【摘要】 负染技术在微生物的透射电镜观察中至关重要，但其伴随的生物安全问题亟待深入研究和探讨。关于负染制样过程中样品泄露的具体环节及其潜在风险，现有研究尚缺乏明确界定。本研究针对细菌与病毒样品，全面系统地评估了负染制样流程中可能存在的泄露风险点及其对生物安全的影响。结果表明，直接接触样品的器具构成了污染的主要来源，需特别加强防护与管理；透射电镜的真空环境及高能电子束的辐照作用均展现出了显著的样品灭活效果，尤其是当二者协同作用时，效果更为显著，从而有效降低了生物安全风险。本研究不仅加深了对微生物样品负染技术生物安全性的认知，还为构建更加科学、严谨且安全的微生物样品负染操作规范提供了参考依据。更多还原

【Abstract】 The negative staining technique is essential for transmission electron microscopy(TEM) observation of microorganisms. However, the biosafety concerns associated with this method require further urgent investigation and discussion. Current research provides limited clarity on the specific stages at which sample leakage occurs and the associated risks during the negative staining preparation process. This study comprehensively evaluated the potential leakage points and their impacts on biosafety during the negative staining preparation of bacterial and viral samples. The results indicated that items in direct contact with the samples were the primary source of contamination, highlighting the need for enhanced protection and management protocols. Furthermore, the vacuum environment of the TEM and the irradiation effects of high-energy electron beams were found to significantly inactivate the samples, particularly when acting synergistically, thereby reducing biosafety risks. This research not only enhances the understanding of biosafety in negative staining techniques for microbial samples but also provides a foundation for establishing more scientific, rigorous, and safe operational norms for the negative staining of microbial samples.更多还原