33 results about "Polar body" patented technology

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 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 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 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 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 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.