A vessel for conveniently observing the morphology of oocytes and its usage method

Abstract

The present invention discloses a vessel for conveniently observing the morphology of oocytes and a using method thereof. The vessel includes a circular concave hole, a long strip-shaped groove, a non-connected groove, and a circular groove. The non-connected groove and the circular groove are arranged on the outer side of the circular concave hole. The long strip-shaped groove is arranged between the circular concave hole and the circular groove. The circular concave hole and the circular groove are communicated through the long strip-shaped groove. The shape of the non-connected groove is circular, and the center of the non-connected groove and the center of the circular concave hole are on a straight line. The present invention mainly uses the surface tension of the interface between the culture solution and the mineral oil to slightly squeeze the OCC, pushing the granulosa cells to the periphery of the oocyte, and a relatively complete oocyte morphology can be observed. In particular, the GV-stage oocytes can be clearly distinguished. For the first time, a simple method for observing the maturity of oocytes without removing granulosa cells is proposed.

Description

Technical Field

[0001] The present invention relates to the technical field of oocyte observation vessels, and particularly to a vessel for conveniently observing the morphology of oocytes and a using method thereof. Background Art

[0002] In assisted reproductive technology, in vitro fertilization of oocytes is often carried out, and the maturity of oocytes will directly affect the fertilization outcome. The maturity of oocytes is divided into three degrees: MⅡ stage, the nucleus disappears and the first polar body is released; MⅠ stage, the nucleus disappears and the first polar body has not been discharged; GV stage, nuclear structure can be seen in the cytoplasm. Among them, the MⅡ stage is mature oocytes with the highest fertilization rate, while GV stage oocytes can hardly be fertilized. The complex collected during oocyte retrieval surgery is called oocyte corona-cumulus complex (OCC), which consists of oocytes with a diameter of 100-150 μm, the corona radiata surrounding the oocytes and cumulus cells. The corona radiata is composed of inner granular cells, and the cumulus cells are outer granular cells. However, OCC will affect the observation of oocyte maturity. At present, the quality and maturity of oocytes are mainly roughly evaluated based on the size of OCC, the dispersion degree of the corona radiata and cumulus cells, or the granular cells are removed to directly observe the oocyte maturity. However, studies have shown that: the general morphology of OCC cannot directly and accurately reflect the maturity and quality of oocytes, and the misjudgment rate is relatively high; when culturing immature oocytes in vitro, it is found that granular cells play an important role in the nuclear maturation, cytoplasmic maturation of oocytes and protecting their developmental potential. Premature removal of granular cells will cause abnormal fertilization of oocytes, greatly reducing the fertilization rate or increasing the polyspermy rate, and will also reduce the blastocyst formation rate and the high-quality blastocyst rate, thereby increasing the economic burden and psychological burden of patients. Therefore, there is an urgent need to find a simple observation method and device with less impact on oocytes. Summary of the Invention

[0003] The purpose of the present invention is to provide a vessel for conveniently observing the morphology of oocytes and a using method thereof in order to solve the above problems.

[0004] The present invention achieves the above purpose through the following technical solutions:

[0005] A vessel for conveniently observing the morphology of oocytes includes a circular concave hole, a long strip-shaped groove, a non-connected groove, and a circular groove. The non-connected groove and the circular groove are arranged on the outside of the circular concave hole. The long strip-shaped groove is arranged between the circular concave hole and the circular groove. The circular concave hole and the circular groove are connected through the long strip-shaped groove. The shape of the non-connected groove is circular, and the center of the non-connected groove and the center of the circular concave hole are on a straight line.

[0006] Among them, the shape of the vessel is a cylinder, the bottom diameter is 60 mm, and the height is 10 mm. The material used for the vessel is disposable polystyrene. The diameter of the circular concave hole is 5 mm and the depth is 1 mm. The width of the long strip groove is 1 mm, the depth is 1 mm, and the length is 10 mm. The diameter of the circular groove is 10 mm and the depth is 1 mm. The diameter of the unconnected groove is 10 mm and the depth is 4 mm. The circular concave hole, the long strip groove, the unconnected groove, and the circular groove all have flat bottoms.

[0007] Among them, the number of the long strip grooves and the circular grooves is the same and is seven, and the number of the unconnected grooves is two.

[0008] The present invention also provides a method for using the vessel for conveniently observing the morphology of oocytes. When in use, first add G-IVF PLUS culture solution into the circular concave hole, the circular groove, and the unconnected groove, so that the G-IVF PLUS culture solution is paved flat at the bottoms of the circular concave hole, the circular groove, and the unconnected groove, and fills the connected long strip groove. Cover with heated mineral oil. Then add the OCC to be observed into the peripheral circular groove. Suck out the culture solution from the circular concave hole with a Pasteur pipette. Utilize the principle of communicating vessels, so that the culture solution in the circular groove connected to the circular concave hole is reduced. Slightly push the OCC by using the surface tension of the liquid. The sucked-out culture solution is temporarily stored in the unconnected groove. Observe the morphology of the oocytes under a microscope. After the observation and photography are completed, add the culture solution again, and transfer the OCC back to the vessel containing G-IVF PLUS culture solution for subsequent operations.

[0009] The advantages of the present invention are as follows:

[0010] 1. The present invention mainly uses the surface tension at the interface of the culture solution and the mineral oil to slightly squeeze the OCC, push the granulosa cells to the periphery of the oocytes, and a relatively complete morphology of the oocytes can be observed. In particular, the GV-stage oocytes can be clearly distinguished.

[0011] 2. A simple method for observing the maturity of oocytes without removing granulosa cells is proposed for the first time. Description of the Drawings

[0012] Figure 1 It is a schematic structural diagram of the vessel for conveniently observing the morphology of oocytes according to the present invention.

[0013] The description of the reference numerals in the drawings is as follows:

[0014] 1. Circular concave hole; 2. Long strip groove; 3. Unconnected groove; 4. Circular groove. Detailed Embodiments

[0015] The present invention will be further described below in conjunction with specific embodiments, but the present invention is not limited to these embodiments.

[0016] Embodiment

[0017] As Figure 1 shown, a vessel for conveniently observing the morphology of oocytes includes a circular concave hole 1, a long strip-shaped groove 2, a non-connected groove 3, and a circular groove 4. The diameter of the circular concave hole 1 is 5 mm and the depth is 1 mm. A non-connected groove 3 and a circular groove 4 are arranged outside the circular concave hole 1. The diameter of the circular groove 4 is 10 mm and the depth is 1 mm. The diameter of the non-connected groove 3 is 10 mm and the depth is 4 mm. A long strip-shaped groove 2 is arranged between the circular concave hole 1 and the circular groove 4. The width of the long strip-shaped groove 2 is 1 mm, the depth is 1 mm, and the length is 10 mm. The circular concave hole 1 and the circular groove 4 are connected through the long strip-shaped groove 2. The number of the long strip-shaped grooves 2 and the circular grooves 4 is the same and is seven. The number of the non-connected grooves 3 is two. The shape of the non-connected groove 3 is circular. The center of the non-connected groove 3 and the center of the circular concave hole 1 are on a straight line. The shape of the vessel is a cylinder, the bottom diameter is 60 mm, and the height is 10 mm. The material used for the vessel is disposable polystyrene. The circular concave hole 1, the long strip-shaped groove 2, the non-connected groove 3, and the circular groove 4 are all flat-bottomed.

[0018] The present invention also provides a method for using a vessel for conveniently observing the morphology of oocytes. When in use, first add G-IVF PLUS culture medium to the circular concave hole 1, the circular groove 4, and the non-connected groove 3, so that the G-IVF PLUS culture medium is paved flat at the bottom of the circular concave hole 1, the circular groove 4, and the non-connected groove 3, and fills the seven connected long strip-shaped grooves 2. Cover with heated mineral oil. Then add the OCC to be observed to the seven peripheral circular grooves 4. Suck out the culture medium from the circular concave hole 1 with a Pasteur pipette. Using the principle of communicating vessels, the culture medium in the seven circular grooves 4 connected to the circular concave hole 1 is reduced. Slightly push the OCC using the surface tension of the liquid. The sucked-out culture medium is temporarily stored in the two non-connected grooves 3. Observe the morphology of the oocytes under a microscope. After observing and taking pictures, add the culture medium again, and transfer the OCC back to the vessel containing G-IVF PLUS culture medium for subsequent operations.

[0019] The above-described embodiments only represent the implementation manners of the present invention. The description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the patent of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention.

Claims

1. A method of using a vessel for conveniently observing the morphology of oocytes, characterized in that: The vessel includes a circular concave hole (1), a long strip-shaped groove (2), a non-connected groove (3), and a circular groove (4). The non-connected groove (3) and the circular groove (4) are arranged outside the circular concave hole (1). The long strip-shaped groove (2) is arranged between the circular concave hole (1) and the circular groove (4). The circular concave hole (1) and the circular groove (4) are connected through the long strip-shaped groove (2). The shape of the non-connected groove (3) is circular, and the center of the non-connected groove (3) and the center of the circular concave hole (1) are on a straight line. The shape of the vessel is a cylinder, with a bottom diameter of 60 mm and a height of 10 mm. The material used for the vessel is disposable polystyrene. The diameter of the circular concave hole (1) is 5 mm and the depth is 1 mm. The width of the long strip-shaped groove (2) is 1 mm, the depth is 1 mm, and the length is 10 mm. The diameter of the circular groove (4) is 10 mm and the depth is 1 mm. The diameter of the non-connected groove (3) is 10 mm and the depth is 4 mm. The circular concave hole (1), the long strip-shaped groove (2), the non-connected groove (3), and the circular groove (4) are all flat-bottomed. The number of the long strip-shaped grooves (2) and the circular grooves (4) is the same and is seven, and the number of the non-connected grooves (3) is two; During use, first add G-IVF PLUS culture medium to the circular concave hole (1), the circular groove (4), and the non-connected groove (3), so that the G-IVF PLUS culture medium is paved flat at the bottom of the circular concave hole (1), the circular groove (4), and the non-connected groove (3), and fills the connected long strip-shaped groove (2). Cover it with heated mineral oil. Then add the OCC to be observed to the surrounding circular grooves (4). Use a Pasteur pipette to suck out the culture medium from the circular concave hole (1). Using the principle of communicating vessels, the culture medium in the circular groove (4) connected to the circular concave hole (1) is reduced. Slightly push the OCC using the surface tension of the liquid. The sucked-out culture medium is temporarily stored in the non-connected groove (3). Observe the morphology of the oocytes under a microscope. After observing and taking pictures, add the culture medium again, and transfer the OCC back to the vessel containing G-IVF PLUS culture medium for subsequent operations.

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