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首页> 外文期刊>Japanese Journal of Applied Physics. Part 1, Regular Papers, Brief Communications & Review Papers >Lipid Nanotube Encapsulating Method for Two- and Three-Dimensional Transmission Electron Microscopy Analyses of Cage-Shaped Proteins
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Lipid Nanotube Encapsulating Method for Two- and Three-Dimensional Transmission Electron Microscopy Analyses of Cage-Shaped Proteins

机译：笼形蛋白二维和三维透射电镜分析的脂质纳米管封装方法。

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摘要

A novel, sample-fixation technique for transmission electron microscopy (TEM) is proposed. We encapsulated various samples in one-dimensional hollow cylinders of a glycolipid nanotube (LNT) and found it to be highly stable, flexible, and reproducible. The cylindrical shape of the LNT allows us to observe nanomaterials consistently even when the samples are rotated around the tube axis. This is critical in improving TEM computed tomography (CT), which suffers from the loss of the electron beam at high tilting angles because of the increased effective thickness of the supporting film. This technique makes it possible to reconstruct three-dimensional images from weak-signal TEM images. In this work, we examined two types of cage-shaped proteins, ferritin, and DNA-binding protein from starved cells (Dps). We reconstructed three-dimensional images of Fe cores from zero-loss images or from electron energy-loss spectroscopy (EELS) mapping images obtained with the Fe M_(2,3) edge. We also applied negative staining and vitreous ice embedding (VIE). These results show the potential of the LNT encapsulation technique.
机译：提出了一种新颖的透射电子显微镜（TEM）的样品固定技术。我们将各种样品封装在糖脂纳米管（LNT）的一维空心圆柱体中，发现其高度稳定，灵活且可重现。 LNT的圆柱形状使我们能够一致地观察纳米材料，即使样品绕管轴旋转。这对于改善TEM计算机断层扫描（CT）是至关重要的，由于支撑膜的有效厚度增加，该技术在高倾斜角下会损失电子束。该技术使得从弱信号TEM图像重建三维图像成为可能。在这项工作中，我们检查了饥饿细胞（Dps）中的两种笼形蛋白，铁蛋白和DNA结合蛋白。我们从零损耗图像或电子能量损耗谱（EELS）映射图像（使用Fe M_（2,3）边缘获得）重建了Fe核的三维图像。我们还应用了负染和玻璃体冰埋（VIE）。这些结果表明了LNT封装技术的潜力。

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