17 results about "Sperm cryopreservation" patented technology

The invention belongs to the technical field of biology, and particularly relates to an acrossocheilus fasciatus sperm cryopreservation method which comprises the following steps: constructing a simulated water solution for acrossocheilus fasciatus in-vitro fertilization, collecting fresh sperms, detecting the survival rate, preparing a mixed solution of the sperms and anti-freezing components, sub-packaging, cooling and freezing and the like. According to the method, simulated water quality for in-vitro fertilization of acrossocheilus longifin is constructed, the survival rate of sperms observed and detected in the environment is closer to the fertilization condition during thawing activation in the future, and then the sperms with the higher fertilization success rate after thawing are screened out; meanwhile, the constructed simulated water solution can be used as a part of a cryopreservation solution, so that the remaining anti-freezing components can be directly added into the screened sperms, a mixed solution of the sperms and the anti-freezing components can be rapidly prepared under the condition that the service life of the sperms is not remarkably shortened, and the mixed solution is subpackaged, cooled and frozen; the improved anti-freezing component contains trehalose, soybean phospholipid and tea polyphenol, so that the survival rate and activity of the acrossocheilus fasciatus sperms after cryopreservation can be effectively improved.

This utility model relates to the field of cryopreservation equipment technology and discloses a portable mouse sperm cryopreservation device, including a liquid nitrogen tank. A positioning component is fixedly connected to the inner wall of the liquid nitrogen tank, and a measuring rod is slidably connected inside the positioning component. A first foam float is fixedly connected to the bottom of the measuring rod. A hanging groove is provided on the top of the liquid nitrogen tank, and a hanging rod is hung inside the hanging groove. A telescopic rod is slidably connected inside the hanging rod, and a second foam float is fixedly connected to the bottom of the telescopic rod. A placement tray is fixedly connected to the outside of the second foam float, and a pressure cap is slidably connected to the outside of the telescopic rod. This portable mouse sperm cryopreservation device allows for a direct view of the liquid nitrogen level in the liquid nitrogen tank through the cooperation of the measuring rod and the first foam float. The placement tray is connected to the hanging rod via the telescopic rod and is equipped with a pressure cap and limiting components to securely fix the cryopreservation tube.

The invention discloses a cryopreservation method of haliotis diversicolor sperms. The method comprises the following steps: collecting the sperms, testing the initial activity of the sperms, preparing sperm freezing liquid, balancing and freezing the sperm freezing liquid, unfreezing the frozen sperms, and testing the activity after unfreezing. The collected male haliotis discus hannai sperms with the initial activity higher than 90% and a cryoprotectant are mixed according to the volume ratio of 1: 3, and sperm refrigerating fluid is obtained; transferring the sperm freezing liquid into straw, balancing, freezing, and soaking in liquid nitrogen for preservation to obtain frozen sperms; putting the straw filled with the frozen sperms into a water bath kettle, and unfreezing to obtain unfrozen sperms; finally, the straw containing the thawed sperms is cut open, the thawed sperms are transferred to a centrifugal tube, activity testing is carried out, and the activity and quality of the haliotis discus hannai sperms after cryopreservation are remarkably improved through the method.

The invention belongs to the technical field of animal breeding, and particularly relates to application of alpha-ketoglutaric acid or salt thereof in preparation of a sheep semen cryopreservation diluent. The invention discloses the application of alpha-ketoglutaric acid or salt thereof in the cryopreservation of the sheep semen for the first time, and the alpha-ketoglutaric acid or salt thereof can improve the semen quality of the frozen and unfrozen sheep semen and improve the fertilization ability of the frozen semen; such as sperm motility, sperm movement performance, sperm viability, sperm plasma membrane integrity rate, sperm acrosome integrity rate, sperm antioxidant capacity, sperm ATP level, sperm early apoptosis incidence rate, sperm in-vitro fertilized ovulation rate and / or artificial fertilization pregnancy rate, can be used for preparing the sheep sperm cryopreservation diluent.

The invention discloses a method for cryopreserving sperms of oplegnathus punctatus, which comprises the following steps: on the day before the sperms of the oplegnathus punctatus, carrying out drug ripening on male oplegnathus punctatus, on the second day after ripening, collecting seminal fluid by adopting an extrusion method, and collecting the mature seminal fluid into a clean glassware for later use; mPRS diluent is prepared, the MPRS diluent and ethylene glycol are compounded to obtain the anti-freezing liquid, and the mass concentration of the ethylene glycol in the system is 15%; diluting the anti-freezing liquid and the sperm according to a volume ratio of 1: 1 to obtain a sperm mixed liquid; the sperm mixed liquid is balanced for 10 min at the temperature of 4 DEG C, then precooled for 5 min at the position 10 cm away from the liquid nitrogen level, precooled for 5 min above the liquid nitrogen level and finally put into liquid nitrogen for cryopreservation; if the cryopreserved sperm mixed liquid needs to be used, resuscitating and activating the cryopreserved sperm mixed liquid to obtain activated sperms; the survival rate of the sperms after unfreezing and resuscitation is high, and sustainable utilization and development of germplasm resources of the sphagnaceae are better facilitated.

This utility model discloses a sperm cryopreservation tube for sealing, relating to the field of cryopreservation technology. The technical solution includes a tube body with symmetrically fixed blocks on its inner side. A cap is movably connected inside the tube body, and an inner sleeve is fixedly connected inside the cap. A pressing assembly is provided inside the inner sleeve, including a pressing pestle and a button. A sealing sheet is movably connected inside the cap, and a movable plate is fixedly connected to the outside of the sealing sheet. A second spring is fixedly connected to the outside of the movable plate, with one end of the second spring away from the movable plate fixedly connected to the inner wall of the cap. A groove is provided on the inner wall of the tube. By using the pressing assembly, the sealing sheet is pressed, causing it to slide on the inner sleeve and engage in the groove to seal the injection port of the tube body, preventing the cap from falling off and causing liquid nitrogen contamination. This simplifies the process of hand-held rotation when sealing the cap and tube body, quickly sealing the injection port of the tube body, preventing excessive air from entering the tube, and ensuring the sperm preservation effect.

The invention provides ultralow-temperature cryopreservation liquid for white inbred line miniature pig sperms and application of the ultralow-temperature cryopreservation liquid, and belongs to the technical field of pig sperm cryopreservation. The problems that an existing commercial pig sperm cryopreservation technology system and an inbred line white miniature pig cryopreservation technology system are not matched, the sperm quality is reduced and the like are solved. The cryopreservation solution comprises the cryopreservation solution for the sperms of the inbred line white miniature pigs, and the cryopreservation solution is prepared from the following components: a conventional cryopreservation solution, betaine and resveratrol. The method is mainly used for cryopreservation of inbred white miniature pigs.

The invention relates to the technical field of sperm cryopreservation, in particular to a trace sperm cryopreservation method which comprises the following steps: S1, semen collection and pretreatment: collecting a semen sample, and liquefying in a thermostat at 37 DEG C for 30 minutes; detecting the sperm concentration (greater than or equal to 15 * 10 < 6 > / ml), the forward motion rate (PR is greater than or equal to 32%) and the normal form rate (greater than or equal to 4%) by adopting computer-aided semen analysis (CASA); s2, gradient addition of a protective agent: adding a precooled TEST-Yolk buffer solution (containing 10% of glycerol and 0.5 M of sucrose) in three times at the interval of 5 minutes until the final volume ratio is 1: 1; alpha-lipoic acid with the final concentration of 5 mu g / ml is added to inhibit oxidative damage; by avoiding direct contact for gradient cooling, accurate sample positioning and safety protection, the three major pain points of temperature fluctuation, operation risk and sample survival rate reduction in traditional freezing are solved.

The invention belongs to the technical field of cryopreservation, and discloses an optimized diluent composite additive and an application method thereof in sex control frozen sperm cryopreservation, according to the invention, through an orthogonal test, quercetin and vitamin E antioxidants with different proportioning concentrations are compared, and the quercetin and vitamin E antioxidants are grouped as follows: I (3.02 mu g / mL quercetin and 0.1 mg / mL vitamin E), II (15.1 mu g / mL quercetin and 0.1 mg / mL vitamin E), and III (15.1 mu g / mL quercetin and 0.1 mg / mL vitamin E). The antioxidant is added into the control group, III is (3.02 mu g / mL of quercetin and 0.5 mg / mL of vitamin E), IV is (15.1 mu g / mL of quercetin and 0.5 mg / mL of vitamin E), and 0 is (no antioxidant is added, and a blank control group). The dye JC1 in the experiment is an ideal fluorescent probe which is widely used for detecting the mitochondrial membrane potential and can be used for detecting the mitochondrial membrane potential of cells, tissues or purified. When the mitochondrial membrane potential is relatively high, JC1 is gathered in a matrix of mitochondria to form a polymer, and red fluorescence can be generated; when the mitochondrial membrane potential is low, JC1 cannot be gathered in a matrix of mitochondria, and at the moment, JC1 is a monomer and can generate green fluorescence.

The application provides a low-temperature preservation diluent for sperm of Epinephelus lanceolatus and a use method and application thereof. The diluent is prepared from the following raw materials: glucose, calcium chloride, potassium chloride, sodium chloride, sodium bicarbonate, reduced glutathione, sodium dihydrogen phosphate and fetal bovine serum. The diluent contains the following mass of raw materials per liter: 0.4-0.5 g of glucose, 0.03-0.08 g of calcium chloride, 0.15-0.25 g of potassium chloride, 3-4 g of sodium chloride, 1.5-2.0 g of sodium bicarbonate, 1.5-2.5 g of reduced glutathione, 0.008-0.012 g of sodium dihydrogen phosphate, and 8-12% of the fetal bovine serum in terms of the volume percentage of the diluent. The prepared diluent can better maintain the in-vitro activity of sperm, prolong the sperm preservation time, meet the low-temperature preservation requirements of the sperm of Epinephelus lanceolatus, and especially meet the low-temperature preservation requirements of the sperm of Epinephelus lanceolatus.

The invention belongs to the technical field of alpaca artificial insemination, and particularly relates to a cryopreservation agent for alpaca semen, a preparation method and application. The invention provides a cryopreservation agent for alpaca semen. The cryopreservation agent comprises trihydroxymethyl aminomethane, citric acid, fructose, penicillin sodium, streptomycin sulfate, yolk, glycerin and glucose. Wherein the yolk and the glycerol have a synergistic effect, so that the freezing resuscitation rate of sperms is increased; and citric acid and trihydroxymethyl aminomethane are used as buffering agents to stabilize the pH value of the semen diluent, maintain the activity and function of the semen diluent and reduce the damage of free radicals to sperms. The penicillin sodium and the streptomycin sulfate are used for sterilization. The fructose, the yolk, the glycerol and the glucose have an anti-oxidation effect, the cell structure of sperms is stabilized, the activity of the sperms is maintained, and the citric acid and the saccharides are matched to regulate osmotic pressure so as to protect the sperms from being damaged by freezing. The preparation method of the cryopreservation agent is simple and easy, and the sperm motility of the alpaca semen after cryopreservation can be improved.

The invention belongs to the technical field of biology, and particularly relates to a method for improving the cryopreservation effect of donkey sperms, which comprises the following steps: selecting a healthy breeding male donkey with vigorous sexual desire, and collecting the sperm by using a strictly disinfected false vagina; seminal fluid quality detection: immediately evaluating sperm motility of the collected seminal fluid under a 37 DEG C constant-temperature phase contrast microscope; measuring the sperm density by using a sperm density instrument; diluting the semen, and freezing the semen by adopting a programmed cooling method; unfreezing the seminal fluid, taking out the thin tube from the liquid nitrogen, immediately putting the thin tube into a constant-temperature water bath at 38 DEG C, slightly shaking, taking out the thin tube after the seminal fluid in the thin tube is completely unfrozen, wiping water drops on the surface of the thin tube by using sterile gauze, and transferring the seminal fluid into a sterile test tube for later use. According to the invention, sperm motility and fertilization capability are improved: sperm collection and treatment processes are optimized, a proper freezing diluent formula is screened, and freezing is carried out by adopting a programmed cooling method, so that damage of sperms in freezing and unfreezing processes is effectively reduced, and sperm motility and fertilization capability after cryopreservation are remarkably improved.

The invention provides a yak sperm cryopreservation liquid and a preparation method thereof, modified cholesterol is added into the cryopreservation liquid, and the modified cholesterol is obtained by grafting alginic acid with cholesterol and then grafting glutathione. After the modified cholesterol is added into the preserving fluid, a three-dimensional cross-linked network is formed by means of glycerol glycidyl ether, and the network can reduce the size of ice crystals in the freezing process and form a barrier layer outside a sperm cell membrane, so that the ice crystals are prevented from directly damaging the cell membrane and acrosome. Meanwhile, the compatibility of the cholesterol and the sperm cell membrane is excellent, and the modified cholesterol can be enriched on the surface of the cell membrane, so that the effects of filling the membrane gap, enhancing the mechanical strength of the cell membrane and reducing the generation of ice crystals are achieved; meanwhile, glutathione can remove active oxygen, and cell membrane structures and sperm internal components are prevented from being damaged by oxidative stress; in addition, the modified cholesterol can also have a synergistic effect with egg yolk lipoprotein and bovine serum albumin, so that the vitality of frozen sperms is improved, the acrosome integrity rate is improved, and the malformation rate is reduced.

The present invention discloses a method for assessing the activity of Takifugu sperm cryopreservation solution based on computer vision. [Solution] Takifugu sperm are cryopreserved for long periods using a cryopreservation solution, thawed, and then activated. After observation under a microscope, sperm motility videos are recorded. Data augmentation is performed on the video using a binarization method. Based on two deep learning algorithms for target detection and target tracking, the positional changes of multiple Takifugu sperm targets between consecutive frames are tracked and captured, and the significant movements and trajectories of the recognized sperm are analyzed for imitation training. The sperm's movement and viability are determined by statistically analyzing their displacement between different frames. Finally, a visualized sperm viability graph is generated. This method enables long-term cryopreservation of highly active Takifugu sperm, providing technical support for year-round reproduction and species resource conservation. It also improves the accuracy of Takifugu sperm activity detection, simplifies the detection and recognition process, and reduces costs.

Anchovy sperm freezing preservation solution, and a preparation method and application method thereof, belong to the technical field of aquatic germplasm resource protection and breeding, and particularly relate to a method for collecting, preserving and recovering sperm of anchovy fish. The present application is to solve the problems of low efficiency, short preservation time and low sperm activity of the existing anchovy sperm collection, and the anchovy sperm freezing preservation solution is composed of methanol, lecithin, resveratrol and glucose, and is diluted with double distilled water to 1L. The application method is as follows: 1. sperm collection and quality evaluation; 2. preparation of anchovy sperm freezing preservation solution; 3. mixing the sperm with the pre-cooled 4 DEG C anchovy sperm freezing preservation solution, dispensing, and adopting a four-step method for programmed cooling; 4. oocyte collection; 5. sperm warming and activity detection; 6. artificial insemination and hatching. The sperm preservation time is prolonged from several hours to more than several years, and through rapid thawing, the sperm activity can reach more than 40%, and the fertilization rate can reach more than 57%.

The invention discloses a chlamys farreri sperm cryoprotectant and a cryopreservation method, and relates to the technical field of aquaculture. The chlamys farreri sperm cryoprotection liquid comprises dimethyl sulfoxide and 4-hydroxy-2, 2, 6, 6-tetramethylpiperidine-1-oxygen free radicals, a solvent is sterile seawater, the final volume percent concentration of the dimethyl sulfoxide in semen is 4%-8%, and the final concentration of the 4-hydroxy-2, 2, 6, 6-tetramethylpiperidine-1-oxygen free radicals in the semen is 40-60 [mu] M. According to the chlamys farreri sperm cryopreservation liquid and the cryopreservation method, the problems that in the prior art, a chlamys farreri sperm cryopreservation liquid is high in toxicity, complex in procedure and lack of a special scheme are solved, and the chlamys farreri sperm cryopreservation liquid is mainly used for cryopreservation of chlamys farreri sperms so as to achieve long-term preservation of germplasm resources.

The invention discloses application of orientin in improvement of frozen sperm quality and in-vitro cryopreservation of sperms. The invention further discloses a sperm cryopreservation solution, and the sperm cryopreservation solution contains orientin. Orientin is found to have a good protective effect in the sperm freezing process; through multiple mechanisms of reducing oxidative stress, improving mitochondrial functions, improving sperm ultrastructures and the like, the activity and quality of sperms are improved.