• Title/Summary/Keyword: Rapidly thaw

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Cryopreservation of winter vegetation buds of Betula platyphylla var. japonica in liquid nitrogen (자작나무 동아의 액체질소 내 초저온 보존)

  • 안영희
    • Korean Journal of Plant Resources
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    • v.15 no.2
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    • pp.89-95
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    • 2002
  • In woody plant germplasms, using prefrozen dormant buds for materials is one way to achieve successful cryopreservation. The protocol of cryopreservation for White birch (Betula platyphylla var. japonica) winter vegetative buds is the following. First, the branches of White birch were collected in January 20, when the vegetative buds were still in a state of quiescence. The winter buds with about 5㎜ of xylem tissue were removed from the branches. They were dehydrated to moisture contents about 44% by air dry treatment. The buds were prefrozen, with the temperature being decreased by 5∼-20$\^{C}$ and then transfered to the LN(liquid nitrogen) maintained below -l96$\^{C}$. After cryopreservation, the vegetative buds were rapidly thawed in a water bath at 40$\pm$5$\^{C}$. In this case, the cell survival rate of samples was about 86%. After sterilization, buds were then cultured on MS medium. These results demonstrate the feasibility for cryopreservation of winter vegetation buds of Betula platyphylla var. japonica.

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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A Case Study on Chloride Corrosion for the End Zone of Concrete Deck Subjected to De-icing Salts Added Calcium Chloride (염화칼슘이 함유된 제설제로 인한 콘크리트 바닥판 단부의 염해에 관한 사례 연구)

  • Chung, Jee-Seung;Kim, Bo-Heon;Kim, Il-Sun
    • Journal of the Korean Society of Safety
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    • v.29 no.6
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    • pp.87-93
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    • 2014
  • In this study, the reinforced concrete rahmen bridge damaged by the chloride attack was investigated. According to the investigation, the degraded concretes on cantilever kerb and end part were intensively observed. Thus, the chloride content test and half-cell method were performed to evaluate the degraded parts. As a result, the contents of chloride on degraded parts were C and D grade. On the other hand, the half-cell potential values of rebar in degraded concrete were measured with the minor corrosion. This rebar corrosion is expected to progressing. Chloride content D grade is due to expansion pressure by corrosion of rebar and freeze-thaw by permeate water, could see progresses rapidly degradation. In order to prevent chloride attack to concrete deck caused by deicing salts, corresponding to the chloride critical concentration must maintain grade b or at least grade c. Chloride condition evaluation standard apply to evaluation of marine structure chloride attack with chloride attack by deicing salts.

Changes in Quality of Hanwoo Bottom Round under Different Freezing and Thawing Conditions (한우육의 냉동 및 해동 조건에 따른 품질 변화)

  • Chun, Ho Hyun;Choi, Eun Ji;Han, Ae Ri;Chung, Young Bae;Kim, Jin Se;Park, Suk Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.2
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    • pp.230-238
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    • 2016
  • This study examined the effects of freezing and thawing conditions on quality of Hanwoo bottom round. The beef samples were frozen by air blast freezing at $-20^{\circ}C$ or ethanol immersion freezing at $-70^{\circ}C$ and then stored at $-20^{\circ}C$ for 10 days. After 10 days of storage, the frozen samples were thawed with air blast thawing at $4^{\circ}C$ or water immersion thawing at $4^{\circ}C$ and subjected to subsequent analyses of drip loss, water holding capacity, thiobarbituric acid reactive substance (TBARS), volatile basic nitrogen (VBN), total aerobic bacteria, and microstructure. Drip loss significantly increased in samples treated with air blast freezing compared to ethanol immersion freezing, whereas freezing and thawing processes had no significant impact on water holding capacity of the samples. Thawing conditions had a much stronger influence on the TBARS and VBN of the samples than freezing conditions. There was no significant difference in the population of total aerobic bacteria among the four samples subjected to one freeze-thaw cycle. In addition, to analyze the effects of freeze-thaw cycle on the quality of beef, three freeze-thaw cycles were performed during storage. Multiple freeze-thaw cycles increased drip loss, TBARS, and VBN and decreased water holding capacity, accelerating microstructural damage. These data indicate that Hanwoo bottom round can be rapidly frozen and thawed by using ethanol immersion freezing and water immersion thawing methods with minimal impact on meat quality.

Effect of Equilibration Time and Cell Stage on the Survival of IVF Bovine Embryos Cryopreserved by Vitrification (한우 체외수정란의 동결보존시 평형시간과 배 발달단계가 생존성에 미치는 영향)

  • 공일근;주영국;이은봉;김용권;박충생
    • Journal of Embryo Transfer
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    • v.9 no.1
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    • pp.7-14
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    • 1994
  • The present experiments on cryopreservation were designed to examine the effects of solution toxicity, equilibration time and cell stages on the post-thaw survival of bovine IVF embryos. The oocytes were matured in vitro(IVM) for 24 hrs. in TCM-199 supplemented with 35 $\mu$g /ml FSH, 10 $\mu$g /ml LH, 1 $\mu$g /ml estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% $CO_2$ in air. They were fertilized in vitro(IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro(IVC) with bovine oviductal epithelial cells for 7 to 9 days. The bovine IVF embryos were exposed to the EFS solution in one step at room temperature, kept in the EFS solution during different period for toxicity test, vitrified in liquid nitrogen, and thawed rapidly. 1. after the bovine blastocysts were exposed to EFS solution for 2 min. at room temperature and then they were washed in 0.5 M sucrose solution and TCM-199, they were cultured to examined cryoprotectant induced injury during exposure, Most of the embryos(95.0%) developed to reexpanded blastocoels. However, when the exposure time was extended to 5 and 10 min, these development rates dropped dramatically in 5 min. (69.5%) and 10 min. (47.4%), respectively, 2. When the bovine IVF embryos were vitrified in EFS solution after the equilibration for 1 and 2 min. exposure, The embryos to have reexpanded blastocoels following thawing, washing and culture processes were found to he 82.6 and 73.9%, respectively. However, when the exposure time was extended to 3 min, this survival rate dropped to 18.2%. The optimal time for equilibration of bovine IVF blastocysts in EFS solution seemed to he 1~2 min. 3. When the bovine IVF embryos were equilibrated for 1 min. the significantly (P<0. 05) higher post-thaw survival rates were obtained from the embryos of blastocyst stage(81.3%) than morulae stage(5. 1%). The optimal cell stage for viterification with EFS solution proven to he blastocyst stage in bovine IVF embryos. 4. The number of blastomeres of blastocyst stage was examined with nuclear staining with Hoechst 33342 during 7 to 9 days post-insemination. The cell counts of frozen bovine IVF embryos were found significantly(P$\geq$7.5 and those of the fresh embryos 76.6$\geq$7. 1, which were cultured in the sarne period and conditions as frozen embryos.

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Variationsin Air and Ground Temperatures During a Frozen Season in the Subalpine Zone of Mt. Halla (한라산 아고산대의 동결기 기온 및 지온변화)

  • Kim, Taeho
    • Journal of The Geomorphological Association of Korea
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    • v.20 no.3
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    • pp.95-107
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    • 2013
  • In order to examine the temperature regime responsible for periglacial processes, air and ground temperatures were monitored from October 2010 to May 2011 at a subalpine bare patch (1,710m asl) of Mt. Halla. Four thermistor sensor probes were installed at 55 cm above a ground surface and depths of 2 cm, 10 cm, and 20 cm, respectively. A mean air temperature is $-0.1^{\circ}C$, while mean ground temperatures are $1.8^{\circ}C$ at 2 cm, $2.6^{\circ}C$ at 10 cm and $3.2^{\circ}C$ at 20 cm deep. A mean monthly ground temperature at 2 cm deep demonstrates below $0^{\circ}C$ successively from January to March, while those at 10 cm and 20 cm deep show no sub-zero temperature. A total of 72 freeze-thaw cycle was observed in air temperature. However, the numbers in ground temperature reduced into 17 at 2 cm, 8 at 10 cm, and 3 at 20 cm deep. The cycles of air temperature and ground temperature at 2 cm deep mostly fluctuated diurnally, while those of ground temperature at 10 cm and 20 cm deep exhibited a several-daily oscillation. Snow cover over 55 cm high remained from January to early April, and it seemed to disappear completely on April 16. A seasonal frost of at least 2 cm thick was formed on late December and the isotherm of $0^{\circ}C$ descended slowly into 10 cm deep on late March to early April due to the insulating snow cover. It showed the maximum freezing depth of 20 cm on April 7 to 14 and then thawed rapidly so that the frozen ground did not longer after April 17. Periglacial processes are predominant during a freezing period than a thawing period when the ground surface is still covered with snow. The periglacial mass movement in the subalpine zone of Mt. Halla is mainly generated by frost creep in terms of the occurrence depth of diurnal freeze-thaw cycle and the maximum freezing depth of ground.

Effect of Equilibration Tine and Developmental Stages on the Survival of Mouse Embryos Cryopreserved by Vitrification in EFS Solution (Ethylene Glycol을 이용한 유리화 동결시 평형시간과 배 발달단계별 생쥐 배의 생존성)

  • 공일근;정기화;노규진;조성근;이은봉;박충생
    • Journal of Embryo Transfer
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    • v.9 no.2
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    • pp.173-180
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    • 1994
  • The present experirnents on cryopreservation were carried out to investigate effect of solution toxicity, equilibration time and cell stages on the post-thaw survival of mouse morulae and blastocyst embryos cryopreserved by vitrification in EFS solution. The mouse embryos were exposed to the EFS solution in one step at room temperature, kept in the EFS solution during different period for toxicity test, vitrified in liquid nitrogen and thawed rapidly. After the mouse morulae embryos were exposed to EFS solution for 2 and 5 ruin. at room temperature and then they were washed in 0.5 M sucrose solution and basal mediurn(D-PBS + 10% FCS), they were cultured to examined cryoprotectant toxicity induced injury during exposure, most of embryos developed to expanded blastocysts(100 and 90.0%). However, when the exposure time was extended to 10 and 20 min, these development rates dropped dramatically in 10 ruin. (75.0%) and 20 ruin. (4.5%), respectively. When the compacted morulae were vitrified in EFS solution after equilibration for 2 and 5 min, the embryos have developed to normal blastocyst following thawing, washing and culture processes was 89.3 and 89.6%. However, when the exposure time was expanded to 10 ruin, this survival rate dropped to 68.8%. When the blastocyst were vitrified in EFS solution after equilibration for 2, 5 and 10 minutes, the survival rate of embryos which developed to normal blastocyst following thawing and culture processing were 58.5, 46.7 and 22.4%, respectively. The optimal time of equilibration of mouse morula and blastocysts in EFS solution seemed o be 2 and 5 ruin.

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Biomass Production and Phosphorus Inflow in three Perennial Herb Populations in the Basin of the Mt. Geumoh (금오산분지의 삼종 다년생 초목식물 개체군의 식물량생산과 인의 유입)

  • 유승원
    • Journal of Plant Biology
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    • v.29 no.2
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    • pp.95-107
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    • 1986
  • Seasonal changes in pool size, inflow rates in biomass and phosphorus, and the efficiency of phosphorus use in the stand of three populations (Helianthus tuberosus, Artemisia princeps and Phalaris arundinacea) in the basin of the Mt. Geumoh were investigated. During the early growing period, in the three species populations the relative size of the phosphorus pool of population was larger then that of its biomass pool, but that of the phosphorus pool of belowground part decreased more rapidly than that of its biomass pool. In the A. princeps and P. arundinacea populations, the phosphorus inflow rate was markedly high during the soil thaw in early spring and its seasonal change pattern was different from that of the biomass production rate, showing two peaks in March and June. But in the H. tuberosus population, the two seasonal change patterns were alike. The annual biomass production was 2283 gDM m-2 in the H. tuberosus, 1884 m-2 in the A. princeps and 1879 gDM m-2 in the P. arundinacea population, and the annual phosphorus inflow was 11.35, 9.63 and 7.60 gP m-2, respectively. The P. arundinacea population showed the smallest LAI peak(5.4 in early June), and the largest NAR peak (36.9 gDM m-2wk-1) RGR peak (0.15g g-1 wk-1) among the three species populations. The seasonal change patterns in whole plant EPU of the three species populations showed the bell shape, but the annual EPU values among them were markedly different. It was noticed that the population with the highest RGR showed the highest EPU among the three species populations while the population with the lowest RGR showed the lowest EPU among them.

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Effect of Short-term and Long-term Preservation on Motion Characteristics of Garole Ram Spermatozoa: A Prolific Microsheep Breed of India

  • Joshi, Anil;Bag, Sadhan;Naqvi, S.M.K.;Sharma, R.C.;Rawat, P.S.;Mittal, J.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.11
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    • pp.1527-1533
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    • 2001
  • Garole is a prolific, rare, less known and small size Indian sheep breed found in low and humid Sunderban region of West Bengal. Although information on stored Garole ram liquid semen upto 24 h is available, but there is a need to further investigate the short-term and long-term preservability of Garole ram semen for extensive utilization of this valuable germplasm by artificial insemination. The aim of the present study was to apply computer-assisted sperm analysis technique for assessing the motion characteristics of Garole ram semen stored (i) in liquid state at refrigeration temperature for short-term preservation upto 48 h and (ii) in frozen state at $-196^{\circ}C$ for long-term preservation after packaging in mini straws. Short-term preservation had a significant effect on motility (p<0.01) as the motility progressively decreased from 90.1% at 0 h to 85.5% and 73.2% after 24 and 48 h of storage, respectively. Although the decline in rapid moving sperms was also significant (p<0.01) on storage but the decrease was more pronounced at 48 h as compared to 24 h of storage period. Storage of chilled semen had also a significant effect on % linearity (p<0.05), % straightness (p<0.01), sperm velocities (p<0.01), amplitude of lateral head displacement (p<0.01) and beat frequency (pO.Ol) of spermatozoa. The replication had a significant effect for all the variables except average path and straight line velocity. However, the interactions of short-term storage and replication were non-significant for most of the variables except % of medium moving sperms, sperm velocities and beat frequency. On long-term preservation of Garole ram spermatozoa under controlled conditions the mean post-thaw recovery of 70.4 and 71.4% motile spermatozoa was achieved having 48.8 and 48.9% of rapidly motile spermatozoa, respectively in both the replicates. The effect of replication on cryopreservation was significant (p<0.05) on amplitude of lateral head displacement and beat frequency, but there was no significant effect on motility, rapidly motile spermatozoa, linearity, straightness and sperm velocities of frozen-thawed spermatozoa. It can be concluded from these results that an average 70% motility can be achieved on storage of Garole ram semen in chilled liquid state upto 48 h or in liquid nitrogen after freezing under controlled conditions in straws. However, further studies are required to evaluate the fertility of short-term and long-term preserved Garole ram semen for extensive use of this prolific sheep breed.