CRYOMICROSCOPIC OBSERVATIONS OF CATTLE EMBRYOS DURING FREEZING AND THAWING
CRYOMICROSCOPIC OBSERVATIONS OF CATTLE EMBRYOS DURING FREEZING AND THAWING
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DOI：
10.1016/0093-691x(83)90013-4
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发表时间：
1983-01-01
期刊：
影响因子：
2.8
通讯作者：
RALL, WF
中科院分区：
文献类型：
--
作者：
LEHNJENSEN, H;RALL, WF
使用冷冻显微镜观察在1.4 M甘油/PBS [磷酸盐缓冲盐水]中冷却和加温期间第6.5-7.5天牛胚胎外观的变化。胚胎以0.2 ℃至100 ℃的各种速率冷却。25度。以-25 ℃/分钟的速度升温至-25 ℃之间的温度。和-60 °。C，然后快速冷却（约250.degree. C/min）至低于-140 ℃的温度。C.在缓慢冷却过程中由横截面积计算的胚胎体积在-25 ℃时减小。C至等渗体积的约50%。在快速冷却至apprx后，在胚胎周围的细胞外冰基质中可以观察到断裂面。-140.degree. C.破裂面经常接触透明膜，有时会导致透明膜破裂。在从-20 ℃之间的温度快速冷却后，更经常观察到透明膜中的裂纹。至-35 °。C（9/13）比从-36 ℃之间的温度。至-60 °。C（2/7）。当胚胎迅速升温时（. apprx. 250.degree.温度低于-140 ℃/min）。C时，除细胞外冰融化外，胚胎外观及周围环境无变化。当胚胎缓慢升温（2.5 ℃）时，胚胎的发育受到抑制。或5度。C/min）时，观察到一系列事件; -70.degree. C细胞质和细胞外空间逐渐变暗，并在λ时达到最大暗度。-55.degree.然后，在持续缓慢升温的过程中，黑色物质逐渐消失;最后，大的细胞外冰晶融化了。
A cryomicroscope was used to observe changes in the appearance of day 6.5-7.5 cattle embryos during cooling and warming in 1.4 M glycerol/PBS [phosphate-buffered saline]. Embryos were cooled at various rates between 0.2.degree. and 25.degree. C/min to temperatures between -25.degree. and -60.degree. C, and then cooled rapidly (.apprx. 250.degree. C/min) to temperatures below -140.degree. C. The volume of the embryos calculated from the cross-sectional area during slow cooling decreased at -25.degree. C to about 50% of the isotonic volume. Fracture planes could be observed in the extracellular ice matrix surrounding the embryos after rapid cooling to .apprx. -140.degree. C. Fracture planes often touched the zona pellucida and sometimes caused cracks in the zona. Cracks in the zona pellucida were observed more often after rapid cooling from temperatures between -20.degree. to -35.degree. C (9/13) than from temperatures between -36.degree. to -60.degree. C (2/7). When embryos were warmed rapidly (.apprx. 250.degree. C/min) from temperatures below -140.degree. C, no change was observed in the appearance of the embryo or its surroundings except the melting of the extracellular ice. When embryos were warmed slowly (2.degree. or 5.degree. C/min), a series of events was observed; 1st, at .apprx. -70.degree. C the cytoplasm and the extracellular space gradually darkened and reached maximum darkness at .apprx. -55.degree. C; then, on continued slow warming, the dark material gradually disappeared; and finally the large extracellular ice crystals melted.