Focused-ion-beam thinning of frozen-hydrated biological specimens for cryo-electron microscopy
Focused-ion-beam thinning of frozen-hydrated biological specimens for cryo-electron microscopy
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DOI：
10.1038/nmeth1014
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发表时间：
2007-03-01
期刊：
影响因子：
48
通讯作者：
Mannella, Carmen
中科院分区：
文献类型：
--
作者：
Marko, Michael;Hsieh, Chyongere;Mannella, Carmen
低温电子显微镜可以提供有关细胞和细胞器在接近自然的冷冻水合状态下的高分辨率结构信息。然而，由于在制备适当薄的、玻璃化冷冻的生物标本时遇到的困难，其适用性受到限制。我们通过对大肠杆菌细胞的冷冻电子断层扫描，证明了聚焦离子束(FIB)可以用一种方便且基本上没有伪影的方式来稀释整个冷冻水合细胞。
Cryo-electron microscopy can provide high-resolution structural information about cells and organelles in the nearly native, frozen-hydrated state. Applicability, however, is limited by difficulties encountered in preparing suitably thin, vitreously frozen biological specimens. We demonstrate, by cryo-electron tomography of Escherichia coli cells, that a focused ion beam (FIB) can be used to thin whole frozen-hydrated cells in a convenient and essentially artifact-free way.