• Title/Summary/Keyword: Freezing Solution

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Vitrification and Ultrarapid Freezing of Day 2 Mouse Embryos (제 2일째 생쥐 배아의 초자화동결과 초급속동결)

  • Yang, Jung-Sook;Sohn, Cherl;Bae, In-Ha
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.3
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    • pp.283-289
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    • 2000
  • Objective: The study was performed to compare the survival rate and the development of day 2 mouse embryos which had freezing procedures done. Methods: We used three different vitrification solutions (EFS, VS14, DPS) and a ultrarapid freezing solution (UFS) for cryopreservation of day 2 mouse embryo. Results: We tested toxicity by exposing embryos to vitirification solutions and a ultrarapid freezing solution. The survival rates are 100%, 97.8%, 95.6% and 100% (EFS, VS14, DPS and UFS). After cultured for 96 hours, hatching rates of each group are 93.5% (no freezing), 95.6% (EFS), 86.4% (VS14), 93.0% (DPS), and 93.0% (UFS). There is no significant differences among groups. The survival rates after thawing cryopreserved embryos are 80.2%, 91.7%, 69.5%, 0% and 91.8% (slow freezing, EFS, VS14, DPS and UFS). Also cultured for 96 hours, the hatching rates are 93.5% (no freezing), 84.1% (slow freezing), 93.9% (EFS), 48.5% (VS14) and 70.1% (UFS). Conclusion: The survival rates of vitrification in EFS solution and ultrarapid freezing are higher than slow freezing (p<0.05). The hatching rate of vitrification in EFS solution cultured for 96 hours is highest, so vitrification of day 2 mouse embryos in EFS solution considered as more effective for cryopreservation.

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Effects of Freezing Conditions on the Concentration-Efficiency in the Progressive Freeze-Concentration (Progressive Freezing에 의한 동결 농축법에 있어서의 농축효과에 미치는 동결조건의 영향)

  • 배승권
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.6
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    • pp.984-989
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    • 1995
  • The concentration-efficiencyh of blue dextran solution in the progressive freeze-concentration was related to the freezing conditions such as the freezing speed and the stirring speed in the solution phase. From the theoreticla balance equation of heat and mass transfer at freezing front, the relationship between the freezing conditions and the ice structure at freezing front was drived. A high freeze-concentration efficiency was obtained under the operating conditions represented by a low speed of freezing and a high speed of stirring. The operating conditions were related to a smooth solid-liquid interface and these results were well explained by the theoretical equation. Effect of the solute component size on the concentration efficiency in the progressive freezeconcentration was also tested. The concentration efficiency of latex particles showed a lower value than that of blue dextran, however, its difference was insignificant.

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Establishment of Freezing Conditions for Improving Cryosurvival in Miniature Pig Spermatozoa (미니돼지 정액의 동결-융해 후 생존율 향상을 위한 동결 조건 확립)

  • Lee, Yong-Seung;Yoo, Han-Jun;Cheong, Hee-Tae;Yang, Boo-Keun;Woo, Jea-Seok;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.329-334
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    • 2011
  • This study was conducted to establish a freezing method of miniature pig spermatozoa. The semen 더aculated from PWG M-type miniature pig was collected by gloved-hand method. The semen was diluted with same volume extender (m-Modena B). The frozen solution used frozen solution of four different (LEY, TCG, BF-5 and m-Modena+egg yolk) for find optimal frozen solution in miniature pig sperm. The diluted semen for frozen rate assay was added to LEY solution (solution I: 11% lactose+egg yolk; solution II: solution I+glycerol+OEP), and frozen depending on freezing rate by the three different freezing methods (A: until $5^{\circ}C$ for 1 hrs, holding at $-102^{\circ}C$ for 10 min; B: until $5^{\circ}C$ for 2 hrs, holding at $-102^{\circ}C$ for 10 min; C: until $5^{\circ}C$ for 3 hrs, holding at -80 and $-102^{\circ}C$ for 10 min). Semen cooled until $5^{\circ}C$ was added with glycerol 1, 3 and 5%, and take a equilibrium time for 0, 10 and 30min. Frozen-thawed sperm were evaluated for viability, acrosomal status and morphological abnormality. The results of frozen-thawed sperm ability by frozen solution, viability was higher in LEY solution compared to other three different frozen solution. AR pattern of LEY solution were lower than other three different frozen solution. The results of freezing rate, viability was higher in B method compared to other methods (p<0.05). Acrosomal statute was intacted in A and B methods than C method. The experiment for glycerol condition was showed that sperm viability was higher in extender with 1% and 3% glycerol and equilibrium time of 0 min. The acrosome damage was lower in extender with 1% glycerol and equilibrium time of 10 min than other conditions. In conclusion, the optimal conditions for cryopreservation of miniature pig spermatozoa obtained in LEY frozen solution, cooling rate of 1~2 hrs, 1~3% glycerol concentrations and glycerol equilibrium time of 0~10 min.

Studies on the soil freezing depth and change of moisture contents in evergreen plants upon subzero temperature in (강원도지역의 토양동결심 및 상록식물의 함수량 추이에 관한연구 (1))

  • 홍종운;허범양;원경열;임병춘;이기철;하상건
    • Asian Journal of Turfgrass Science
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    • v.4 no.1
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    • pp.42-48
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    • 1990
  • Experiments were conducted to investigate the soil freezing depth and pattern with freezing measuring instruments during 1988-l989 winter season in Kangwon province. Freezing measuring instrument was made with acrylic pipes which were consisted of inner and outer parts. Inner pipe was filled with 0.01 % methylene blue solution and rubber hose to protect pipe breakdown by solution freezing. Freezing measurements were carried out by observing discoloration of methylene blue solution. Moisture content of evergreen trees and ground cover plants was also examined in the winter season. The observed results are as follows: 1.In the land of I OOM above sea level, soil freezing depth became deeper as the sum of Accumulated degree-days of temperature below 0˚C(0˚C . day) increased: Soil freezing depth was 30-40cm at l00˚C, 42-43cm at 150˚C, and 47cm at 200˚C day 2.Soil freezing with vinyl mulching was less developed by l3cm at l00˚C with sum of subzero temperature, by l7cm at 200˚C than that of the bare ground. Soil of rich hulls mulching with 4Ocm was not frozen until soil freezing at the bare ground was developed to 25cm depth. 3.Cashmeron mulching was more effective than felt mulching in the heat insulation of soil. 4.Thawing of soil was done from the lowest part of the frozen in the ground to upward in the beginning and after that it was done from the surface of frozen soil to downward. Finally thawing was completed at the middle of frozen soil.

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Experimental Study on the freezing of Aqueous Binary Solution Saturated Packed bed in a Square Cavity (다공성 물질이 충전된 정방형 밀폐용기 내에서 수용성 혼합용액의 동결거동에 관한 실험적 연구)

  • Choi, J.Y.;Kim, B.C.
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.4 no.3
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    • pp.175-182
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    • 1992
  • Freezing of the binary solution ($H_2O-NaCl$) saturating a packed bed of spheres is investigated experimentally. The system is cooled through its top surface, and the bottom is maintained at a temperature above the liquidus. Experiments are performed on the hypolutectic side, and the cold wall temperature is lower than the eutectic point. The effects of initial mixture concentration, superheat and glass bead diameter on temperature and concentration distributions are investigated. Supercooling was observed only at early times of the freezing process for experiments with 5% initial salt concentration. Flow visualization experiments and mushy-liquid interface position observations revealed natural convection in the liquid region. Remelting phenomena was not observed at both the solid-mushy and mushy-liquid interfaces.

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Freezing and Melting Phenomena of Urea-water Solution for Diesel Vehicle SCR System (디젤차량 SCR 시스템용 요소수용액의 동결과 해동 현상)

  • Choi, B.C.;Seo, C.K.;Myong, K.J.
    • Journal of Power System Engineering
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    • v.13 no.4
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    • pp.5-10
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    • 2009
  • Urea-SCR system, the selective catalytic reduction using urea as reducing agent, is a powerful technique to reduce nitrogen oxides(NOx) emitted from diesel engines. However, a tank of urea(32.5 wt%)-water solution can be frozen in low ambient temperature levels of below $-11^{\circ}C$. The purpose of this study is to understand freezing and melting phenomena of the urea-water solution, and its can be applied to get the urea-water solution from frozen it within 5 minutes after cold start. Factors considered were the type of heater and the urea tank shape. From the results, it was found that melting volume of cartridge heater B during 5 minutes of heating period was 83ml when supplying electric power of 150W. Horizontal heater B, which was put in the narrow bottom space of the tank T1, had fast melting characteristics.

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A Fundamental Study on the Effect of Anti-Freezing Agent of Concrete (콘크리트용 내한제의 효과에 관한 기초적 연구)

  • 윤기원;조병영;한천구;김무한
    • Proceedings of the Korea Concrete Institute Conference
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    • 1994.04a
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    • pp.163-168
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    • 1994
  • If the concrete is exposed below freezing point of the outer temperature before hardened, the quality of the concrete will be lowered after hardened. Anti-freezing agent which doesn't corrode the steel bar and doesn't generate the alkali-aggregate reaction by noncholride recently developed in Japan and Northern Europe. But the effects of these agents are rarely known. Therefore, this study is desigend for analyzing the freezing properties at the condition of solution of solution and cement paste. And this study aims to present the reference data for practical use of the concrete works using anti-freezing agent.

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Effect of Supplementation of Trehalose, Glycerol on Conventional Freezing and Vitrification of Boar Sperm

  • Choi, Sun-Ho;Lee, Mi-Jin;Lee, Kyung-Mi;Sa, Soo-Jin;Kim, Hyun-Jong;Jin, Hyun-Ju;Song, Yong-Sup;Park, Jun-Cheol
    • Journal of Embryo Transfer
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    • v.29 no.4
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    • pp.397-401
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    • 2014
  • The boar sperm has more lipid droplets and specialty of seminal plasma compared with other species, causing difficulties of freezing sperm and decreases for the utilization of frozen semen into the artificial insemination. However, several studies reported significant results for the recovery of sperm motility and reproductive by addition of cryoprotectants and seminal plasma after thawing. This study was designed to investigate the effects of supplementation of trehalose or glycerol in the LEY (lactose and egg yolk in BTS) solution for the conventional freezing and vitrification process. Two boars aged 16 months were used to collect semen for 2 times in a week. The samples were allotted to 3 freezing solutions (LEY + glycerol 10.5% + OEP 1.5%, LEY + trehalose 1M + OEP 1.5%, and sucrose 1.5M + trehalose 1 M + OEP 1.5%) after centrifugation at 800 g for 10 minutes. Semen was equilibrated in freezing solutions for 10 minutes and injected into plastic straws with 2~3 air bubbles to minimize freezing damages. Vitrification was performed to locate sperm in 5 cm above $LN_2$ for 5 minutes, and the conventional freezing was conducted with an automatic freezer. Motility and survival rates were measured by CASA (Computer assisted sperm an alyzing system) and FITC (Fluorescein isothiocyanate), respectively after thawing semen at $50^{\circ}C$ for 12 seconds. The results were analyzed by ANOVA with STATVIEW statistical program. The vitrificatioin solution (LEY + 10.5% glycerol + 1.5% OEP) presented higher motility (20.9%) than other solutions while the solution (LEY + 1M trehalose + 1.5% OEP) showed the lowest (motility : 5.2%). However, survival rates of vitrified sperms detected by FITC showed 1~4% live sperms in almost of dead sperms at all vitrification solutions' groups, but survival rate of freezing solution of LEY + 1M trehalose + 1.5% OEP LEY and LEY + 10.5% glycerol + 1.5% OEP were showed 49%, and 79%, respectively. There were differences (P<0.05) survival rate of conventional freezing in LEY + 10.5% glycerol + 1.5% OEP and LEY + 1M trehalose + 1.5% OEP and the remaining showed no differences. The results suggested that vitrified boar semen was not enough to be utilized for the artificial insemination, but it showed possibility to utilize for ICSI and conventional freezing with glycerol would be useful method for artificial insemination in pig while we choose the outstanding semen against tolerance to freezing damages.

Comparison of the Efficiency between Slow Freezing and Vitrification Method for Cryopreservation of Human Embryos (인간 수정란의 완만 동결과 유리화 동결의 비교)

  • Kim, Eun-Kuk;Kim, Mi-Yeon;Son, Sun-Mi;Kim, Dong-Won
    • Journal of Embryo Transfer
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    • v.23 no.1
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    • pp.19-24
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    • 2008
  • The purpose of this study was to compare the efficiency of slow freezing with that of vitrification method for the cryopreservation of human embryos. Human embryos were derived from in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and the mixed solution of propanedial (1.5, 1.0, 0.5M PROH) and sucrose (0.1M), ethylene glycol (7.5, 15%), dimethyl sulfoxide (7.5, 15% DMSO), sucrose (0.5, 1.0M) and SPS (Serum Protein Substitute) was used for a cryoprotectant for slow freezing and vitrification solution, respectively. Rates of recovery after thawing, morphological normality, post-thaw viability, arrest, morphological abnormality and preimplantation development were compared between two protocols. After freezing-thawing, recovery and survial rate of slow freezing was (88.6% and 73.4%), whereas vitrification was (99.2% and 96.2%) (p<0.05). The arrest rate of slow freezing was significantly lower compared with those of vitrification(8.7% vs 29.9%) (p<0.05). Preimplantation development to the 2-cell (83.8% vs 67.7%), 4-cell (69.0% vs 47.2%) and 8-cell (62.4% vs 37.8%) stages 24, 48 and 72 h after thawing, respectively, were higher in the slow freezing than the vitrification. After slow freezing and vitrification of human embryo at 2-8cell stage, the rate of recovery rate, survival rate and partial damage rate were 92.0% vs 100%, 80.4% vs 96.2% and 52.2% vs 19.0%, respectively. And partial damage rate was significantly lower than those of slow freezing method (p<0.05). These results demonstrate that a slow freezing using PROH is more efficient than a vitrification for cryopreserving the human zygotes, although the vitrification yielded better recovery, survival and partial damage of frozen-thawed 2-8 cell stage embryos than slow freezing method.

A Study on the Freezing and Strength Properties of Cement Mortar using Accelerator for Freezing Resistance (내한촉진제를 이용한 시멘트 모르터의 동결 및 강도특성에 관한 연구)

  • 박상준;김동석;원철;이상수;김영진
    • Proceedings of the Korea Concrete Institute Conference
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    • 2000.10b
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    • pp.1267-1272
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    • 2000
  • When fresh concrete is exposed to low temperature, the concrete may suffer frost damage due to freezing at early ages and strength development may be delayed. These are problems on cold weather concrete. One of the solution methods for resolving these problems has been to reduce the freezing temperature of concrete by the use of chemical admixtures called Accelerators for freezing resistance. Therefore, in this study, we executed freezing temperature of mortar, setting and strength properties with using water reducing accelerator and accelerators for freezing resistance which are producted internationally. As a result of this experiment, the freezing temperature of mortar is lower and the setting property is promoted when the admixing content of accelerators for freezing resistance is increased. Moreover, the compressive strength of mortar used accelerators for freezing resistance represented the result which is similar with result of analysis of compressive strength increase with using logistic curve formula, but in the case of plain and using water reducing accelerator, there is no relation between logistic curve formula, maturity and compressive strength.