【摘要】 冷冻电镜是发现、鉴定病原微生物，解析病原微生物结构的有力工具。对冷冻电镜的实验过程中，样本是否会扩散到环境中导致污染，继而引发生物安全的问题，国内外目前都缺少相关实验探究。本研究利用当前广泛使用的直接投入式冷冻电镜样品制备设备，对病毒和细菌样品进行冷冻电镜样品制备，探究冷冻电镜制样过程中可能出现的样品泄露以及造成的污染问题，进行生物安全评估。实验结果表明：被滤纸吸附的样品可透过滤纸进一步污染制样设备，使用封口膜可有效避免样品进一步污染；病毒性样品实验结果表明即使用封口膜也可由多种原因而扩大污染范围。这些结果为冷冻电镜样品制备生物安全评估提供了实验依据。更多还原

【Abstract】 Cryo-electron microscopy(Cryo-EM) is a powerful tool for discovering and identifying pathogenic microorganisms and determining their structures. There is a lack of relevant research on whether the samples may spread to the environment and cause potential biosafety risks during the vitrification process of Cryo-EM sample preparation. In order to probe whether samples will cause contamination during Cryo-plunge, we chose biologically safe microorganisms such as Escherichia coli and Autographa californica multiple nucleopolyhedrovirus(AcMNPV)as the model samples and detected samples from different locations of the Cryo-plunge device and environment during the rapid cooling. The result showed that bacteria sample could be detected on the filter paper which has no direct contact with the sample and the replacement with a parafilm can block the pollution. Surprisingly, the virus sample can cause further penetrate parafilm and incur contamination probably due to multi-factors. Our result provide an experimental basis for bio-safety assessment of Cryo-EM sample preparation and shed light on the construction of higher bio-safety level lab with Cryo-EM facility.更多还原