33 results about "Polar body" patented technology

Disclosed is a method for using a first polar body and a second polar body as well as a single-cell embryo to carry out whole-genome non-exponential amplification and high-throughput genome sequencing, so as to perform preimplantation genetic diagnosis for genetic disease and testing for pathogenic genes causing repeated miscarriages. The method of the present invention comprises the following steps: (1) obtaining oocytes and embryos, and carrying out separation and genome amplification of first and second polar bodies and single-cell embryos; (2) establishing a genome sequencing library and sequencing, and carrying out bioinformatic analysis of the genome to obtain a gene spectrum and information concerning the number of copies of chromosomes and fragments thereof; (3) determining the chromosome ploidy of the polar bodies and embryos as well as information concerning defects, replication and point mutation in the chromosome fragments; (4) selecting normal or suitable embryos for implantation.

The invention discloses a porcine oocyte in vitro maturation medium and methods of preparation and culture. The medium comprises a TCM-199 base medium and the added porcine follicular fluid, fetal bovine serum, eCG, hCG, EGF and IGF-1. The porcine oocyte in vitro maturation medium of the invention can significantly increase the rate of maturation of porcine oocytes in vitro. Experiments confirm that after the oocyte collection and 42-44h maturation, the medium of the invention can have a polar body rate stabilized at 80% and a parthenogenetic embryo development rate up to 85%, and the entire maturation process does not require replacement of the medium and is easy to operate.

The invention discloses a time point quantification treatment method for producing a Crassostrea hongkongensis all-triploid. The time point quantification treatment method in which a fertilized egg development stage is taken as a biological indicator is created through the technical links of collection of single fertilized eggs, timely treatment of the fertilized eggs, detection of larva ploidy and the like, so that a 100-percent Crassostrea hongkongensis all-triploid can be obtained. Through optimization and pairing of the single fertilized eggs, the defect of poor mixed fertilized egg synchronicity in a conventional Crassostrea hongkongensis triploid induction process is overcome effectively, and zygotes with high synchronicity are obtained. A conventional fixed thinking mode in which a time point when 30-50 percent of a first polar body occurs is taken as a treatment time point and the optimal time interval is 20 minutes is overturned. Instead, the fertilized egg development stage is taken as the biological indicator, and A+1 / 3B and B+C are taken as a treatment time point and a treatment time interval respectively, so that the influences of external environmental conditions such as temperature on treatment are avoided, and the Crassostrea hongkongensis all-triploid can be induced successfully.

The present invention relates to a compact motorized rotational stage for microscopy applications and control methods for automated sample orientation / rotation. The rotational stage includes a motor, a rotational motion transmission mechanism, and a rotating sample holder for accommodating a holding device such as glass slides / Petri dishes of different sizes. Mouse embryos are used as an example to explain the control methods. A pattern recognition utility was developed for identifying mouse embryo structures. The transformation between the holding device rotational coordinate frame and the translational positioning stage coordinate frame is calibrated during image-based visual servo control. The polar body of an embryo is oriented through purely image-based visual servo control or through coordinate transformation and closed-loop position control.

The invention discloses a seed making method for a low-copper Fujian oyster breeding system and relates to an aquatic organism genetic breeding technique. The method comprises the following steps: selecting individuals of tidy shell shapes from a same sexual maturity Fujian oyster colony as a basic breeding group, and carrying out temporary rearing; testing the content of copper of a part of pallium, and carrying out in-situ preservation on rest soft parts under a low-temperature high-moisture condition; testing the content of copper in the pallium by using an inductively coupled plasma mass spectrometry method; carrying out anabiosis on parental oyster, carrying out artificial fertilization, and after fertilization succeeds and polar bodies grow out, washing off excessive sperms; hatchinggerm cells of the group breeding system in a culture pond, culturing floating larvae, and continuously culturing juvenile mollusk after attachment and metamorphose of the floating larvae; transferring the cultured juvenile mollusk into a same culture sea area, culturing so as to obtain a first filial generation low-copper breeding system, wherein the first filial generation develops till sexual maturity and meets commercial specifications 9-12 months later; selecting individuals which are tidy in shell shape and similar in size from the first filial generation in sexual maturity, and repeating the operation for 1-2 times till character immobilization, thereby obtaining a novel Fujian oyster strain with low copper enrichment.

The invention discloses a method for inducing ocean seashell polyploidy by using sodium chloride. The method is characterized by comprising the following steps: selecting the seashell to grow the shell to respectively obtain sperms and ovum; carrying out artificial insemination; continuously observing the developing condition of zygotes through microscopic examination to determine the number of the water body for culturing the zygotes; adding sodium chloride into the culture sea water containing the zygotes according to a proportion that 20-40 go of sodium chloride is added in 1000ml of culture seat water when 40-50 percent of zygotes are observed to release a first polar body or a first zygote is observed to occur the first polar body, so as to prevent the release of a second polar body or the first polar body of the zygotes; evenly stirring and treating for 10-15 min; and transferring the zygotes to normal sea water to incubate . The ocean seashell comprises Pacific oyster, chlamys farreri, Argopecten irradians, Crassostrea rivularis, clam, Scapharca broughtonii and the like. The seashell zygotes are treated by using the sodium chloride and the initiating agent so as to inhibit the release of the polar body and produce the shell polyploidy with no toxicity, efficiency, simple operation and extremely low cost.

The invention discloses a production method of ICSI operation vessel. The production method comprises the following steps under the following conditions: sucking PVP solution by a suction tube, forming two 10 microliter circular micro-droplets above the ICSI vessel and forming a strip-shaped PVP micro-droplet with a length of 2.5cm and a width of 0.5cm under the ICSI vessel; sucking about 0.1ml of a culture medium containing sperms, and forming a strip-shaped micro-droplet with a length of about 2.0cm and a width of about 0.5cm at one side of the strip-shaped PVP micro-droplet; sucking a buffer culture medium, and forming three to five 10 microliter circular micro-droplets containing an ovum at the other side of the strip-shaped PVP micro-droplet; sucking the buffer culture medium, and forming one 10 microliter circular micro-droplet under the strip-shaped PVP micro-droplet; sucking one sperm from the strip-shaped micro-droplet of the culture medium containing sperms by a microscopic puncture needle, placing the sperm in the strip-shaped PVP micro-droplet, and immobilizing the sperm; fixing the ovum by a microscopic fixation needle so that the polar bodies of the ovum can be located at a six-o'clock position or a twelve-o'clock position; injecting the sperm in the ovum by the microscopic puncture needle with the sperm from a three-o'clock position.

The invention belongs to the bio-medical field, relates to a polar body genome and an application thereof, and particularly relates to a polar body genome reconstruction ovum, a preparation method and an application thereof. In the invention, according to a characteristic that a polar body not only contains female DNA being same as that in egg cell cytoplasm but also carries little even no cytoplasm substances (mitochondria), the polar body genome reconstruction ovum and an application thereof in preparation of a material for preventing mitochondria maternal genetic diseases are provided in the invention. An animal model test proves that the reconstruction ovum and progenies thereof are free of mitochondria DNA of patients, so that the reconstruction ovum can effectively stop cytoplasm-related diseases and can actively and completely prevent the mitochondria maternal genetic diseases.

The invention discloses an oocyte in-vitro maturation culture solution additive. The in-vitro maturation culture solution additive contains ceruloplasmin. Therefore, a first polar body excretion rateof oocytes can be significantly increased, the ROS content of the oocytes can be significantly reduced, and the embryonic blastocyst rate after parthenogenetic activation can be significantly increased, so that the maturation rate and quality of the oocytes can be greatly improved.

The invention relates to a computer assisted embryo selection method. Before ICSI is conducted and when short time insemination oocyte dismantling observation is conducted, Nikon NIS-Elements software is used for collecting oocyte images one by one, the thickness of a transparent tape, the diameter of oocytes and the oocyte gap of a polar body position are measured, and the oocytes with the thickness of the transparent tape being 16.9-20.7 microns, the diameter of the oocytes being 114.8-119.9 microns and the oocyte gap of the polar body position being 8.5-12.3 microns are selected for being subjected to insemination; then embryos with the high developmental potentiality are selected. According to the method, the Nikon NIS-Elements software is used for collecting the oocyte images one by one, the thickness of the transparent tape, the diameter of the oocytes and the oocyte gap of the polar body position are measured, the appropriate oocytes are selected for being subjected to insemination, and then the embryos with the high developmental potentiality are selected. According to the method, a layer of screening is added compared with the prior art, and the embryos with the high developmental potentiality can be better obtained. The data obtained by using the Nikon NIS-Elements software for measuring and analyzing are more accurate.

The invention provides a cultivation method for rapidly growing triploid crassostrea gigas Haida No.1. The method comprises the following steps: obtaining high-heterozygosity triploid crassostrea gigas Haida No.1 by inhibiting discharge of first polar bodies of fertilized eggs of diploid crassostrea gigas Haida No.1, taking females of the triploid crassostrea gigas Haida No.1 as female parents, taking males of the diploid Haida No.1 as male parents, and obtaining tetraploid crassostrea gigas Haida No.1 with improved growth characters by inhibiting discharge of the first polar bodies of the fertilized eggs; and hybridizing males of the tetraploid crassostrea gigas Haida No.1 and the diploid crassostrea gigas Haida No.1 to obtain the triploid crassostrea gigas Haida No.1 which grows quickly. According to the method, the growth performance of the crassostrea gigas is improved by adopting a method for improving the heterozygosity of parent genes, and the tetraploid crassostrea gigas Haida No.1 obtained by adopting the method is hybridized with the diploid crassostrea gigas Haida No.1 which originally has the characteristic of rapid growth to obtain the triploid crassostrea gigas Haida No.1 with rapid growth performance.

The invention provides an induction method of a high-heterozygosity tetraploid of a new variety of crassostrea gigas 'Haida No.3'. The method comprises the following steps: taking female triploid crassostrea gigas 'Haida No.3' obtained by inhibiting discharge of first polar bodies of fertilized eggs of diploid crassostrea gigas 'Haida No.3' as a female parent; taking a male diploid 'Haida No.3' as a male parent; after the female parent and the male parent are artificially matured, putting ova into seawater to be matured, and when the ova are round through microscopic examination, the ova are matured; conducting insemination on the cured ova and sperms, starting timing after the sperms and the ova are mixed, treating fertilized ova with cytochalasin B, inhibiting discharging of the first polar bodies of the fertilized ova, and collecting and soaking the fertilized ova; and transferring the fertilized ova into a cultivation container for incubation and larva cultivation. The triploid female parent shellfish is obtained by inhibiting the discharge of the first polar bodies of the fertilized ova of the diploid crassostrea gigas 'Haida No.3', and the tetraploid obtained by the triploid female parent shellfish has higher growth advantage due to higher gene heterozygosity.

The invention belongs to the technical field of genetic breeding of aquaculture varieties, and relates to a method for producing tetraploid oysters and a method for interspecific hybridization of triploid oysters by using the tetraploid oysters. The triploid oysters are stimulated for 5-8 days at the high temperature of 26-28 DEG C, ripening is promoted for 30-40 days at the water temperature of 22-24 DEG C after stimulation, and ova of female oysters with the largest diameter are selected after ripening; the selected ova and sperms of diploid oysters are fertilized for 5-8 minutes, then an inhibitor is added to inhibit first polar body discharge of the fertilized ova, and then a large number of tetraploid oysters are stably obtained for optimization. After strong optimization, high-concentration sperms are used for carrying out hybrid fertilization on excellent tetraploid oysters and excellent diploid oysters in different species, and interspecific hybrid triploid oysters with excellent breeding traits are obtained. According to the method for producing the tetraploid oysters and the method for interspecific hybridization of the triploid oysters by using the tetraploid oysters, the method for producing the tetraploid oysters and the method for interspecific hybridization of the triploid oysters by using the tetraploid oysters can be used for stably and efficiently producing the tetraploid oysters and the hybrid triploid oysters with the excellent breeding traits, can be widely applied to genetic breeding of oysters and shellfishes, and improves the shellfish breeding industry.

The invention discloses a time point quantification treatment method for producing a Crassostrea hongkongensis all-triploid. The time point quantification treatment method in which a fertilized egg development stage is taken as a biological indicator is created through the technical links of collection of single fertilized eggs, timely treatment of the fertilized eggs, detection of larva ploidy and the like, so that a 100-percent Crassostrea hongkongensis all-triploid can be obtained. Through optimization and pairing of the single fertilized eggs, the defect of poor mixed fertilized egg synchronicity in a conventional Crassostrea hongkongensis triploid induction process is overcome effectively, and zygotes with high synchronicity are obtained. A conventional fixed thinking mode in which a time point when 30-50 percent of a first polar body occurs is taken as a treatment time point and the optimal time interval is 20 minutes is overturned. Instead, the fertilized egg development stage is taken as the biological indicator, and A+1 / 3B and B+C are taken as a treatment time point and a treatment time interval respectively, so that the influences of external environmental conditions such as temperature on treatment are avoided, and the Crassostrea hongkongensis all-triploid can be induced successfully.

The invention discloses an induction method for a haliotis discus hanai triploid, and relates to a shellfish culture method. The method comprises the steps of selecting abalone seeds and promoting maturity; performing artificial spawning induction; selecting and processing gametes; performing synchronous fertilization; performing medicine induction; removing medicine; performing incubation. On thepremise of ensuring haliotis discus hanai seed gonad maturity and synchronous fertilization, the induction condition is further optimized, and therefore the haliotis discus hanai triploid is obtainedefficiently and stably. Synchronous fertilization is ensured, the phenomenon that gametes are in contact in advance in the discharge process is strictly avoided, microscopic examinations determine that no accident fertilization exists, and the consistency of synchronous fertilization and germ cell development is ensured. The induction conditions are optimized, wherein constant microscopic examination observation determines that 70-80% of contrast group germ cells PB1 are adopted as a processing time node, it is ensured that a first polar body is effectively released, and tetraploids and aneuploids are avoided. According to the stable and efficient induction rate, the induction rate of the triploid is 100% or is close to 100%.

The invention relates to a technique for promoting in vitro maturation of human immature oocytes. A CNP based biphasic in vitro maturation (CNP based biphasic IVM, C-IVM) technique for human immatureoocytes simulates an in vivo development process of oocytes to realize in vitro maturation of the oocytes in a phased manner: after immature cumulus-oocyte complexes (COCs) are obtained from small antral follicles, firstly (1) preculture (pre-IVM) is performed, meiosis of the oocytes is restrained through C-type natriuretic peptide (CNP), and besides, cytoplasm maturing of the oocytes is promoted;and (2) in vitro maturation (IVM) culture is performed, maturing of oocyte nuclei is promoted through amphiregulin (AREG) and FSH, first polar bodies are discharged, and MII oocytes are formed. Through the technique disclosed by the invention, the human oocytes low in maturity can be promoted to form mature oocytes having high development potential in vitro, the state of low IVM efficiency at present is significantly improved, and the technique has the characteristics of being flexible, safe and efficient.

The invention relates to an acrossocheilus fasciatus gynogenesis device. The acrossocheilus fasciatus gynogenesis device comprises an acrossocheilus fasciatus gynogenesis device body, a base plate, abox, a controller, a control panel, a power switch button, a work indicating lamp, an ultraviolet adjusting knob, a stirrer adjusting knob, a power socket, a partition plate, a high-grade stainless steel disc container, a scale ruler, hollow grooves, steel stirrers, sliding rods, support rods, circular snap grooves, stirring motors, steel stirring rods, stirring blades, a rotating shaft, a flip cover, a flip cover handle and ultraviolet lamps. The acrossocheilus fasciatus gynogenesis device has the advantages that the next generation of parent acrossocheilus fasciatus forms all-female fries byan all-feminization technology; the highest-end scientific technology solves the bottleneck in the aquaculture process of the acrossocheilus fasciatus; meanwhile, second-time meiotic oogoniums at thefinal stage are not released out of polar bodies; and finally, all-female diploid zygotic nucleus eggs are obtained. The effect of the acrossocheilus fasciatus gynogenesis device is designed in a same device, and the whole process of all feminization of the acrossocheilus fasciatus can be carried out. The acrossocheilus fasciatus gynogenesis device is easy to operate, and the feminization rate ofthe acrossocheilus fasciatus is improved.

The invention relates to an induction method for a 'Haida 2#' new variety tetraploid of crassostrea gigas. The induction method comprises the steps of selecting a 'Haida 2#' new variety of crassostreagigas as parent oysters, carrying out manual maturity-promoting culture until the parent oysters are mature, selecting female and male individuals with good gonad development as parent oysters, dissecting for fetching ova, filtering, putting the ova into filtered seawater for maturation, enabling the maturated ova to be fertilized with sperms in the filtered seawater, processing fertilized ova byvirtue of cytochalasin B so as to inhibit the discharging of first polar bodies, after the processing, collecting the fertilized ova, transferring the fertilized ova into an ethanol solution for soaking, finally transferring the fertilized ova into a culture container, and carrying out incubation and larva culture. According to the induction method, the discharging of the first polar bodies of the fertilized ova of the ''Haida 2#'' new variety of a diploid is inhibited by virtue of the cytochalasin B, and the living 'Haida 2#' new variety tetraploids of crassostrea gigas are directly induced,so that the survival rate is greatly increased, and a seedling culturing foundation is provided for the amplified breeding of the 'Haida 2#' new variety tetraploid of crassostrea gigas.

The invention relates to a cell nucleus operation method based on the dynamic drift modeling of the cell nucleus position, comprising the following steps: off-line calibration of the three-dimensional distribution of the cell nucleus relative to a polar body; obtaining the dynamic drift track of the cell nucleus relative to the position of a microneedle through three-dimensional finite element modeling; The factorial design method is used to determine the dominant influencing factors of the dynamic drift of the nucleus position; the relationship between the intracellular movement parameters of the microneedle and the dynamic drift trajectory parameters of the nucleus position is fitted to determine the trajectory of the microneedle required to approach the nucleus, and the microneedle is controlled by the controller. The needle moves in the cell along the determined trajectory and approaches the nucleus and completes the corresponding operations. Using the method for operating the nucleus of the present invention, the enucleation operation can automatically control the trajectory of intracellular movement and the removal amount of cytoplasm. Compared with manual enucleation, the amount of cytoplasm removed is reduced by 60% under the same enucleation success rate, and the cleavage rate of pig cloned embryos is nearly doubled.

The invention relates to a cell nucleus operation method based on cell nucleus position dynamic drifting modeling. The cell nucleus operation method comprises the following steps: calibrating three-dimensional distribution of a cell nucleus relative to a polar body in an off-line manner; obtaining a dynamic drifting track of the cell nucleus relative to the position of a microneedle through three-dimensional finite element modeling; determining dominant influence factors for cell nucleus position dynamic drifting by using a factional factorial design method; and fitting the relationship between microneedle intracellular movement parameters and cell nucleus position dynamic drifting track parameters, determining the microneedle track required for approaching the cell nucleus, and controlling the microneedle to move in the cell along the determined track and approach the cell nucleus by the controller so as to complete the corresponding operation. By utilizing the cell nucleus operationmethod provided by the invention to perform the nucleus removal operation, the intracellular motion track and the cytoplasm removal quantity can be automatically controlled. Compared with a manual nucleus removal operation, the cell nucleus operation method disclosed by the invention has the advantages that the cytoplasm removal quantity is reduced by 60% under the same nucleus removal success rate, and the cleavage rate of cloned domestic pig embryos is nearly doubled.

The invention discloses an oocyte in-vitro maturation culture solution additive and application thereof, and belongs to the technical field of biology. Lactoferrin is found to be capable of remarkably improving the first polar body discharge rate of oocyte, remarkably reducing the ROS content of the oocyte and remarkably improving the blastocyst rate of an embryo after in-vitro fertilization, so that the maturation rate and quality of the oocyte can be remarkably improved, and the lactoferrin can be used for preparing the oocyte in-vitro maturation culture solution additive or an oocyte in-vitro maturation culture solution. The additive and the application can be applied to the embryo engineering technology, and the success rate and quality of embryo engineering are improved.

The invention discloses a method for producing allopolyploid misgurnus anguillicaudatus through inhibiting fertilized egg polar body release. According to the method, diploid misgurnus anguillicaudatus, tetraploid misgurnus anguillicaudatus and paramisgurnus dabryanus are adopted as propagation parents and are used for hybrid fertilized eggs, such that the hybrid fertilized eggs are obtained; on the 5th minute after the hybrid fertilized eggs are obtained, the hybrid fertilized eggs are soaked in ice water with a temperature of 2-3 DEG C for cold shock treatment with a duration of 30-35min; and the eggs are hatched and cultured under room temperature, such that misgurnus anguillicaudatus allopolyploids are obtained. With the method, allotriploid misgurnus anguillicaudatus, allotetraploid misgurnus anguillicaudatus and allopentaploid misgurnus anguillicaudatus can be obtained. With the method, the induced misgurnus anguillicaudatus allopolyploid variety number is high, and the amount is high. The induction success rate is high, and the offspring growth is fast. With the method, the weight of 12-month-age misgurnus anguillicaudatus is increased by an average of -19.99% to 142.34% than that of a normal selfing control group. The weight gain effect is significant. The method has high popularization benefit.

The invention relates to a quantitative enucleation method based on cell orientation, comprising the following steps: S1, modeling the finite element method of the cell nucleus extraction process to determine the distribution of cytoplasm removal required for enucleation under the relative position of the nucleus-microneedle; S2 , stain the cell nucleus and locate the three-dimensional distribution range of the cell nucleus relative to the polar body under the fluorescent field; S3, combine the results of the above two steps to determine the point of a polar body on the focal plane as the cell orientation during the enucleation operation; S4, rotate the cell Draw and quantify the cytoplasm in this direction for enucleation. The present invention utilizes the finite element modeling of the cell nucleus extraction process and the off-line determined cell nucleus relative to the three-dimensional distribution range of the polar body to determine the polar body point during enucleation, and automatically controls the amount of cytoplasm removal, which can achieve 96% of the enucleation amount under 10%. enucleation success rate.

The invention discloses a method for inducing ocean seashell polyploidy by using sodium chloride. The method is characterized by comprising the following steps: selecting the seashell to grow the shell to respectively obtain sperms and ovum; carrying out artificial insemination; continuously observing the developing condition of zygotes through microscopic examination to determine the number of the water body for culturing the zygotes; adding sodium chloride into the culture sea water containing the zygotes according to a proportion that 20-40 go of sodium chloride is added in 1000ml of culture seat water when 40-50 percent of zygotes are observed to release a first polar body or a first zygote is observed to occur the first polar body, so as to prevent the release of a second polar body or the first polar body of the zygotes; evenly stirring and treating for 10-15 min; and transferring the zygotes to normal sea water to incubate . The ocean seashell comprises Pacific oyster, chlamys farreri, Argopecten irradians, Crassostrea rivularis, clam, Scapharca broughtonii. The seashell zygotes are treated by using the sodium chloride and the initiating agent so as to inhibit the release of the polar body and produce the shell polyploidy with no toxicity, efficiency, and simple operation.

The invention provides a method for improving the in-vitro utilization rate of sheep oocyte, which comprises the following steps of: washing poor quality BCB-oocyte obtained by chromosome segregation for 2 to 3 times by using mature culture solution which contains 50 to 100mu Mol / L PDE3 inhibitor milrinone and culturing for 6 to 9 hours, and washing the BCB-oocyte for 2 to 3 times by using matureculture solution which does not contain the milrinone and culturing for 24 hours; removing granular cells around the oocyte, transferring the oocyte to cleaning solution, picking the oocyte and removing a first polar body, and counting the removal rate of the polar body, namely maturation rate; and activating the oocyte from which the first polar body is removed for 4 to 5 minutes by using 5mu Mol / L ionomycin and washing the oocyte for 1 to 3 times by using the cleaning solution, activating the oocyte for 2 to 3 hours by using 2mMol / L 6-methyl aminopurine which is balanced in a CO2 box 2 to 3 hours ago, washing the oocyte for 3 or 4 times by using the culture solution which is balanced in the CO2 box 2 hours ago, culturing the oocyte in the culture solution for 48 hours, counting cleavage rate, and counting blastocyst rate after 168 hours. In addition, the BCB-oocyte obtained by the chromosome segregation is washed by the mature culture solution for 2 to 3 times and cultured for 24 hours to be directly utilized. The method has great practical value for the in-vitro utilization of the sheep oocyte.

The invention discloses a method for chemically inducing an abalone allotriploid and relates to a method for cultivating shellfish. The method comprises the steps: (1) semen abalone selection and maturation promotion; (2) artificial spawning induction; (3) gamete selection and processing; (4) synchronous fertilization; (5) drug induction; (6) drug removal; (7) hatching. Synchronous fertilization is ensured: gametes are strictly prevented from being in contact in advance in the discharging process, it is confirmed through microscopic examination that accidental fertilization does not occur, and the consistency of synchronous fertilization and fertilized egg development is guaranteed. Induction conditions are optimized: 70%-80% of the PB1 proportion of fertilized eggs in a control group is determined as a processing time node by constant microscopic observation, it is ensured that the first polar body is effectively released, and a tetraploid and an aneuploidy are avoided. The stable and efficient induction rate is that the triploid induction rate is 100% or close to 100%.

The invention discloses a method for removing the nucleus of an oocyte through chemical induction. The in-vitro oocyte is induced and treated by decarboxylated colchicine and cycloheximide, and in thefirst division later period of the oocyte after a certain period of time, chromosomes cannot be separated and discharged along with a polar body. The method for removing the nucleus of the oocyte through chemical induction is easy and convenient to operate, a large number of nucleus-free oocytes can be obtained, the nucleus removing efficiency is high, and the method is suitable for application and popularization.

The invention relates to a production method of triploid shellfishes, in particular to a production method of the triploid shellfishes from diploid shellfish production to tetraploid mother shellfish production to triploid shellfish production. The production method is characterized by comprising the following steps of S1, a first stage of fertilization of diploid female eggs and diploid male sperms; S2, a second stage in which the release of a first polar body or a second polar body of fertilized eggs is prevented in the first stage S1 and triploid females are produced; S3, a third stage in which the female eggs in the second stage S2 are fertilized with the diploid male sperms; S4, a fourth stage in which the release of the first polar body is prevented from occurring from the eggs of the third stage S3 and tetraploid males are produced; and S5, the fifth stage in which the diploid female eggs are fertilized with the male sperms in the fourth stage S4, and the triploid shellfishes are generated.

The invention discloses a fixed-point coring method based on augmented reality. The method is characterized in that the position model of the cell nucleus relative to the polar body is obtained by calculating the relative positional relationship between the polar body and the nucleus of the porcine oocyte just matured in vitro under a fluorescent microscope. Use augmented reality technology to mark the position of the nucleus on the oocyte to assist in the enucleation operation. During blind aspiration enucleation, adjust the position of the injection needle by marking the position of the nucleus. It solves the problem that the position of the cell nucleus is not visible during the blind aspiration enucleation process, and realizes the "visualization" effect of the cell nucleus. The success rate of enucleation and the consistency and repeatability of experimental operations are improved.