Optical sectioning microscopy: cellular architecture in three dimensions.
Optical sectioning microscopy: cellular architecture in three dimensions.
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DOI：
10.1146/annurev.bb.13.060184.001203
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
D. Agard
中科院分区：
文献类型：
--
作者：
D. Agard
在三维空间中分析生物标本的能力代表了现代结构生物学的主要成就之一。除了最简单的重复结构，三维分析是理解复杂生物组装的关键先决条件。宏观分子X射线晶体学已经为蛋白质、核酸和病毒提供了原子分辨率的三维结构。事实上，我们关于单个生物分子结构与功能关系的大部分基本知识都来自这些研究。电子显微镜已成为研究细胞成分的主要工具。随着Klug及其同事（9、10、17）的图像重建方法的发展，现在以中等分辨率（720 A）在三维中研究二维晶体或高对称性物体是常规的。最近，一些工作人员（22）甚至开始从切割的切片中重建相对较大的非晶体物体（1000 A），尽管分辨率较低（50-100 A）。相对便宜的计算机和数字图像采集系统的出现，现在已经有可能从光学显微镜拍摄的图像的三维重建。三维数据是通过记录一系列的图像在不同的焦平面在整个标本（光学切片）。可以使用光学显微镜可用的大多数常见成像方法。每个记录的图像代表
The ability to analyze biological specimens in three dimensions represents one of the major achievements of modern structural biology. For all but the simplest repeating structures, three-dimensional analysis is a crucial prerequisite for understanding complex biological assemblies. Macro molecular X-ray crystallography has provided three-dimensional structures at atomic resolution for proteins, nucleic acids, and viruses. Indeed, most of our fundamental knowledge on the relation of structure to func tion of individual biological molecules has come from these studies. Elec tron microscopy has been a major tool for the study of cell components. With the development of imagc reconstruction methods by Klug and co workers (9, 10,17) it is now routine to study two-dimensional crystalline or high symmetry objects in three dimensions at moderate resolutions (720 A). Recently, several workers (22) have even begun to reconstruct relatively large and noncrystalline objects ( � 1000 A) from cut sections, although at somewhat lower resolution's (�50-100 A). The advent of relatively inexpensive computers and digital image acquisition systems has now made possible the three-dimensional reconstruction of images taken from the optical microscope. Three-dimensional data is collected by recording a series of images taken at different focal planes throughout the specimen (optical sectioning). Most of the common imaging methods available with light microscopy may be used. Each recorded image represents the sum of