Cellular and Molecular Biotechnology Research Institute, Department of Life Science and Biotechnology, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central-6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan.
Electron Microscopy Facility, Open Research Facilities Station, Open Research Platform Unit, Tsukuba Innovation Arena (TIA) Central Office, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central-6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan.
Microsc Microanal. 2023 Dec 21;29(6):2080-2089. doi: 10.1093/micmic/ozad107.
Transmission electron microscopy (TEM) has revolutionized our understanding of protein structures by enabling atomic-resolution visualization without the need for crystallography, thanks to advancements in cryo-TEM and single particle analysis methods. However, conventional electron microscopy remains relevant for studying stained samples, as it allows the practical determination of optimal conditions through extensive experimentation. TEM also facilitates the examination of supramolecular complexes encompassing proteins, lipids, and nucleic acids. In this study, we investigated the applicability of lanthanoid reagents as electron-staining alternatives to uranyl acetate, which is globally regulated as a nuclear fuel material. We focus on a model biomembrane vesicle system, the chromatophores from the purple photosynthetic eubacterium Rhodospirillum rubrum, which integrate proteins and lipids. Through density distribution analysis of electron micrographs, we evaluated the efficacy of various lanthanoid acetates and found that triacetates of neodymium, samarium, and gadolinium exhibited similar staining effectiveness to uranyl acetate. Additionally, triacetates of praseodymium, erbium, and lutetium, followed by europium show promising results as secondary candidates. Our findings suggest that lanthanoid transition heavy metal acetates could serve as viable alternatives for electron staining in TEM, offering potential advantages over uranyl acetate.
透射电子显微镜(TEM)通过在无需结晶学的情况下实现原子分辨率的可视化,彻底改变了我们对蛋白质结构的理解,这要归功于低温 TEM 和单颗粒分析方法的进步。然而,传统的电子显微镜在研究染色样本时仍然具有相关性,因为它可以通过广泛的实验来确定最佳条件。TEM 还便于检查包含蛋白质、脂质和核酸的超分子复合物。在这项研究中,我们研究了镧系元素试剂作为铀酰醋酸盐的电子染色替代物的适用性,铀酰醋酸盐作为核燃料材料在全球范围内受到监管。我们专注于一个模型生物膜囊泡系统,即来自紫色光合作用细菌红假单胞菌的类囊体,该系统整合了蛋白质和脂质。通过电子显微镜照片的密度分布分析,我们评估了各种镧系元素醋酸盐的效果,并发现钕、钐和钆的三醋酸盐与铀酰醋酸盐具有相似的染色效果。此外,镨、铒和镥的三醋酸盐以及随后的铕显示出作为二级候选物的有希望的结果。我们的研究结果表明,镧系元素过渡重金属醋酸盐可用作 TEM 中电子染色的可行替代品,与铀酰醋酸盐相比具有潜在优势。