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膜结合细胞器的电子断层扫描术。

Electron tomography of membrane-bound cellular organelles.

作者信息

Frey Terrence G, Perkins Guy A, Ellisman Mark H

机构信息

Department of Biology, San Diego State University, San Diego, California 92182-4614, USA.

出版信息

Annu Rev Biophys Biomol Struct. 2006;35:199-224. doi: 10.1146/annurev.biophys.35.040405.102039.

Abstract

Electron microscope tomography produces three-dimensional reconstructions and has been used to image organelles both isolated and in situ, providing new insight into their structure and function. It is analogous to the various tomographies used in medical imaging. Compared with light microscopy, electron tomography offers an improvement in resolution of 30- to 80-fold and currently ranges from 3 to 8 nm, thus filling the gap between high-resolution structure determinations of isolated macromolecules and larger-scale studies on cells and tissues by light microscopy. Here, we provide an introduction to electron tomography and applications of the method in characterizing organelle architecture that also show its power for suggesting functional significance. Further improvements in labeling modalities, imaging tools, specimen preparation, and reconstruction algorithms promise to increase the quality and breadth of reconstructions by electron tomography and eventually to allow the mapping of the cellular proteomes onto detailed three-dimensional models of cellular structure.

摘要

电子显微镜断层扫描可生成三维重建图像,已被用于对分离的细胞器和原位细胞器进行成像,为其结构和功能提供了新的见解。它类似于医学成像中使用的各种断层扫描技术。与光学显微镜相比,电子断层扫描的分辨率提高了30至80倍,目前范围为3至8纳米,从而填补了分离的大分子高分辨率结构测定与光学显微镜对细胞和组织进行的更大规模研究之间的空白。在这里,我们介绍电子断层扫描及其在表征细胞器结构方面的应用,这些应用也展示了该方法在揭示功能意义方面的强大作用。标记方式、成像工具、样本制备和重建算法的进一步改进有望提高电子断层扫描重建的质量和广度,并最终实现将细胞蛋白质组映射到细胞结构的详细三维模型上。

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