• Title, Summary, Keyword: rapid freezing

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Investigation on the Method of Evaluating the Resistance to Freezing and Thawing of Concrete Subjected Initial Frost Damage (초기동해를 받은 콘크리트의 내동해성 평가법에 대한 검토)

  • 고경택;장일영
    • Journal of the Korea Concrete Institute
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    • v.11 no.4
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    • pp.117-127
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    • 1999
  • In concrete incorporating high volume ground granulated blast-furnace slag that has frozen at early age, to evaluated the results of resistance to freezing and thawing is very difficult because the hydration of the concrete increases over the duration of rapid freezing and thawing test. Hence, the dynamic modulus of elasticity of specimens after freezing and thawing will be favorable results unless the hydration effect is taken into consideration. In this study, a method of evaluating to the resistance to freezing and thawing of concrete subjected freezing at early age, in which the effect of hydration is modified for its increase during rapid freezing and thawing test, is investigated.

Application of Chlorophyll a Fluorescence Imaging Analysis for Selection of Rapid Frozen Sweet Persimmon Fruits (단감(Diospyros kaki)의 동상해 평가를 위한 엽록소 형광 이미지 분석법의 활용)

  • Yoo, Sung Young;Park, So Hyun;Lee, Min Ju;Park, Jong Yong;Kang, Hong Gyu;Kang, Sung Ku;Kim, Tae Wan
    • Korean Journal of Environmental Agriculture
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    • v.34 no.3
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    • pp.210-216
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    • 2015
  • BACKGROUND: In korea, sweet persimmon(Diospyros kaki) cultivation is front to abiotic stresses such as frost damage at fruit maturing stage. The cold and rapid freezing stresses are most damaging to fruit production which is most actively progressed in late fall. This study was performed to evaluate the validity of chlorophyll fluorescence imaging(CFI) technology to determine the degree of frost damage in sweet persimmon fruits. METHODS AND RESULTS: The sweet persimmon fruits were measured separately for each treatment(15, 30, 60 minutes) at 24 hours after treatment(HAT) rapid freezing. A CFI FluorCam (FC 1000-H, PSI, Czech Republic) was used to measure the fluorescence images of the fruits. In rapid freezing for 15 minutes, photochemical parameters were not changed. However, in rapid freezing for 30 and 60 minutes, photochemical parameters were lowered. Especially, $F_m$, $F_v$, $F_v/F_m$ and ${\Phi}PSII$ values were declined under rapid freezing. CONCLUSION: In our study, it was clearly indicated that the rapid freezing could be a stress in sweet persimmon fruits. The CFI analysis and its related parameters are applicable as a rapid assessing technique for the determination of frost damage.

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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COMPARISON OF VIABILITY OF ORAL EPITHELIAL CELLS STORED BY DIFFERENT FREEZING METHODS (구강상피세포의 냉동보관 방법에 따른 세포생존률 비교)

  • Baek, Do-Young;Lee, Seung-Jong;Jung, Han-Sung;Kim, Eui-Seong
    • Restorative Dentistry and Endodontics
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    • v.34 no.6
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    • pp.491-499
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    • 2009
  • This study examined the influence of the storage methods on the viability of oral epithelial cells using conventional cell freezing storage, slow freezing preservation, rapid freezing preservation, and slow freezing preservation with a pressure of 2 Mpa or 3 Mpa. The cell viability was evaluated by cell counting, WST-1 and the clonogenic capacity after 6 days of freezing storage. After 6 days, the frozen cells were thawed rapidly, and the cell counting. WST-1, and clonogenic capacity values were measured and compared. 1. The results from cell counting demonstrated that conventional cryopreservation, slow freezing under a 2 Mpa pressure and slow freezing under a 3 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p < 0.05). 2. The results from the optical density by WST-1 demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05). 3. The clonogenic capacity demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p < 0.05).

Studies on the Survival Rate of Rapidly Frozen Bovine Immature Oocytes (소 미성숙 난포란의 급속동결 융해후 생존성에 관한 연구)

  • 김상근;이봉구
    • Journal of Embryo Transfer
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    • v.10 no.2
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    • pp.115-120
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    • 1995
  • This study was carried out to investigate On in vitro fertilization, survival rate and developmental rate of rapidly frozen bovine immature oocytes. Immature oocytes cultured for 1, 12, 24, 48 hours in 20% FCS + TCM-199 medium and thereafter rapidly freezing-thawed oocytes inseminated with capacitated sperm. The immature oocytes following dehydration by 1.5M DMSO + 2.0M glycerol + 0.25M sucrose + TCM 199 media + 20% FGS were directly plunged into liquid nitrogen and thawes in 3$0^{\circ}C$ water. Rapid freezing embryos co-cultured in 20% FCS + TCM-199 media containing hormones(21U/mL PMSG, 21U /mL hGG and 1 $\mu$g /mL 17$\beta$-estradiol) and cumulus cells(1 x 105-6 cells). Survival rate was defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The in vitro maturation and fertilization rate of immature bovine oocytes on in vitro maturation period(1, 12, 24, 48 hrs) before rapid freezing4hawed were 57.1%, 45.7%, 37.1%, 25.7% and 40.0%, 31.4%, 20.0%, 11.4%, respectively. 2. The survival rate of immature bovine oocytes on in vitro maturation period(1, 12, 24, 48 hrs) before rapid freezing-thawed were 33.3%, 26.7%, 20.0%, and 10.0%, respectively. The survival rate of rapid freezing4hawed immature oocytes was significantly lower than that of non-freezing oocytes. 3. The survival rate of rapid freezing4hawed excellent and good bovine embryos co-cultured in 20% FCS + TCM-199 media containing hormones(PMSG, hCG, 17$\beta$-estradiol) and cumulus cells 4 to 5 hrs and 20 to 24 hrs were 35.0%, 15.0% and 25.0%, 15.0% and 40.0%, 20.0% and 30.0%, 15.0%, respectively. The survival rate of embryos co-cultured in TCM-199 media containing hormones and cumulus cells was significantly higher than that of non co-culture.

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Effects of the Stepwise Exposure Treatments Before Freezing on the Survival Capacity of the Frozen-Thawed Mouse Mature Oocytes by Vitrification or Ultra-Rapid Freezing (동결 전 단계적 노출처리방법이 유리화동결 및 초급속동결-융해 후 생쥐 성숙난자의 생존력에 미치는 영향에 관한 연구)

  • Kim, Sang-Woo;Lee, Jae-Ik;Kim, Mi-Kyung;Lee, Young-Ah;Lee, Kyu-Sup;Yoon, Man-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.2
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    • pp.191-200
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    • 2000
  • Objective: This study was carried out to compare the effects of the stepwise exposure treatments on the morphological normality, fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing and to use as a fundamental data for the cryopreservation of human oocytes. Materials and Methods: The morphological normality and fertilization rates of the vitrified and ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were observed. After choosing the 3step exposure treatment groups, we observed the morphological normality and fertilization, blastocyst formation rate of the vitrified and ultra-rapid frozen mouse mature oocytes. Results: The morphological normality and fertilization rates of the vitrified mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 75%, 85%, 88% and 58%, 61 %, 54% respectively. There were no significant differences among treatments(p>0.05). The morphological normality and fertilization rate of the control was 92% and 65%. There were no significant differences in fertilization rate among control and treatments (p>0.05). The morphological normality and fertilization rates of the ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 83%, 83%, 84% and 75%, 63%, 56% respectively. There were no significant differences among treatments (p>0.05). The morphological normality and fertilization rate of the control was 95% and 67%. There were no significant differences among control and treatments (p>0.05). The morphological normality and fertilization rate of the vitrified or ultra-rapid frozen mouse mature oocytes after 3step exposure treatment were 69% and 75%, respectively. The blastocyst formation rate was 60% and 57%. The results did not differ significantly between vitrification and ultra-rapid freezing (p>0.05). Conclusion: As known in the above results, there were no significant differences in the fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing among the control and treatments. It is suggested that vitrification and ultra-rapid freezing method were effective for the cryopreservation of mouse mature oocytes.

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A Study on the Effects of Cryopreservation by One-Step Straw Method on the Survival of Bovine Embryos (1단계 straw동결법이 소 수정란의 생존성에 미치는 영향에 관한 연구)

  • 김상근;김무강
    • Journal of Embryo Transfer
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    • v.9 no.1
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    • pp.65-71
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    • 1994
  • This study were carried out to investigate the effective concentration of cryoprotectant agents and sucrose by one-step straw method, and to determine the optimum thawing temperature and equilibration time of frozen bovine embryos. The bovine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 3O$^{\circ}C$ water. Survival rate was defined by FDA test. The results are summarized as follows : 1. The survival rate of bovine embryos thawed after rapid freezing in the freezing medium containing a various kinds of cryoprotective agents added 0.25M and O.50M sucrose were 28.6% and 25.0%, 35.1% and 31.6%, 32.4% and 24.4%, 34.2% and 28.2%, 18.9% and 17.6%, 14.7.% and 21.6%, respectively. 2. The survival rate of bovine embryos thawed after rapid freezing in the freezing medium containing a various concentration of sucrose added 1.5M and 2.OM glycerol, i.5M and 2.OM DMSO and 1.5M and 2.OM propanediol were 22.9~37.8%, 2O.7~31. 3%, 19.2~30.0% and 17.2~25.0%, respectively. 3. The temperature thawed at 2$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher embryos survival rate than did at 3$0^{\circ}C$ and 35$^{\circ}C$. 4. The equilibration time on the survival rate of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (1O~20 min.).

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Studies on the Rapid Freezing of Mouse Embryo I. Effects of Cryoprotectants Concentration on the Mouse Embryo Survival of the Rapid Freezing (Mouse 수정란의 급속보존에 관한 연구 제1보 내동제 농도가 Mouse 수정란 급속동결시 생존율에 미치는 영향)

  • 강만종;김영훈;문성호;김종규
    • Korean Journal of Animal Reproduction
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    • v.13 no.3
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    • pp.134-140
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    • 1989
  • Studies were conducted to seek reasonable methods of rapid freezing of mouse embryos using liquid nitrogen. The effects of the cryoprotectants concentration and the substitution of raffinose to sucrose in freezing and dilution medium on mouse embryo survival rates were determined using the FDA-test. The summarized results are as follows : 1. When 0.3M of sucrose was added into the freezing and dilution medium, FDA scores of embryos were 1.48(1.5M), 3.81(3.0M) and 4.10(4.5M). Higher FDA scores of embryos were obtained in 3.0M and 4.5M glycerol concentrations (P<0.05). 2. With the addition of 0.3M raffinose to the freezing and dilution medium, FDA scores of embryos did not significantly differ between glycerol levels ; 3.97(1.5M), 4.11(3.0M) and 3.54(4.5M). Higher scores of embryos existed in 3.0M glycerol concentration. 3. Concentration of sucrose or raffinose in freezing and dilution medium affected FDA scores of embryos. When sucrose concerations of 0.3, 0.5 and 1.0M were added to the freezing medium, FDA scores of embryos were 3.12, 2.38 and 0, respectively. However, when the same concentrations of raffinose were added to freezing medium, the FDA scores were 4.21, 2.91 and 0. In both cases, better FDA scores of embryos were attained in 0.3M of sucrose or raffinose (P<0.01).

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Studies on the Development of Easy Cryopreservation Technique of Bovine Embryos I. Effects of Kinds, Concentration and Freezing Method of Cryoprotectants on In Vitro Developmental Rates of Embryos (소 수정란의 간이 동결기법 개발에 관한 연구 I. 내동제의 종류, 농도 및 동결방법이 체외발생율에 미치는 영향)

  • 김상근;남윤이;현병화;석호봉
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.95-102
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    • 1997
  • The studies on the carried out to investigate the effective concentration of cryoprotectant agents and sucrose by one-step straw method of bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1$\mu\textrm{g}$/ml $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentrations of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival and in vitro developmental rate was defined as devellpmental rate on in vitro culture or FDA-test. The results are smmarized as followes : 1. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was attained 2.0M glycerol, 2.0M DMSO, 1M or 2.0M propanediol. 2. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was obtained single cryoprotectant(6.7~17.4%) than mixed cryoprotectants(6.7~16.7%). 3. In vitro developmental rate of bovine embryos after rapid frozen-thawing in the freezing medium added 0.25M and 0.50M sucrose were higher cleavage rate than those of sucrose concentration of 0.75M and 1.00M. 4. The freezing methods on in vitro developemental rates of bovine embryos was attained slow freezing method(9.70~15.6%) higher than rapid freezing method(9.4~13.3%).

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Effect of Cryoprotectant Concentration and Equilibration Time on Volume Change and In Vitro Development of Intact and Bisected Mouse Embryos following Rapid Freezing (동결보호제의 농도와 평형시간이 생쥐의 정상배 및 분할배의 용적 변화와 체외 발달에 미치는 영향)

  • 이은봉;공일근;강대진;박충생
    • Korean Journal of Animal Reproduction
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    • v.16 no.1
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    • pp.47-53
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    • 1992
  • This study was carried out to investage the effect of cryoprotectant concentration and equilibration time on volume change and in vitro development of intact and bisected mouse embryos by rapid freezing. When compacted morulae were rapidly frozen in 3.0 to 4.0 glycerol or DMSO with 0.25M sucrose solution, the superior(P<0.05) post-thaw survival rate was obtained at the glycerol concentration of 4.0M(89.4%) than 3.0M(71.4%) or 5.0M(42.4%), but at the DMSO concentration of 3.0M(84.5%) than 4.0M(51.1%) or 5.0M(0.0%). The optimal equilibraton time for rapid freezing of ZP-free or bisected morulae in 4.0M glycerol with 0.25M sucrose was found tobe 3 minutes. The minimal volume of compacted morulaewhich corresponded with 61 to 62% of pre-equilibrated embryo volume was obtained from equilibration for 3 minutes in both 3.0 and 4.0M glycerol solutions with 0.25M sucrose.

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