32 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 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 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 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 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 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 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 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.