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对虾白斑综合症病毒透射电镜分析中病毒固定条件的优化

Optimization of white spot syndrome virus fixation method for transmission electron microscopy analysis
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摘要 电镜观察是研究对虾白斑综合症病毒(WSSV)形态和结构的重要手段.本研究采用不同浓度的多聚甲醛和戊二醛对纯化的WSSV粒子进行固定,从中寻找最优的固定条件用于病毒的透射电镜(TEM)分析.实验结果显示,经过多聚甲醛或者戊二醛固定的WSSV粒子对吸附、染色等实验操作的耐受性较未固定样品有显著提高,在制样过程中没有发生明显的结构破坏.其中1.00%(V/V)多聚甲醛常温固定20 min对WSSV粒子的结构保持得最好.进一步地分析表明,1.00%多聚甲醛固定后的病毒粒子更有利于金标免疫电镜的研究. Electron microscopic observation is an important method for morphology analysis of white spot syndrome virus（WSSV）. In this study, to optimize the fixation method, purified WSSV virions were treated with different concentration of paraformaldehyde or glutaraldehyde. The results showed that, paraformaldehyde and glutaraldehyde treatment could help to preserve the structure of WSSV in the process of transmission electron microscopy sample preparation. Fixation with 1.00% paraformaldehyde for 20 minutes had yielded the best result. Further analysis showed that virions fixed under this condition was suitable for immune electron microscopy analysis.
作者 邬君君 谷力 李钫
机构地区 国家海洋局第三海洋研究所
出处 《应用海洋学学报》 CAS CSCD 2013年第2期283-287,共5页 Journal of Applied Oceanography
基金 国家自然科学基金资助项目(40976100) 国家973计划资助项目(2012CB114401) 国家海洋局第三海洋研究所基本科研业务费专项资金资助项目(海三科2009044)
关键词 海洋生物学 固定 对虾白斑综合症病毒 多聚甲醛 戊二醛 marine biology fix white spot syndrome virus paraformaldehyde glutaraldehyde
分类号 P735 [天文地球—海洋生物学]
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参考文献11

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应用海洋学学报
2013年 第2期

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