieve the pre-filling of sperm capacitation solution.

[0023] The sperm capacitation solution of this invention is divided into solution A and solution B. Solution A is the basic culture medium for in vitro sperm manipulation, and its components include: sodium chloride 80-100 mM, potassium chloride 3-8 mM, magnesium sulfate 0.5-1.5 mM, sodium dihydrogen phosphate 0.05-1.5 mM, potassium dihydrogen phosphate 0.05-1.5 mM, calcium chloride 0.5-2.5 mM, sodium bicarbonate 10-30 mM, HEPES sodium salt 10-30 mM, glucose 0.05-5.0 mM, sodium lactate 5.0-15 mM, sodium pyruvate 0.1-1.0 mM, taurine 0.1-10 mM, and human serum albumin 2-10 mg / ml. Fluid B is a capacitation factor, and its components include one or more of the following: angiotensin II 10-100 mM, calcitonin 0.01-15 nM, pentoxifylline 0.7-7 mM, and antioxidants. The antioxidants include: glutathione 1-10 mM, vitamin E 2-20 nM, acetyl-L-carnitine 1-15 μM, acetylcysteine ​​1-20 μM, α-lipoic acid 1-10 μM, and platelet-activating factor 15-90 nM.

[0024] The advantages of the present invention are: (1) The pre-loading device and the sperm selection culture dish are detachably connected, which facilitates the alignment adjustment of the pre-loading device and the sperm selection culture dish, ensuring the vertical alignment of the storage chamber and the capacitation liquid addition hole, thus providing a guarantee for the pre-loading of the capacitation liquid; in addition, the assembly method of the pre-loading device and the sperm selection culture dish can also avoid large-angle misalignment between the pre-loading device and the sperm selection culture dish during the operation of pre-loading; (2) When the movable rod is rotated 90° and pushed downward, the limiting plate can be moved in the fixed groove, thereby moving the connecting plate downward, releasing the sealing block from the liquid outlet hole, and at this time the capacitation liquid in the storage chamber can be discharged into the capacitation liquid addition hole to realize the quantitative pre-loading of the capacitation liquid.

Claims

1. A sperm capacitation fluid pre-loading device, characterized in that, The system includes a sperm selection culture dish, a pre-loading device (3), and a movable rod (9). The sperm selection culture dish includes a culture dish body (1). The bottom of the culture dish body (1) has a sperm addition hole (15) and a capacitation fluid addition hole (14). The capacitation fluid addition holes (14) are evenly distributed on a circumference with the sperm addition hole (15) as the center. The bottom of the sperm addition hole (15) is lower than the bottom of the capacitation fluid addition hole (14). The bottom of the sperm addition hole (15) is connected to the bottom of the capacitation fluid addition hole (14) through an inclined flow channel (16). The upper middle part of the flow channel (16) is recessed downward to form a sperm collection hole (17). The side wall of the culture dish body (1) is fixed with a limiting buckle (2). The connection between the capacitation fluid addition hole (14) and the flow channel (16) has a flow-limiting mesh (19). The pre-loading device ( 3) The movable device is located above the main body of the petri dish (1). A support column (4) is fixed at the edge of the pre-loading device (3). A movable wheel (5) is movably installed at the bottom of the support column (4). The support column (4) is snapped into the limit buckle (2) and the support column (4) can rotate relative to the limit buckle (2). The top of the pre-loading device (3) is provided with a liquid injection port (6). The interior of the pre-loading device (3) is evenly distributed with liquid storage chambers (7). The bottom of the liquid storage chambers (7) is reserved with a liquid outlet hole (8). The movable rod (9) is movably connected to the pre-loading device (3) and passes through the pre-loading device (3) from top to bottom. A limit plate (10) is fixed at the upper end of the movable rod (9). The upper surface of the pre-loading device (3) has a fixing groove (11). A spring (18) is loosely fitted on the outside of the movable rod (9) between the limit plate (10) and the bottom of the fixing groove (11). A connecting plate (12) is rotatably mounted on the bottom of the movable rod (9), and a sealing block (13) is fixed at the edge of the connecting plate (12); the sealing block (13) is a frustum-shaped structure, and the sealing block (13) and the liquid outlet (8) are arranged in a one-to-one correspondence, and the shape of the liquid outlet (8) is adapted to the shape of the sealing block (13).

2. The sperm capacitation fluid pre-loading device according to claim 1, characterized in that, The bottom of the moving wheel (5) is flush with the bottom surface of the petri dish body (1).

3. The sperm capacitation fluid pre-loading device according to claim 1, characterized in that, The liquid storage chamber (7) and the liquid outlet (8) are both distributed at equal angles on the pre-loading device (3), and the energy-collecting liquid in the liquid storage chamber (7) falls into the energy-collecting liquid addition hole (14) on the main body of the culture dish (1) through the liquid outlet (8).

4. The sperm capacitation fluid pre-loading device according to claim 1, characterized in that, The movable rod (9) and the pre-installer (3) are slidably connected. The movable rod (9) and the limiting plate (10) are perpendicular to each other. The length of the limiting plate (10) is less than the length of the fixed groove (11). The width of the limiting plate (10) is less than or equal to the width of the fixed groove (11).

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