牙鲆胚胎玻璃化冷冻及解冻过程中的温度因子与玻璃化率  

Study on temperature factor and vitrification rate in vitrification and thawing process of Japanese flounder embryos

作　　者：田永胜[1] 刘本伟[1] 陈松林[1] 

机构地区：[1]农业部海洋渔业资源可持续利用重点开放实验室中国水产科学研究院黄海水产研究所,青岛266071

出　　处：《海洋水产研究》2008年第5期59-64,共6页Marine Fisheries Research

基　　金：国家自然科学基金项目(30570259)资助

摘　　要：为避免鱼类胚胎玻璃化冷冻过程中脱玻璃化现象发生，本文对冷冻过程中温度变化及解冻方法进行了研究。在室温13℃下，将含有玻璃化液FVS3的麦管直接投入到-196℃的液氮中，麦管内温度达到-196℃时，所用的时间为15．46±1．38S，降温速率为800℃／min。对玻璃化麦管在35～45。C水浴中解冻时间进行测定。结果显示，解冻所需时间在6．17～7．94S之间，麦管从-196℃升高到13℃的解冻速率为1500～2000℃／min。当水浴温度从37℃逐步升高到72℃时，对水浴表面蒸气温度的变化进行了测定，相应水温下蒸气温度从25．47℃升高到38．6℃。在麦管中加入不同浓度的玻璃化液FVS1、FVS2、FVs3和FVS4进行玻璃化冷冻，然后将麦管分别在37℃水浴中和28．6℃的蒸气中解冻时玻璃化率进行研究。结果显示，在蒸气中解冻时的玻璃化率明显高于水浴中解冻；麦管玻璃化率在水浴中解冻时不稳定，在蒸气中解冻时表现稳定，玻璃化率随着玻璃化液浓度的升高有逐渐升高的趋势。对含有牙鲆胚胎的玻璃化冷冻麦管分别在37℃水浴和28．6℃蒸气中解冻，结果表明，蒸气中解冻时的玻璃化率同样明显高于水浴中解冻。In order to reduce the devitrification of fish embryo during the cryopreservation process, the aspects below were studied： temperature change inside the straws during freezing and thawing processes of vitrification, thawing time of straws in water bath and effect on the vitrification when the straws thawed in water bath and vapor respectively with and without eggs. When the straws loaded with vitrification solution FVS3 were plunged into liquid nitrogen at room temperature, it took 15.46 ± 1.38 seconds for the temperature fell to -196℃ from 13℃ inside the straws, and the cooling rate of the freezing process was 800℃/min. The thawing time was 6.17-7.94 seconds when the vitrified straws were thawed in water bath （37-45 ℃）, and the warming rate of the thawing process was 1 500-2 000℃/min. The temperature of the vapor at the surface of the water changed from 25.47℃ to 38.6℃ when the water temperature changed from 37℃ to 72℃. The vitrified straws loaded with different vitrification solutions FVS1 ,FVS2 ,FVS3 ,FVS4 were thawed in water bath （37 ℃） and vapor （28.6 ℃） respectively, and the vitrification rate was also studied. The result showed that the vitrification rate was higher when the straws were thawed in vapor than that in water bath and the process was stable when the straws were thawed in vapor while it was unstable in the water bath. The vitrification rate showed a trend of rising as the concentration of vitrification solution rose. The vitrified straws containing embryos of Japanese flounder Paralichthys olivaceus were also thawed in water bath and vapor at the temperatures mentioned above. Result showed that the vitrification rate was also higher when the straws were thawed in vapor than that in water bath.

关 键 词：牙鲆 胚胎 玻璃化冷冻 温度因子 玻璃化率 

分 类 号：Q65[生物学—生物物理学]