Using size-selected gold clusters on graphene oxide films to aid cryo-transmission electron tomography alignment.
Using size-selected gold clusters on graphene oxide films to aid cryo-transmission electron tomography alignment.
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DOI：
10.1038/srep09234
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发表时间：
2015-03-18
影响因子：
4.6
通讯作者：
Palmer RE
中科院分区：
文献类型：
--
作者：
Arkill KP;Mantell JM;Plant SR;Verkade P;Palmer RE
通过在玻璃冰中拍摄一系列以不同角度倾斜的透射电子显微照片，可以实现对纳米级水中物体的三维重建：低温透射电子断层扫描。本文提出了一种新的倾斜系列精密对准方法。在非晶态碳网上沉积了直径分别为~2.7 nm(Au5 6 1±14)、~3.2 nm(Au92 3±2 2)和~4.3 nm(Au2 0 5 7±45)的金团簇.在插入冷冻和随后传输到低温传输电子断层扫描后，所得到的断层图像在自动采集期间具有良好的(去)聚焦和对准特性。当均匀分布的3.2 nm的金粒子被用作基准标记时，精细排列是准确的，通过烟草花叶病毒的重建证明了这一点。使用氧化石墨烯薄膜意味着基准标记不会干扰冰结合的样品，并且自动收集是一致的。使用预沉积的大小选定的簇意味着没有聚集和用户定义的浓度。所选尺寸的团簇是单分散的，并且可以在包括直径2-5 nm的宽尺寸范围内产生。在氧化石墨烯薄膜上使用尺寸选择的团簇代表着3D冷冻电子显微镜的重大技术进步。
A three-dimensional reconstruction of a nano-scale aqueous object can be achieved by taking a series of transmission electron micrographs tilted at different angles in vitreous ice: cryo-Transmission Electron Tomography. Presented here is a novel method of fine alignment for the tilt series. Size-selected gold clusters of ~2.7 nm (Au561 ± 14), ~3.2 nm (Au923 ± 22), and ~4.3 nm (Au2057 ± 45) in diameter were deposited onto separate graphene oxide films overlaying holes on amorphous carbon grids. After plunge freezing and subsequent transfer to cryo-Transmission Electron Tomography, the resulting tomograms have excellent (de-)focus and alignment properties during automatic acquisition. Fine alignment is accurate when the evenly distributed 3.2 nm gold particles are used as fiducial markers, demonstrated with a reconstruction of a tobacco mosaic virus. Using a graphene oxide film means the fiducial markers are not interfering with the ice bound sample and that automated collection is consistent. The use of pre-deposited size-selected clusters means there is no aggregation and a user defined concentration. The size-selected clusters are mono-dispersed and can be produced in a wide size range including 2–5 nm in diameter. The use of size-selected clusters on a graphene oxide films represents a significant technical advance for 3D cryo-electron microscopy.
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影响因子：
48
作者：
Russo, Christopher J.;Passmore, Lori A.
通讯作者：
Passmore, Lori A.
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作者：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
Palmer, RE
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作者：
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通讯作者：
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