• Title/Summary/Keyword: Freezing method

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Studies on the Rapid Freezing of Mouse Embryo II. Effects of the Development Stage and Seeding on the Mouse Embryo Survival of Rapid Freezing (Mouse 수정란의 급속동결에 관한 연구 제II보 Mouse수정란 급속동결에 있어서 수정란의 발육단계와 식빙(seeding)이 생존율에 미치는 영향)

  • 강만종;김영훈;문성호;김중규
    • Korean Journal of Animal Reproduction
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    • v.13 no.3
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    • pp.141-148
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    • 1989
  • The effects of seeding method and optimum time for freezing embryos according to the developmental stages on embryo survival rates after rapid freezing were determined using the FDA-test. The summarized results are as follows : 1. In the rapid freezing of embryos, the sucrose added medium together with Co-seeding or non-seeding showed the FDA scores of 4.67 and 4.20, respectively, but, raffinose addition obtained FDA scores of 4.27 and 3.97. 2. The developmental stage of embryos at freezing was most critical on the survival of embryos after thawing. Higher FDA scores were obtained in the order of blastocyst stage(4.94), morula stage(3.82) and ealy stage(2.65) in sucrose added medium. The same trend was observed in the raffinose added medium with an order or 4.91, 4.47 and 2.32. 3. Microscopic study of embryo before freezing and post-thawing indicated that the embryo showed shrinkage within 5 minutes after the embryo was transfer to the freezing medium. When thawed embryo was tranfered to the dilution medium, swelling of the embryo was observed and there after it reshrank indicating the removal of cryoprotectant from the embryo. The size of the embryo recovered to the original state when it was moved into a PBS-solution.

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Current Application of Embryo Cryopreservation for Farm Animals (가축 수정란 동결보존의 최근 이용방법)

  • 석호봉
    • Journal of Embryo Transfer
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    • v.4 no.1
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    • pp.1-13
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    • 1989
  • This paper reviews the most important steps that have generated consistent progress in principles and developmental progress of embryo cryopreservation, and also study on freezing procedure and its application by conventional method and current improved method for freezing procedure and its appilcation of embryo cryopreservation in farm animals. Four were of particular interest: 1.The transport of water across the ccli membrane (zona pellucida) during freezing and thawing accordinglyplays a role in determing whether the celi survives. This movement of water is controlied mainly by extracellular phase changes and by the nature and concentration of any cryoprotective agent present. Therates of cooling, freezing and warming, and the intervals over which they are applied are further decisi've factors in determining whether a cryopreservation procedure allows survival after thawing. 2.The first successful deep freezing experiments with sheep morula and blastocysts during the seventies were based on the early procedures used for mouse embryos.Current research during the eighties is developed with the aim of simplifying and improving current procedures such as one-step dilution and rapid or ultra-rapid cooling by using the model of laboratory animals. 3.The conventional method for the embryo cryopreservation is described. An alternative to this method which may result in high survival and also in reducing of the freezing and thawing time is done by combing a permeable cryoprotectant such as glycerol, DMSO or propanediol and a non-permeable compound such as sucrose, trehalose, raffinose or lactose. 4.Finally a different approach to the preservation of embryos, named vitrification, is introduced. This procedure depends upon the ability of concentrated solutions of cryoprotective agents such as glycerol and propanediol to supercool to very low temperature (-196$^{\circ}C$) during rapid cooling before solidifying without formation of ice. However, more complete data are necessary for successful vitrification of blastocysts.

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Comparison of Vitrification and Slow Freezing for the Cryopreservation of Mouse Pronuclear Stage Embryos (생쥐 전핵기 배아 냉동보존에서 완만동결과 유리화동결의 비교)

  • Kim, Mi-Young;Lee, Yu-Il
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.2
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    • pp.117-124
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    • 2007
  • Objective: The aim of this study was to compare the efficacy of slow freezing with vitrification method for cryopreservation of mouse pronuclear stage embryos. Methods: Mouse pronuclear embryos obtained from superovulated mice and classified into 2 groups of slow freezing and vitrification. Slow freezing solution consisted of 1.5 M PROH, 0.1 M sucrose, while vitrification solution consisted of 40% ethylene glycol, 18% Ficoll and 0.5 M sucrose diluted in Dulbecco's phosphate-buffered saline supplemented with 10% SSS. Recovery and survival rates after thawing and development rates to hatching balstocyst stage were compared between two groups. Results: After freezing and thawing, recovery rate of slow freezing group was 93.8%, whereas vitrification group was 66.5% (p<0.01). Survival rate of recovered embryos were similar between two groups as 83.2% in slow freezing and 87.6% in vitrification. Embryo development rates to 2-cell stage after 24 hrs (77.0% vs 59.1%), 4-cell after 48 hrs (72.6% vs 53.3%), blastocyst after 96 hrs (53.1% vs 40.1%) of thawing were significantly higher in vitrification group than those of slow freezing group, respectively. Conclusion: The vitrification method may provide better developmental competence of frozen-thawed embryos than that of slow freezing method for cryopreservation of mouse pronuclear stage embryos.

An Experimental Study on the Freezing and Thawing of Concrete Mixed with Steel Slag (제강 슬래그를 이용한 콘크리트의 동결융해에 대한 실험적 연구)

  • 정원섭;조성현;권기주;박성우;김진만
    • Proceedings of the Korea Concrete Institute Conference
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    • 2003.11a
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    • pp.48-53
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    • 2003
  • The steel slag, a by-product which is produced on the process of manufacturing steel by refining pig iron is mainly used as road materials after aging it. It is necessary to age steel slag for long time in air because the reaction with water and free-CaO in steel slag could make the volume expanded. Due to this reason it prevents steel slag from being used as aggregate of concrete. But steel slag used in this study is controled by a air-jet method which rapidly cools substance melted at a high temperature. Rapid cooling prevents from generating of free-CaO in steel slag. In this study, it was investigated that steel slag manufactured by air-jet method affects on concrete in the freezing and thawing. As results of this study, concrete mixed with steel slag was worse in the freezing and thawing than concrete mixed with sand in spite of using air entraining agent. To obtain durability of concrete in the freezing and thawing, it is desirable to mix 50% of steel slag in concrete per unit weight of volume.

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Integrity Evaluation of Ice Plugged Pipes Applied on Short Jacket

  • Park, Yeong-Don;Son, Geum-Su
    • Nuclear Engineering and Technology
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    • v.34 no.2
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    • pp.105-116
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    • 2002
  • In special industrial fields such 3s nuclear power plants and chemical plants, it is often necessary to repair system components without plant shutdown or drainage of system having many piping structures which may have hazardous or expensive fluid. A temporary ice plugging method for blocking internal flow is considered as a useful method in that case. According to the pipe freezing guideline of the nuclear power plant, the length of a freezing jacket must be longer than twice of the pipe diameter. However, for applying the ice plugging to short pipes which do not have enough freezing length because of geometrical configuration, it is inevitable to use shorter jacket less than twice of the pipe diameter. In this study, the integrity evaluation for short pipes in the nuclear power plant Is conducted by an experiment and the finite element analysis. From the results, the ice plugging process in short pipes can be safely carried out without any plastic deformation and fracture.

Effects of Sub-freezing Systems on the Freshness of Pork Loin, Beef Loin and Tuna (냉각방식에 따른 반냉동 돼지고기, 쇠고기와 참치의 신선도 변화)

  • Kim, Don;Chang, Young-Ki;Park, Ki-Hwan;Lee, Young-Chun
    • Korean Journal of Food Science and Technology
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    • v.32 no.2
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    • pp.341-348
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    • 2000
  • Effects of different types of sub-freezing methods on quality of raw pork loin, beef loin and tuna were studied. Storage tests were undertaken as follows; the three different types of sub-freezing methods, such as regular cold chamber at $-1^{\circ}C$, air-blast sub-freezing at $-5^{\circ}C$ and sharp-plate sub-freezing at $-5^{\circ}C$, were evaluated for extending freshness of samples. Pork loin packed with polyethylene-film showed no significant differences (P<0.05) in physico-chemical properties among the above sub-freezing methods. Air-blast sub-freezing methods revealed faster evaporation of moisture from samples. The same sub-freezing method also showed increased VBN values of pork loin and tuna samples. Regular cold chamber resulted in increased TBA values and K values of beef loin and tuna, respectively. It appeared that sharp-plate freezing system kept the food samples with the least loss of freshness among the above three sub-freezing methods. These results indicated that freshness of raw pork, beef and tuna could be significantly extended by the sharp-plate sub-freezing method.

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The Hatching Rate of Resting Eggs of the Rotifer Brachionus plicatilis according to Preservation Method (보관 방법에 따른 Rotifer Brachionus plicatilis 내구란의 부화)

  • Youn, Joo-Yeon;Hur, Sung-Bum
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.6
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    • pp.665-670
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    • 2011
  • The rotifer Brachionus plicatilis is one of the most important food organisms in aquaculture. The resting eggs produced by mictic female rotifers are easily stored and hatched, making them useful as the starter for the mass culture of rotifers in marine larval culture. This study examined the optimum preservation method for resting eggs to ensure a high hatching rate. To produce resting eggs, the marine rotifer B. plicatilis was cultured with Nannochloris oculata (KMMCC 16). The resting eggs were harvested and cryopreserved using 5% and 10% methanol (MeOH), dimethylsulfoxide (DMSO), and glycerol as cryoprotectant agents (CPAs). The cryopreservation comprised slow or rapid freezing and the resting eggs were stored for one month in liquid nitrogen ($-196^{\circ}C$). The resting eggs were also dried at different temperatures (30, 40, and $50^{\circ}C$) and for different times (1, 2, and 3 h). In general, the hatching rates of the resting eggs preserved with CPA were higher than those without CPA and the slow freezing method was better than the rapid freezing method. However, the optimum CPA concentration for the hatching rate of the resting eggs varied with the freezing method and kind of CPA, and the CPA also affected the viability of the resting eggs. Dried resting eggs had a high, rapid hatching rate over 80%. The moisture content of the resting eggs cryopreserved in liquid nitrogen affected the hatching rate. Drying at $30^{\circ}C$ for 1 hour resulted in a high hatching rate of the resting eggs. In conclusion, drying at $30^{\circ}C$ for 1 hour and preservation in liquid nitrogen with the slow freezing method, without CPA, is recommended for a high hatching rate (ca. 95%) of rotifer resting eggs.

Studies on cryopreservation of nuclear transplanted mouse embryos (마우스 핵이식란의 동결에 관한 연구)

  • Lee, Byeong-chun;Jo, Choong-ho;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.171-178
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    • 1993
  • The present study was carried out to investigate the developmental potency to blastocyst after freezing and thawing of nuclear transplanted 2-cell embryos. The nuclei from 2-, 4- and 8-cell mouse embryos were transferred into enucleated 2-cell embryos, and the reconstituted embryos were submitted to direct current(DC) pulse at output voltage of 2.0 kV/cm for $100{\mu}$ sec to induce cell fusion. The recovery rate and developmental potency to blastocyst after freezing and thawing of nuclear transplanted 2-cell embryos was investigated. 1. The recovery rate of nuclear transplanted 2-cell embryos in normal morphology after freezing and thawing was significantly higher in rapid freezing(DMSO 4.5M) than in slow cooling(p<0.01). 2. When the recovered embryos in normal morphology were cultured in vitro, there were no significant differences in the developmental potency to blastocyst between the freezing methods and the concentrations of cryoprotectant. In summary, these experiments have proved that rapid freezing method(DMSO 4.5M) is effective in nuclear transplanted 2-cell mouse embryos. If improved micromanipulation techniques and freezing are combined, nuclear transplantation technique will contribute to the improvement of productivity in livestock animals.

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The Freezing Process of the Water Supply Pipe in an Exterior Wall (외벽측 급수관의 동결 과정에 관한 연구)

  • Kang, Han-Gi;Lee, Jae-Heon
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.19 no.11
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    • pp.782-788
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    • 2007
  • In this paper, the freezing process of the water supply pipe in the exterior wall of an apartment house was analyzed by numerical method. The thickness of the pipe insulation and the percentage of insulation damage were considered as parameters in this paper. In the cases of the 0%, 8% and 20% damaged of the 5mm thickness insulation, the freezing was completed after 13 hours, 10 hours and 7 hours respectively. And in cases of the 10mm thickness insulation, the freezing was completed after 18 hours, 10.5 hours and 8 hours respectively. As a result, it is predicted that the water freezing would occurred when the water supply pipe with 8% or 20% damaged insulation are installed in the exterior wall. However, the water freezing would not occurred when the water supply pipe with 10mm thickness insulation of 0% damage is installed in the exterior wall.

Evaluation of freezing and thawing damage of concrete using a nonlinear ultrasonic method

  • Yim, Hong Jae;Park, Sun-Jong;Kim, Jae Hong;Kwak, Hyo-Gyong
    • Smart Structures and Systems
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    • v.17 no.1
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    • pp.45-58
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    • 2016
  • Freezing and thawing cycles induce deterioration and strength degradation of concrete structures. This study presumes that a large quantity of contact-type defects develop due to the freezing and thawing cycles of concrete and evaluates the degree of defects based on a nonlinearity parameter. The nonlinearity parameter was obtained by an impact-modulation technique, one of the nonlinear ultrasonic methods. It is then used as an indicator of the degree of contact-type defects. Five types of damaged samples were fabricated according to different freezing and thawing cycles, and the occurrence of opening or cracks on a micro-scale was visually verified via scanning electron microscopy. Dynamic modulus and wave velocity were also measured for a sensitivity comparison with the obtained nonlinearity parameter. The possibility of evaluating strength degradation was also investigated based on a simple correlation of the experimental results.