• Title/Summary/Keyword: Electron microscope grid

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마우스 성숙난자의 유리화동결법에 따른 동결 융해후의 염색체와 방추사의 분석

  • 박성은;신태은;김승범;차수경;임정묵;정형민
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.75-75
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    • 2001
  • 유리화 동결법은 동결중 ice crystal의 형성이 이루어지지 않으므로 난자의 동결보존을 위한 유용한 동결방법이다. 이전의 연구에서 마우스의 난자를 ethylene glycol과 electron microscope grid를 이용한 유리화 동결법으로 동결 융해한 결과 기존의 slow freezing 방법에서보다 높은 생존율과 배발달율이 나타남을 관찰하였다. 그러나 동물과 인간 난자를 이용한 연구를 통하여 난자의 경우 동결 융해후 염색체에 부착되어 있는 미세소관인 방추사가 온도의 변화에 매우 민감하여 염색체 이상성이 증가되는 것이 보고되었다. 이에 본 연구에서는 성숙난자를 유리화동결법에 의해 동결 융해후 난자의 염색체와 방추사의 이상성이 증가되는지 알아보고자 본 실험을 수행하였다. ICR mouse의 성숙란을 채취하여 연구목적에 따라 fresh한 대조군과 동결음해 시킨 실험군으로 분류하였다. 동결방법은 난자를 1.5 M ethylene glycol (EG)에 2분 30초간 노출 시킨후 5.5 M EG와 1M sucrose가 첨가된 동결액에 20초간 노출시킨 후 Grid에 난자를 부착시킨 후 직접 액체질소에 침지하여 동결하였다. 동결후 난자는 5단계로 융해를 실시한 후 생존된 난자는 tubulin 항체를 이용한 immunostaining 방법으로 방추사의 이상성을 관찰하였고, 염색체는 난자를 고정 후 10% Giemza로 염색 후 염색체의 수적인 이상성을 관찰하였다. 염색체 분석결과 염색체 이상 빈도는 대조군의 경우 19.6%, 동결융해군은 32.8%로 관찰되었다. 또 방추사의 이상빈도는 대조군의 경우 20.2%, 동결 융해군은 32.3%로 관찰되어 동결 융해후의 난자에서 염색체와 방추사의 이상 빈도가 증가됨이 관찰되었다.찰되었다. 배아이식후 대조군과 실험군에서 각각 2 마리가 임신이되어 정상적인 산자를 분만하였다. 따라서 항동해제에 taxol의 첨가는 동결 융해후의 난자의 배발달율을 증진시킬 수 있었다..8%로 나타나 난할율 및 배반포 발생율에 있어서 융합조건에 따라 큰 차이는 없었으나 1.9㎸/cm, 30$\mu\textrm{s}$ 2회의 조건이 다른 조건들에 비하여 유의적으로 낮았다. 따라서, 체세포와 수핵란 세포질간의 융합율과 배반포 발생에 미치는 영향은 전압보다는 시간에 더 크게 받음을 알 수 있었으며, 이와 같은 결과에서 융합시 시간을 오래 주는 것보다 전압을 높이는 것이 수핵난자의 세포질에 상해를 줄이고 이후 배반포 발생에 유리할 것으로 사료되었다.면에서도 더욱 더 활발할 것으로 기대된다. 배란후 72시간째에 초음파진단기를 이용하여 난소의 난포발달을 조사한 결과 , 대조구와 bFF처리구에 비해 AI처리구에서 발달난포가 유의적으로 많은 것을 확인하였다. 이상과 같은 결과로, Anti-inhibin serum은 한우 자체에서 분비하는 Inhibin을 특이하게 억제하여 Inhibin에 의해 억제되는 FSH분비가 촉진됨으로써 난포발달과 estrogen의 농도가 촉진되는 것으로 사료되어 anti-inhibin serum이 한우의 과배란유기 효과가 있는 것으로 사료된다.정량 분석한 결과이다. 시편의 조성은 33.6 at% U, 66.4 at% O의 결과를 얻었다. 산화물 핵연료의 표면 관찰 및 정량 분석 시험시 시편 표면을 전도성 물질로 증착시키지 않고, Silver Paint 에 시편을 접착하는 방법으로도 만족한 시험 결과를 얻을 수 있었다.째, 회복기 중에 일어나는 입자들의 유입은 자기폭풍의 지속시간을 연장시키는 경향을 보이며

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In Vitro Fertilization and Development of Bovine Immature Oocytes by Ultra- Rapid Cooling (미성숙 소난자의 초급속 동결ㆍ융해 후 수정 및 체외 배발달)

  • Ryu, C. S.;K. S. Kil;Kim, N-H;D. H. Ko;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.23 no.3
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    • pp.221-227
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    • 1999
  • Successful cryopreservation of bovine immature oocytes can increase availably of oocytes for the in vitro fertilization or nuclear transfer. However, it was not reported successful development to the blastocyst stage following in vitro fertilization of cryopreserved bovine immature oocytes. The objective of this study was to determine the incidence of survival, meiotic maturation, fertilization and in vitro development of cryopreserved bovine immature by ultra rapid cooling methods. The oocytes were adversely affected by brief exposure to EFS40 solution in electron microscope grids and plunged directly into liquid nitrogen. After such ultra-rapid cooled immature oocytes were warmed, 78% of oocytes were matured to the metaphase II stage, 50% of oocytes were fertilized after insemination, and 5% of oocytes were developed to the blastocyst stage. Different sodium concentration of sodium ion in the freezing medium did not affect survival, maturation, fertilization and in vitro development of cryopreserved oocytes. These results suggested that immature bovine oocytes can be cryopreserved by ultra-rapid cooling methods.

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Characteristic of Intermetallic Compounds for Aging of Lead Free Solders Applied to 48 $\mu$BGA (48 $\mu$BGA에 적용한 무연솔더의 시효처리에 대한 금속간화합물의 특성)

  • Shin, Young-Eui;Lee, Suk;Fujimoto, Kozo;Kim, Jong-Min
    • Journal of the Microelectronics and Packaging Society
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    • v.8 no.3
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    • pp.37-42
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    • 2001
  • The concerns of the toxicity and health hazard of lead in solders have demanded the research to find suitable lead-free solder alloys. It was discussed that effect of the intermetallic formation and structure on the reliability of solder joints. In this study, lead-free solder alloys with compositions of Sn/3.5Ag/0.75Cu, Sn/2.0Ag/0.5Cu/2.0Bi were applied to the 48 $\mu$BGA packages. Also, the lead-free solder alloys compared with eutectic Sn/37Pb solder using shear test under various aging temperature. Common $\mu$BGA with solder components was aged at $130^{\circ}C$, $150^{\circ}C$ and $170^{\circ}C$. And the each temperature applied to 300, 600 and 900 hours. The thickness of the intermetallics was measured for each condition and the activation energy for their growth was computed. The fracture surfaces were analyzed using SEM (Scanning Electron Microscope) with EDS (Energy Dispersive Spectroscopy). These results for reliability of lead-free interconnections are discussed.

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Study on the Vitrification of Porcine GV and M II Oocytes after Removal of Cytoplasmic Lipid Droplets (세포질 내 지방구 제거가 돼지 난포란의 유리화 동결에 미치는 영향)

  • 최인경;이승진;송해범
    • Journal of Embryo Transfer
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    • v.18 no.1
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    • pp.1-14
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    • 2003
  • This study was conducted to investigate that the immature and mature oocytes of porcine can be cryopreserved by vitrification. Oocytes were centrifuged to polarize the cytoplasmic lipid droplets. The lipids were removed from cytoplasm by micromanipulation. Delipated oocytes were centrifuged after being preincubated with cytochalasin B(CB) fer 10 min, and lipid droplets were removed. Centrifuged oocytes were treated with CB and centrifuged to polorize lipid droplets but not delipated and control oocytes is not-treatment. Oocytes of three types were vitrified in electron microscope(EM) grids. The results of survival, maturation and cleavage rates were as follows. 1 The survival rates of immature oocytes were 15.1%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated wassignificantly higher than Centrifuged and Control(P<.01). 2. The survival rates of mature oocytes were 12.21%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated was significantly higher than Centrifuged and Control(P<.01). 3 The maturation rates of immature oocytes were 37.5% and 68.9% for metaphase II in the Delipated after vitrification and Non-vitrification, respectively, and its rate of Non-vitrification was significantly higher than Delipated after vitrification(P<.01). 4. The cleavage rates of immature oocytes were 12.5%, 0%, 0% and 56.1% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05). 5 The cleavage rates of mature oocytes were 25.0%, 0%, 0% and 67.9% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05).