CN113249204B - A special device and method for separating adhesions of egg crown-cumulus complex under stereomicroscope - Google Patents

Abstract

A special device for separating adhesions of an egg corona-cumulus complex under a stereoscope comprises a main body and a replaceable suction head. The main body comprises a tube, a connecting tube is provided at the lower end of the tube, a piston rod is inserted at the upper end, a piston is provided at the lower end of the piston rod, a reset spring is provided between the piston and the tube, and a replaceable suction head is inserted in the connecting tube; the lower end of the replaceable suction head is provided with a front bevel and a rear bevel both inclined backward and upward, the outer edges of the front bevel and the rear bevel form a front bevel cutting edge and a rear bevel cutting edge, and the intersection of the front bevel and the rear bevel forms a fulcrum, and the replaceable suction head can be tilted forward and backward around the fulcrum to switch between the front bevel cutting edge and the rear bevel cutting edge. The present invention also provides a separation method of a special device for separating adhesions of an egg corona-cumulus complex under a stereoscope. The present invention can complete the complete separation of adhesions of the egg corona-cumulus complex by single-handed operation, shorten the in vitro operation time of the egg corona-cumulus complex, reduce the in vitro exposure time of oocytes, and ensure the quality of oocytes.

Description

A special device and method for separating adhesions of egg crown-cumulus complex under stereomicroscope

Technical Field

The present invention relates to the field of assisted reproductive technology, and in particular to a special device and method for separating adhesions of an egg crown-cumulus complex under a stereoscope.

Background technique

Assisted reproductive technology (ART) has become the most effective treatment for infertility patients. In the process of ART treatment, the collection and processing of the ovarian corona-cumulus complex is an important step. Under the guidance of vaginal ultrasound, the ovarian follicles are punctured, and the follicular fluid is aspirated out under negative pressure. The ovarian corona-cumulus complex is quickly detected and collected from the follicular fluid under a stereoscope, rinsed in vitro and separated from the adhesions, and then transferred to the culture medium for culture.

The human cumulus complex is composed of oocytes and outer granulosa cells. The outer granulosa cells mainly consist of two groups of cells: one group is the granulosa cells tightly wrapped around the zona pellucida of the oocyte, called the corona radiata cells; the other group of cells is a large group of cells surrounding the outer layer of the corona radiata cells, called cumulus cells. The cumulus cells are densely packed, and there is a lot of gelatinous and sticky extracellular matrix between the cells.

In the follicular fluid aspirated under negative pressure, the cumulus complex can be identified under a stereoscope as a grayish-white flaky or cloudy sticky cell mass, and the outer cumulus cells are often adhered with blood clots (Figure 6A) and large pieces of degenerated black tissue cells (Figure 6B), which cannot be removed by repeated rinsing or blowing, and the proportion is as high as 21% to 40%. Literature studies have shown that these blood clots and degenerated tissue cells may change the composition and pH value of the culture medium, hinder the release of hyaluronidase by sperm, affect sperm-egg fertilization, and thus affect the fertilization rate and embryo development potential.

At present, the separation of the adhesions of the egg crown-cumulus complex is routinely handled with a 1 ml syringe. Under a stereoscope, hold a syringe in the left hand, fix the adhesions of the egg crown-cumulus complex with the syringe needle, hold another syringe in the right hand, and use the friction between the two needles to cut and separate.

The existing adhesion separation technology has the disadvantages of inconvenient operation and low efficiency. This leads to long operation time, increased exposure time of oocytes in vitro, and fluctuations in temperature, osmotic pressure and pH value. The main reasons are: on the one hand, the structure of the 1 ml syringe itself is not conducive to operation. The needle tip gradually becomes thinner and has a long bevel with an angle of 15 to 18 degrees to the axial horizontal plane. When the adhesion is fixed with the needle tip, the contact point is very small and the interface of one separation and cutting is very limited. It is often impossible to complete the separation in one time. The needle has to be repeatedly fixed and repeatedly operated to completely separate the adhesion; on the other hand, the outer layer of the cumulus cells of the ovarian corona complex is dense, and there is a large amount of gelatinous and viscous extracellular matrix between the cells, which makes it difficult to separate the adhesion.

At present, there is no special device for separating the adhesion of the ovum complex under a stereoscope in assisted reproduction laboratories. Therefore, it is urgent to invent a special device for separating the adhesion of the ovum complex.

Summary of the invention

In order to overcome the above problems, the present invention provides a special device and method for separating adhesions of the egg crown-cumulus complex under a stereoscope.

The first aspect of the present invention provides a special device for separating adhesions of an egg-crown complex under a stereoscopic microscope, comprising a body (1) and a replaceable suction head, wherein the body comprises a tube (205), the lower end of the tube (205) is provided with a downwardly protruding connecting tube (207), and the connecting tube (207) is inserted with a replaceable suction head; the upper end of the tube (205) is provided with a piston rod (202), the upper end of the piston rod (202) is provided with a button (201), the lower end of the piston rod (202) is provided with a piston (204), and the piston (204) is slidably and sealingly fitted in the tube (205); a return spring (206) is provided between the piston (204) and the tube (205), and the two ends of the return spring (206) respectively abut against the piston (204) and the inner wall of the lower end of the tube (205);

The replaceable suction head is in the shape of a hollow cylinder. The lower end of the replaceable suction head is provided with a front bevel (303) and a rear bevel (304) both inclined backward and upward, and a mark for distinguishing the front bevel (303) and the rear bevel (304) is provided on the replaceable suction head; the outer edges of the front bevel (303) and the rear bevel (304) respectively form a front bevel cutting edge and a rear bevel cutting edge, and the intersection of the front bevel (303) and the rear bevel (304) forms a fulcrum (305), and the replaceable suction head can be tilted forward and backward around the fulcrum to switch between using the front bevel cutting edge and the rear bevel cutting edge; the upper end of the replaceable suction head is provided with a filter membrane layer (301), the filter membrane layer (301) closes the upper end opening of the replaceable suction head, and the filter membrane layer (301) is a microporous filter membrane; after the return spring (206) is fully compressed, the volume of air discharged from the tube (205) is equal to the volume of the replaceable suction head.

Furthermore, the horizontal length L1 of the front inclined surface (303) is greater than the horizontal length L2 of the rear inclined surface (304), and the angle α between the front inclined surface (303) and the horizontal plane is smaller than the angle β between the rear inclined surface (304) and the horizontal plane.

Furthermore, the angle α between the front bevel (303) and the horizontal plane is 15 to 25 degrees, and the horizontal length L1 is 50 to 75% of the inner diameter of the suction head; the angle β between the rear bevel (304) and the horizontal plane is 35 to 45 degrees, and the horizontal length L2 is 25 to 50% of the inner diameter of the suction head.

Furthermore, the vertical axis length L3 of the front bevel cutting edge and the rear bevel cutting edge is 2 to 3 mm, and the angle δ of the front bevel cutting edge and the rear bevel cutting edge is 30 to 45 degrees.

Furthermore, a scale (302) is provided on the upper part of the replaceable suction tip, and the scale (302) is used to indicate the amount of liquid sucked into the suction tip, and a maximum liquid suction amount warning line is marked on the scale (302); and a protective cover is provided on the outer cover of the replaceable suction tip.

Furthermore, a tube wing (203) is provided on the outer wall of the rear end of the tube (205), and the tube wing (203) has a first wing edge and a second wing edge corresponding to the index finger and the middle finger, the first wing edge is a downwardly curved arc structure, and the second wing edge is a straight line structure.

Furthermore, the button (201) is circular or elliptical, and the top surface of the button (201) has a recessed portion, the area of which matches the area of the thumb.

Furthermore, the inner diameter of the tube (205) is 8 to 10 mm, and the length of the tube (205) is 80 to 100 mm.

Furthermore, the pore size of the microporous filter membrane is 0.22 microns.

The second aspect of the present invention provides a method for separating adhesions of the egg crown-cumulus complex under a stereoscope using the special device, comprising the following steps:

Step 1, connect the main body and the replaceable suction head in the front and back directions as marked;

Step 2, find the egg crown cumulus complex from the follicular fluid under a stereoscope, press the button, the reset spring is compressed, the air is squeezed and discharged from the lower end of the suction head, the suction head is immersed in the follicular fluid containing the egg crown cumulus complex, the button is slowly released, the piston moves up, and the reset spring also slowly resumes its extension, and the egg crown cumulus complex is sucked from the follicular fluid into the suction head; in the cleaning fluid, the button is pressed down to discharge the egg crown cumulus complex in the suction head into the cleaning fluid, completing the transfer of the egg crown cumulus complex from the follicular fluid to the cleaning fluid;

Step 3, using the special tool to separate the adhered matter in the cleaning solution according to the following "three-step method":

In the first step, a part of the oocyte in the cumulus-ovum complex is sucked out with a pipette tip, and the adherent material remains outside the pipette tip;

The second step is to press down the posterior bevel forcefully and drag it backwards by 1 to 3 mm. The pressure between the posterior bevel edge and the bottom of the dish causes the cumulus cell cluster near the posterior bevel edge to be subjected to the combined effects of shear stress and tensile stress, causing local thinning. Then, the cumulus cells around the adhesion are squeezed forward to the anterior bevel by moving the relative distance between the posterior bevel edge and the bottom of the dish.

The third step is to use the intersection of the front bevel and the back bevel as a fulcrum, tilt forward, and switch to the front bevel edge; press down the front bevel with force, and drag it back 3 to 5 mm; through the pressure of the front bevel edge of the suction head and the bottom of the dish, the cumulus cell clusters near the front bevel edge are simultaneously subjected to the combined effects of shear stress and tensile stress, and the cell clusters on both sides of the front bevel edge become thinner; then, through the relatively long-distance movement of the front bevel edge and the bottom of the dish, the shear stress and tensile stress on the cumulus cells on both sides of the front bevel edge are further increased, and at the same time, the adhesion between cells is further destroyed, thereby producing a separation section, and completely completing the separation of the cumulus complex and the adhesion;

Step 4, refer to step 2 and transfer the egg crown-cumulus complex from the cleaning solution to the culture solution after the adhesion is separated.

The beneficial effects of the present invention are:

(1) One-hand operation, simplified steps. By using the special device for separating the adhesions of the egg-corona-cumulus complex of the present invention, laboratory personnel can complete the complete separation of the adhesions of the egg-corona-cumulus complex with one hand operation, and at the same time complete the transfer of the egg-corona-cumulus complex without changing the instrument, thus overcoming the disadvantage of repeatedly changing the instrument in the traditional operation process and greatly simplifying the operation steps.

(2) Shortening the operation time. The special device of the present invention can achieve two functions, namely, while separating the adhesion of the ovarian corona-cumulus complex, it can also realize the transfer of the ovarian corona-cumulus complex from follicular fluid-cleaning fluid-culture fluid, greatly shortening the in vitro operation time of the ovarian corona-cumulus complex.

(3) Ensure the quality of oocytes. The simple operation and simplified steps reduce the exposure time of oocytes in vitro and avoid the decline of oocyte quality caused by fluctuations in environmental temperature, osmotic pressure and pH value.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is a flow chart of the transfer of the egg crown-cumulus complex and the separation of its adherents.

FIG. 2 is a schematic diagram of the structure of the main body.

FIG. 3 is a side view of an interchangeable nozzle.

FIG. 4 is a rear view of the replaceable suction tip.

FIG. 5 is a schematic structural diagram of the front bevel edge and the rear bevel edge of the replaceable suction head.

FIG. 6A is a picture showing that the adherent substance of the oocyte-corona complex is a blood clot (the solid arrow indicates the oocyte, and the hollow arrow indicates the blood clot).

FIG6B is a picture showing that the adherents of the oocyte-corona complex are degenerated tissue cells (solid arrows indicate oocytes, and hollow arrows indicate degenerated tissue cells).

Description of reference numerals: 1-body; 201-button; 202-piston rod; 203-tube wing; 204-piston; 205-tube; 206-reset spring; 207-connecting tube; 301-filter membrane layer; 302-scale; 303-front bevel; 304-rear bevel; 305-fulcrum. L1-horizontal length of the front bevel; L2-horizontal length of the rear bevel; L3-vertical axis length of the cutting edge; α-angle between the front bevel and the horizontal plane; β-angle between the rear bevel and the horizontal plane; δ-angle of the cutting edge away from the axis of the suction head.

Detailed ways

The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

In the description of the present invention, it should be noted that the orientations or positional relationships indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. are based on the orientations or positional relationships shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", and "third" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance.

In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected" and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

Embodiment 1: A special device for separating adhesions of the egg crown-cumulus complex under a stereoscope

The structure of the special device for transferring the egg crown-cumulus complex and separating the adhesions under a stereoscope of the present invention is as follows:

As shown in FIG. 2 , the device comprises a body 1 and a replaceable suction head. The body comprises a tube 205, the tube 205 has an inner diameter of 10 mm and a length of 80 mm, a downwardly protruding connecting tube 207 is provided at the lower end of the tube 205, and a replaceable suction head is inserted in the connecting tube 207; a piston rod 202 is inserted at the upper end of the tube 205, a button 201 is provided at the upper end of the piston rod 202, a piston 204 is provided at the lower end of the piston rod 202, and the piston 204 is slidably and sealingly fitted in the tube 205; a return spring 206 is provided between the piston 204 and the tube 205, and the two ends of the return spring 206 respectively abut against the piston 204 and the inner wall of the lower end of the tube 205; the volume of air discharged after the return spring 206 is fully compressed should be equal to the volume of the suction head.

The outer wall of the rear end of the tube 205 is provided with a tube wing 203, which has a first wing side and a second wing side corresponding to the index finger and the middle finger, the first wing side is a downward curved structure, the second wing side is a straight structure, the curved structure is consistent with the second joint of the middle finger to reduce the pressure of the thumb during the pressing operation, and the arrangement of the curved structure is convenient for the main body 1 to be hung when not in use. The button 201 is circular or oval, and the top surface of the button 201 has a concave part, the area of which is equivalent to the area of the thumb, which is conducive to improving the stability of the hand during operation and relieving muscle fatigue in the thumb area.

As shown in FIG. 3 to FIG. 5 , the replaceable suction head is in the shape of a hollow cylinder, and a front inclined surface 303 and a rear inclined surface 304 both inclined upward and backward are provided at the lower end of the replaceable suction head, and a color mark is provided at the lower end of the suction head to distinguish the front and the back. The outer edges of the front bevel 303 and the rear bevel 304 are gradually thinned to form a front bevel edge and a rear bevel edge respectively; the intersection of the front bevel 303 and the rear bevel 304 forms a fulcrum 305, and the replaceable tip can be tilted forward and backward around the fulcrum to switch between the front bevel edge and the rear bevel edge; a filter membrane layer 301 is provided at the upper end of the replaceable tip, and the filter membrane layer 301 closes the upper end opening of the replaceable tip, and the filter membrane layer 301 is a microporous filter membrane with a pore size of 0.22 microns; the lower part of the tip is made of metal and the upper part is made of transparent plastic; a scale 302 indicating the amount of inhaled liquid is provided at the upper part of the tip, which has three scales: 300μl, 400μl, and 500μl, and a warning line of 500μl is specially marked for the maximum amount of liquid inhaled, so as to avoid the filter membrane layer from being blocked and contaminated due to the inhaled liquid exceeding the maximum amount of liquid entering the filter membrane layer. After the return spring 206 is fully compressed, the volume of air discharged from the tube 205 is equal to the volume of the replaceable suction tip;

In this embodiment, the replaceable suction head has an inner diameter of 4 mm, a length of 55 mm, and is provided with a protective cover. The lower part of the suction head is made of metal material, and the upper part is made of hard transparent plastic material, and the connection between the metal material and the plastic material is made of plastic hot riveting technology.

In this embodiment, the horizontal angle α of the front bevel 303 of the suction head is 15 to 25 degrees, and the horizontal angle β of the rear bevel 304 is 35 to 45 degrees. The horizontal length L1 of the front bevel 303 of the suction head is 2.5 mm, and the horizontal length L2 of the rear bevel is 1.5 mm. The cutting edges of the front bevel 303 and the rear bevel 304 can be completed by mechanical fine grinding or laser heat treatment, and the vertical axis length L3 of the cutting edge is 3 mm. The cutting edge angle deviates from the axis of the suction head, and the angle δ is 30 degrees.

Example 2: Test of the present invention for separating the adhesion of the ovum complex under a stereoscope

The tested tips are: No. 1: the angle α between the front bevel of the tip and the horizontal plane is 25 degrees, and the horizontal angle β of the rear bevel is 45 degrees; No. 2: the horizontal angle α of the front bevel is 25 degrees, and the horizontal angle β of the rear bevel is 35 degrees; No. 3: the horizontal angle α of the front bevel is 20 degrees, and the horizontal angle β of the rear bevel is 45 degrees; No. 4: the horizontal angle α of the front bevel of the tip is 20 degrees, and the horizontal angle β of the rear bevel is 35 degrees; No. 5: the horizontal angle α of the front bevel is 15 degrees, and the horizontal angle β of the rear bevel is 45 degrees; No. 6: the horizontal angle α of the front bevel is 15 degrees, and the horizontal angle β of the rear bevel is 35 degrees.

The test method steps are described as follows:

1. Collect the discarded granular cell clusters, transfer them to a culture dish with culture medium, and mark them with numbers. Take the pipette tip to be tested, connect them to the body in Example 1 in the front and back directions of the mark, and remove the protective cover of the pipette tip.

2. Operate according to the "three-step method" of the present invention under a stereoscope.

The first step is to press the button to expel the air, immerse the tip in the culture medium, slowly release the button, and suck half of the granular cell cluster into the tip, leaving the other half outside the tip.

The second step is to press down the back bevel hard so that the edge of the back bevel is close to the bottom of the dish, and drag it backwards 1 to 3 mm to squeeze all the granular cells remaining outside the pipette tip forward to the front bevel.

Step 3: Use the intersection of the front bevel and the back bevel as a fulcrum, tilt forward, and switch to the front bevel edge. Press down the front bevel hard to make the front bevel edge close to the bottom of the dish, and drag it back 3 to 5 mm to complete the complete separation of the granular cells inside and outside the pipette tip. If the separation is not complete at one time, repeat the second and third steps above.

3. Move the granular cell cluster in the pipette tip to another culture dish filled with culture medium, press the button to discharge the granular cell cluster in the pipette tip into the culture medium.

4. Observe whether the cutting and separation are complete under the stereoscope. Repeat the above operation 10 times for each group, and use a specially designed table to record the test results. The judgment criteria for the separation and cutting of granular cell clusters: ① The granular cell clusters are completely separated at one time; ② The granular cell clusters can be completely separated, but it takes more than or equal to two times to be completely separated; ③ The granular cell clusters cannot be completely separated, that is, the separation fails. For example, if "the granular cell clusters can be completely separated at one time" appears once in 10 separation operations, it is 10%.

Table 1 Detection results of adhesions of the egg crown-cumulus complex separated by different numbered tips

The test results are shown in Table 1. It can be seen from the test results that six specifications of pipette tips can completely separate the particle cell clusters, but the separation effect of No. 1 pipette tip is the best, 80% can be completely separated in one time, and only 20% can be completely separated after multiple cuts. Only 50% of the No. 5 pipette tip can be completely separated in one time, and 50% can be completely separated after multiple cuts. 40% of the No. 3 pipette tip needs multiple cuts to be completely separated, and 60% can be completely separated in one time. The reason for this may be that the front bevel of No. 1 pipette tip is longer than that of No. 3, No. 4, No. 5 and No. 6 pipette tips, with a larger cutting range, and the 45-degree angle of the rear bevel is more conducive to operation, so the separation is more thorough.

Example 3: Comparison of the operation time of operators with different proficiency levels using the apparatus of the present invention and conventional methods to separate the adhesion of the ovum complex

Operators with different proficiency levels used the apparatus of the present invention and the conventional method (i.e., syringe separation method) to separate the adhesions of the egg crown-cumulus complex under a stereoscope. The different proficiency levels refer to proficiency, that is, having more than 300 cycles of egg crown-cumulus complex collection experience; basic mastery, that is, having 100 to 300 cycles of egg crown-cumulus complex collection experience; and beginner, that is, less than 100 cycles of egg crown-cumulus complex collection experience. Each group was repeated 10 times. Syringe separation method: Use two 1 ml syringes, hold one syringe in the left hand, fix the adhesions with the syringe needle, hold the other syringe in the right hand, and use the friction between the needles to cut and separate. The separated granular cell clusters are then transferred to a new culture dish filled with culture medium using a Pasteur pipette. Separation method of the present invention device: operate according to the methods of steps 2 and 3 in Example 2. Timing is from the beginning of separation of granular cell clusters to transfer to a new culture dish.

Table 2 Comparison of operation time of two methods for separating adhesions of the ovalbumin complex by different operators

Note: ^a : relative to the syringe separation method, P<0.05; ^b : relative to the syringe separation method, P<0.05; ^c : relative to the syringe separation method, P<0.05; ^e : relative to beginners, P<0.05; ^f : relative to those who have mastered the basic skills, P<0.05.

The results show that: 1. The operation time of the same operator using the special device of the present invention to separate the adhesions of the egg corona and cumulus complex is significantly lower than that of the conventional syringe separation method. 2. The operation time of the skilled operator using the syringe separation method to separate the adhesions of the egg corona and cumulus complex is significantly less than that of the beginner and the basic master, but there is no significant difference in the operation time of the beginner, the basic master and the skilled operator using the special device of the present invention to separate the adhesions, indicating that the use method of the special device of the present invention is simple and easy to learn, and can be used without long-term training. Conventional use of a syringe to separate the adhesions of the egg corona and cumulus complex often requires multiple separations, and when the egg corona and cumulus complex is transferred after separation, the instrument needs to be replaced, and this process is cumbersome and time-consuming. The special device of the present invention can complete the separation of the adhesions of the egg corona and cumulus complex with one hand, and the transfer of the egg corona and cumulus complex in the next step can be completed without changing the instrument, which greatly simplifies the operation steps, shortens the exposure time of the oocyte in vitro, avoids the decline in oocyte quality caused by the drop in ambient temperature, osmotic pressure and pH fluctuations, and significantly improves work efficiency.

The contents described in the embodiments of this specification are merely an enumeration of the implementation forms of the inventive concept. The protection scope of the present invention should not be regarded as limited to the specific forms described in the embodiments. The protection scope of the present invention also extends to equivalent technical means that can be conceived by those skilled in the art based on the inventive concept.

Claims (7)

1. An apparatus special for separating the adherends of a crown-cumulus compound under a body view mirror is characterized in that: the novel suction head comprises a body (1) and a replaceable suction head, wherein the body comprises a pipe barrel (205), a connecting pipe (207) protruding downwards is arranged at the lower end of the pipe barrel (205), and the replaceable suction head is inserted into the connecting pipe (207); a piston rod (202) is inserted into the upper end of the pipe barrel (205), a button (201) is arranged at the upper end of the piston rod (202), a piston (204) is arranged at the lower end of the piston rod (202), and the piston (204) is matched in the pipe barrel (205) in a sliding and sealing manner; a return spring (206) is arranged between the piston (204) and the tube (205), and two ends of the return spring (206) are respectively abutted against the inner walls of the lower ends of the piston (204) and the tube (205);

The replaceable suction head is hollow cylindrical, a front inclined surface (303) and a rear inclined surface (304) which are inclined upwards and backwards are arranged at the lower end of the replaceable suction head, and marks for distinguishing the front inclined surface (303) from the rear inclined surface (304) are arranged on the replaceable suction head; the outer edges of the front inclined plane (303) and the rear inclined plane (304) respectively form a front inclined plane cutting edge and a rear inclined plane cutting edge, a fulcrum (305) is formed at the junction of the front inclined plane (303) and the rear inclined plane (304), and the replaceable suction head can incline forwards and backwards around the fulcrum so as to switch between the front inclined plane cutting edge and the rear inclined plane cutting edge; the upper end of the replaceable suction head is provided with a filter membrane layer (301), the filter membrane layer (301) seals the upper end opening of the replaceable suction head, and the filter membrane layer (301) is a microporous filter membrane; the volume of air discharged by the tube (205) after the reset spring (206) is fully compressed is equal to the volume of the replaceable suction head;

The horizontal length L1 of the front inclined plane (303) is larger than the horizontal length L2 of the rear inclined plane (304), and the included angle alpha of the front inclined plane (303) and the horizontal plane is smaller than the included angle beta of the rear inclined plane (304) and the horizontal plane; the included angle alpha between the front inclined surface (303) and the horizontal plane is 15-25 degrees, and the horizontal length L1 is 50-75% of the inner diameter of the suction head; the included angle beta between the rear inclined plane (304) and the horizontal plane is 35-45 degrees, and the horizontal length L2 is 25-50% of the inner diameter of the suction head;

the vertical axial length L3 of the front inclined plane cutting edge and the rear inclined plane cutting edge is 2-3 mm, and the angle delta of the front inclined plane cutting edge and the rear inclined plane cutting edge is 30-45 degrees.

2. The apparatus for separating the adherent of a compound of a crown and a cumulus under a body view mirror according to claim 1, wherein: the upper part of the replaceable suction head is provided with a graduated scale (302), the graduated scale (302) is used for representing the liquid amount sucked into the suction head, and the graduated scale (302) is marked with a warning line of the maximum liquid suction amount; the replaceable suction head is sleeved with a protective sleeve.

3. The apparatus for separating the adherent of a compound of a crown and a cumulus under a body view mirror according to claim 1, wherein: the rear end outer wall of the tube (205) is provided with a tube wing (203), the tube wing (203) is provided with a first wing edge and a second wing edge which correspond to the index finger and the middle finger, the first wing edge is of a downward bending arc-shaped structure, and the second wing edge is of a straight-line-shaped structure.

4. A tool for separating the adherent of a stereoscopic sub-cumulus complex according to claim 3, wherein: the button (201) is round or oval, the top surface of the button (201) is provided with a concave part, and the area of the concave part is matched with the area of the thumb.

5. The apparatus for separating the adherent of a compound of a crown and a cumulus under a body view mirror according to claim 1, wherein: the inner diameter of the pipe barrel (205) is 8-10 mm, and the length of the pipe barrel (205) is 80-100 mm.

6. The apparatus for separating the adherent of a compound of a crown and a cumulus under a body view mirror according to claim 1, wherein: the pore size of the microporous filter membrane is 0.22 micron.

7. A method for the separation of the adherent of a stereoscopic cumulus complex using a special instrument according to any one of claims 1-6, comprising the steps of:

step 1, connecting a body and a replaceable suction head according to the front-back direction of a mark;

step 2, searching a follicle-hillock compound from follicular fluid under a body view mirror, pressing down a button, compressing a reset spring, extruding air and discharging the air from the lower end of a suction head, immersing the suction head into the follicular fluid containing the follicle-hillock compound, slowly releasing the button, moving up the piston, and slowly recovering the reset spring to stretch along with the button, so that the follicle-hillock compound is sucked into the suction head from the follicular fluid; pressing down a button in the cleaning liquid to discharge the cumulus compound in the suction head into the cleaning liquid to finish the transfer of the cumulus compound from the follicular liquid to the cleaning liquid;

step 3, separating the adhesion substances in the cleaning solution by using the special tool according to the following three-step method:

Firstly, sucking part of oocyte in the cumulus complex of the ovum by using a suction head, and keeping the adhesion outside the suction head;

secondly, pressing down the rear inclined plane with force and dragging backwards for 1-3 mm; the cumulus cells near the rear inclined plane cutting edge are subjected to the combined action of shear stress and tensile stress by utilizing the compression of the rear inclined plane cutting edge and the bottom of the dish, are locally thinned, and then the cumulus cells around the adhesion object are extruded forwards to the front inclined plane through the movement of the rear inclined plane cutting edge and the relative distance of the bottom of the dish;

Thirdly, taking the junction of the front inclined plane and the rear inclined plane as a fulcrum, tilting forwards, and switching to use the front inclined plane cutting edge; pressing the front inclined plane with force and dragging backwards for 3-5 mm; the cumulus cell mass near the front inclined plane cutting edge is subjected to the combined action of shear stress and tensile stress simultaneously by the compression of the front inclined plane cutting edge of the suction head and the bottom of the dish, and the cell mass at the two sides of the front inclined plane cutting edge is thinned; then, by moving the front bevel edge and the bottom of the dish relatively long distance, the shearing stress and the tensile stress borne by the cumulus cells at the two sides of the front bevel edge are further increased, and meanwhile, the adhesion force among the cells is further destroyed, so that a separation section is generated, and the separation of the cumulus compound and the adhesive is thoroughly completed;

And 4, referring to the step 2, transferring the cumulus complexes after the adhesion object separation from the cleaning solution to the culture solution.