Flusher for collecting ovum or early embryo in murine fallopian tube
By designing a flushing device for collecting oocytes or early embryos from the fallopian tubes of mice and using culture medium flushing, the problems of low efficiency and sample damage in existing technologies have been solved, achieving efficient and non-damaging oocyte and embryo collection, which meets the needs of biomedical research.
Patent Information
- Application Number
- CN202422767795.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-13
- Publication Date
- 2025-12-02
- Estimated Expiration
- 2034-11-13
AI Technical Summary
Existing techniques are difficult to efficiently obtain oocytes and early embryos from the fallopian tubes of mice, and the operation is inefficient, easily damaging the samples and affecting subsequent research.
An irrigator comprising components such as an irrigating needle, an irrigating hose, and a pipette nozzle is used to achieve efficient collection of oocytes and embryos from the fallopian tubes through micromanipulation and irrigation with culture medium. An irrigator designed to solve this problem includes components such as a protective cap, an irrigating needle, a pipette nozzle, and an irrigation culture medium sample loading port, which utilizes the pressure of the culture medium to obtain oocytes and embryos.
This method enables efficient and non-destructive acquisition of oocytes and embryos from the oviducts of mice, improving recovery rate and sample integrity, and meeting the needs of subsequent research.
Smart Images
Figure CN223614973U_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of laboratory equipment for rodents, and in particular to a flushing device for collecting oocytes or early embryos from the fallopian tubes of rodents. Background Technology
[0002] Mouse eggs and early embryos have a wide range of applications in biomedical research, including: genetics and developmental studies, transgenic animal production, research on the function of developmental genes, drug screening, and toxicity testing.
[0003] Rodents are model mammals for understanding, and can be used to construct models of reproductive, developmental, and disease mechanisms. Mature oocytes and early embryos in rodents are located in the oviduct. Isolating mature oocytes and embryos from the oviduct is an indispensable step in these studies.
[0004] Current techniques can only retrieve oocytes from specific locations in the fallopian tubes, or are only applicable to specific strains of mice. Furthermore, the process is inefficient, resulting in poor integrity of the obtained oocytes and early embryos, which are easily damaged during the procedure, hindering subsequent research.
[0005] The information disclosed in the background section is intended only to enhance the understanding of the overall background of this utility model and should not be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art. Utility Model Content
[0006] This application addresses the aforementioned technical problem by providing a flushing device for collecting oocytes or early embryos from the fallopian tubes of rodents, which can be used to recover all embryos located at different positions in the fallopian tubes and at different developmental stages.
[0007] This application provides a flushing device for collecting oocytes or early embryos from the oviducts of mice, comprising: an operating handle, a flushing hose, a flushing culture medium application port, a pipette tip, and a flushing needle;
[0008] The flushing needle is detachably mounted on one end of the operating handle; the flushing hose is inserted into the operating handle and connected to the flushing needle.
[0009] The other end of the rinsing hose is equipped with a rinsing culture medium application port and a pipette tip at intervals.
[0010] Preferably, it includes: a piston; the piston is disposed on one end of the flushing hose; the piston is fixedly disposed inside the operating handle.
[0011] Preferably, the flushing needle includes: a connecting section and an extension end; the extension end forms an angle β with the connecting section; the angle β = 100° to 120°.
[0012] Preferably, it includes: a needle plug; a fixed end of a flushing needle is mounted on the needle plug; an interface is provided on one end of the operating handle; the needle plug is disassembled and inserted into the interface.
[0013] Preferably, it includes: a protective cap; the protective cap is detached and fitted onto the flushing needle.
[0014] Preferably, it includes: a filter head; the filter head is housed in one end of the rinsing hose and positioned near the suction nozzle.
[0015] Preferably, the flushing hose is a PVC hose.
[0016] Preferably, the diameter of the rinsing needle is 0.4 mm, 0.45 mm or 0.5 mm.
[0017] Preferably, the extended end of the rinsing needle is flat.
[0018] Preferably, the length of the rinsing needle is less than or equal to 30 mm.
[0019] The beneficial effects that this application can produce include:
[0020] 1) The flushing device provided in this application for collecting oocytes or early embryos in the oviducts of mice can quickly retrieve oocytes (embryos) from the oviducts of mice when using this device. The operation is simple, does not damage the sample, and has a high recovery rate.
[0021] 2) The flushing device provided in this application for collecting oocytes or early embryos in the fallopian tubes of mice can avoid damage to the mice and prevent death from non-experimental factors when using this device to collect oocytes and early embryos. The collected samples have high integrity and meet the requirements of subsequent research experiments. Attached Figure Description
[0022] Figure 1 A schematic diagram of the main structure of a flushing device for collecting oocytes or early embryos from the fallopian tubes of mice, provided in at least one embodiment of this application;
[0023] Legend:
[0024] 1. Protective cap; 2. Flushing needle; 3. Needle plug; 5. Piston; 4. Connector; 6. Operating handle; 7. PVC flushing hose; 8. Flushing culture medium sample dispensing port; 9. Filter head; 10. Pipette nozzle. Detailed Implementation
[0025] To make the objectives, technical solutions, and advantages of the embodiments of this utility model clearer, the technical solutions of the embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this utility model, not all embodiments. The components of the embodiments of this utility model described and shown in the accompanying drawings can typically be arranged and designed in various different configurations.
[0026] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.
[0027] Technical means not detailed in this application and not used to solve the technical problems of this application are all set according to common general knowledge in the field, and multiple common general knowledge setting methods can be implemented.
[0028] See Figure 1 The flushing device provided in this application for collecting oocytes or early embryos from the fallopian tubes of mice includes: a protective cap 1, a flushing needle 2, a needle plug 2, a connector 2, a piston 5, an operating handle 6, a PVC flushing hose 7, a flushing culture medium application port 8, a filter head 9, and a pipette tip 10. The flushing culture medium is added to the PVC flushing hose 7 through the flushing culture medium application port 8. Under a stereomicroscope, the extension end of the flushing needle 2 is inserted into the fallopian tube of the mouse through micromanipulation.
[0029] The operator blows air into the PVC flushing tube 7 through the pipette nozzle 10, driving the culture medium inside the PVC flushing tube 7 to flush out the oocytes (embryos) located in the fallopian tubes, thus retrieving the oocytes (embryos). A sample inlet 8 for adding flushing culture medium is provided on one side of the air inlet of the PVC flushing tube 7, allowing for easy addition of flushing culture medium as needed during the operation. The pressure driving the culture medium is created by the operator blowing air into the PVC flushing tube 7 through the pipette nozzle 10, freeing the operator's hands and facilitating micromanipulation.
[0030] Washing with culture medium can avoid contamination by other substances and improve the reliability of the results.
[0031] The device has a flushing needle 2 with a flat end. The flushing needle 2 is at an angle β to the connecting section, with the angle β being 100° to 120°. This allows the flushing needle 2 to bend in a way that fits the mouse's body structure, facilitating the insertion of the needle into the mouse to retrieve eggs and perform early embryo manipulation.
[0032] The PVC flushing hose is fixed to the connector of the operating handle by a needle plug. It is sealed and fixed inside the operating handle by a piston and has a sample inlet for flushing the culture medium. A pipette tip with a filter is connected to the distal end of the PVC flushing hose. Oocytes (embryos) are retrieved by the operator blowing air into the PVC flushing hose to pressurize the flushing medium, flushing out mature ovums (embryos) located in the oviduct. This instrument can efficiently recover mature ovums (embryos) from the oviducts of rodents, is simple to operate, causes no damage to the sample, and has a significant recovery effect.
[0033] In one specific embodiment, the flushing needle 2 is inserted into the connector 4 of the operating handle 6 via a needle plug 3. The specification of the flushing needle 2 can be selected according to the inner diameter of the oviduct lumen of different rodents, and it is convenient to replace. Specifically, the diameter of the flushing needle 2 is 0.4 mm, 0.45 mm, or 0.5 mm.
[0034] In one specific embodiment, a piston 5 is provided on the outer wall of one end of the PVC rinsing hose 7. The piston 5 and part of the PVC rinsing hose 7 are accommodated within the operating handle 6. The piston 5 is fixedly disposed within the operating handle 6 to seal the outer wall of the PVC rinsing hose 7 and the inner wall of the operating handle 6, and to fix the position of the PVC rinsing hose 7 in the operating handle 6. It also connects the PVC rinsing hose 7 with the connector 4 to achieve directional flow of the culture medium. The piston 5 used is made of silicone.
[0035] In one specific embodiment, a filter head 9 is housed in the air inlet of the PVC rinsing hose 7; a pipette nozzle 10 is provided on the air inlet of the PVC rinsing hose 7 to facilitate air blowing by the operator and to prevent impurities in the gas from contaminating the culture medium. The filter head 9 is made of high molecular weight polypropylene material and can be inserted into the PVC rinsing hose 7.
[0036] In one specific embodiment, it includes: a protective cap 1; the protective cap 1 is detachably fitted onto the flushing needle 2 of the operating handle 6. This facilitates the cleaning of the flushing needle 2.
[0037] In one specific embodiment, the length of the irrigation needle 2 is less than or equal to 30 mm to avoid puncturing the organ while ensuring that the irrigation fluid can reach the irrigation position.
[0038] This new instrument can efficiently recover mature eggs and embryos from the fallopian tubes. It is simple to operate and does not damage the sample.
[0039] Application Example: In August 2024, an embryo culture environment test was conducted in the Human Embryology Laboratory of the Department of Reproductive Medicine at the Second Affiliated Hospital of Kunming Medical University. The test used mouse blastocyst formation rate as the evaluation index. Mouse blastocysts needed to be collected.
[0040] Using the flushing device with the above structure, the fallopian tubes obtained from the dissection of laboratory mice were manipulated to obtain mouse blastocysts.
[0041] Mice were superstimulated to ovulate by intraperitoneal injection of pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (HCG). Mice were euthanized by cervical dislocation 36 hours post-mating, and the oviducts were dissected. Using a flushing device with the aforementioned structure, under a stereomicroscope, the curved end of the flushing needle 2 was inserted into the mouse oviduct. The operator blew air through the air inlet of the PVC flushing tubing 7, driving the culture medium (GMOPS-plus) within the tubing to flush out the 2-cell embryos located in the oviduct. After washing in GMOPS-plus, the obtained embryos were transferred to TL medium for blastocyst culture.
[0042] This experiment used two strains of mice (Kunming mice and C57BL / 6) and was repeated three times.
[0043] The experiment was repeated three times. The average blastocyst formation rate was 96%.
[0044] The table below lists the blastocyst retrieval results from the three experiments:
[0045] surface
[0046] Number of 2-cell embryos Number of blastocysts blastocyst rate First experiment 49 47 0.96 Second experiment 37 35 0.95 Third experiment 26 26 1 total 112 108 0.96
[0047] This flushing device is convenient and quick for obtaining mouse 2-cell embryos, and the experimental procedure does not cause any damage to the developmental potential of the mouse embryos.
[0048] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A flushing device for collecting oocytes or early embryos from the fallopian tubes of rodents, characterized in that, include: Operating handle (6), flushing hose, flushing culture medium sample dispensing port (8), pipette tip (10), flushing needle (2); The flushing needle (2) is detached and installed on one end of the operating handle (6); the flushing hose is inserted into the operating handle (6) and connected to the flushing needle (2); The other end of the rinsing hose is provided with a rinsing culture medium sample inlet (8) and a pipette tip (10) at intervals.
2. The flushing device for collecting oocytes or early embryos from the fallopian tubes of mice according to claim 1, characterized in that, include: Piston (5); The piston (5) is located at one end of the flushing hose; The piston (5) is fixedly installed inside the operating handle (6).
3. The flushing device for collecting oocytes or early embryos from the fallopian tubes of mice according to claim 1, characterized in that, The flushing needle (2) includes: a connecting section and an extension end; the extension end forms an angle β with the connecting section; the angle β = 100° to 120°.
4. The flushing device for collecting oocytes or early embryos from the fallopian tubes of mice according to claim 3, characterized in that, include: Needle plug (3); the fixed end of the flushing needle (2) is installed on the needle plug (3); an interface (4) is provided on one end of the operating handle (6); the needle plug (3) is disassembled and inserted into the interface (4).
5. The flushing device for collecting oocytes or early embryos from the fallopian tubes of mice according to claim 1, characterized in that, include: Protective cap (1); the protective cap (1) is removed and fitted onto the flushing needle (2).
6. The flushing device for collecting oocytes or early embryos from the fallopian tubes of mice according to claim 1, characterized in that, include: Filter head (9); The filter head (9) is housed in one end of the rinsing hose and is positioned near the suction nozzle (10).
7. The flushing device for collecting oocytes or early embryos from the fallopian tubes of mice according to claim 1, characterized in that, The flushing hose is a PVC hose.
8. The flushing device for collecting oocytes or early embryos from the fallopian tubes of mice according to claim 1, characterized in that, The rinsing needle (2) has a diameter of 0.4 mm, 0.45 mm or 0.5 mm.
9. The flushing device for collecting oocytes or early embryos from the fallopian tubes of mice according to claim 1, characterized in that, The extended end of the rinsing needle (2) is flat.
10. The flushing device for collecting oocytes or early embryos from the fallopian tubes of mice according to claim 1, characterized in that, The length of the rinsing needle (2) is less than or equal to 30 mm.