Towards high-resolution three-dimensional imaging of native mammalian tissue: Electron tomography of frozen-hydrated rat liver sections
Towards high-resolution three-dimensional imaging of native mammalian tissue: Electron tomography of frozen-hydrated rat liver sections
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DOI：
10.1016/j.jsb.2005.10.004
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发表时间：
2006-01-01
影响因子：
3
通讯作者：
Marko, M
中科院分区：
文献类型：
--
作者：
Hsieh, CE;Leith, A;Marko, M
冷冻水化标本的冷冻电子断层扫描具有相当大的希望，在其天然细胞环境中的细胞器和大分子复合物的高分辨率三维成像。虽然该技术已成功地用于小的，快速冷冻的细胞和细胞器，但应用于大量哺乳动物组织已被证明是困难的。我们报告了高压冷冻和冷冻超微切片制备的大鼠肝脏冷冻水化切片的冷冻电子断层扫描的进展。改进包括识别用于安装断层扫描切片的合适网格，减少切片上的表面伪影，通过使用能量过滤提高图像质量，以及使用活检针进行更快速的组织切除。冷冻水化肝切片的断层扫描重建显示了线粒体、内质网和核糖体等细胞成分的天然结构，而没有与冷冻置换使用的渗透和后染色相关的超微结构细节的选择性衰减或增强。(C)2005年爱思唯尔公司All rights reserved.
Cryo-electron tomography of frozen-hydrated specimens holds considerable promise for high-resolution three-dimensional imaging of organelles and macromolecular complexes in their native cellular environment. While the technique has been successfully used with small, plunge-frozen cells and organelles, application to bulk mammalian tissue has proven to be difficult. We report progress with cryo-electron tomography of frozen-hydrated sections of rat liver prepared by high-pressure freezing and cryo-ultramicrotomy. Improvements include identification of suitable grids for mounting sections for tomography, reduction of surface artifacts on the sections, improved image quality by the use of energy filtering, and more rapid tissue excision using a biopsy needle. Tomographic reconstructions of frozen-hydrated liver sections reveal the native structure of such cellular components as mitochondria, endoplasmic reticulum, and ribosomes, without the selective attenuation or enhancement of ultrastructural details associated with the osmication and post-staining used with freeze-substitution. (C) 2005 Elsevier Inc. All rights reserved.