A preparation of bacterial outer membrane with osmium tetroxide and uranyl acetate co-stain enables improved structural determination by transmission electron microscopy.

Aadil Sheikh, Bernd Zechmann, Christie M Sayes, Joseph H Taube, K Leigh Greathouse
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Abstract

Biological nanoparticles, such as bacterial outer membrane vesicles (OMVs), are routinely characterized through transmission electron microscopy (TEM). In this study, we report a novel method to prepare OMVs for TEM imaging. To preserve vesicular shape and structure, we developed a dual fixation protocol involving osmium tetroxide incubation prior to negative staining with uranyl acetate. Combining osmium tetroxide with uranyl acetate resulted in preservation of sub-50 nm vesicles and improved morphological stability, enhancing characterization of lipid-based nanoparticles by TEM.

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用四氧化锇和醋酸铀酰共染色剂制备细菌外膜,改进了透射电子显微镜的结构测定。
生物纳米颗粒,如细菌外膜囊泡(OMVs),通常通过透射电子显微镜(TEM)进行表征。在这项研究中,我们报告了一种制备用于TEM成像的omv的新方法。为了保持囊泡的形状和结构,我们开发了一种双重固定方案,包括四氧化锇培养,然后用醋酸铀酰进行阴性染色。四氧化二锇与醋酸铀酰的结合可以保留50 nm以下的囊泡,提高形态稳定性,并通过TEM增强了脂基纳米颗粒的表征。
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