Negative staining and image classification - Powerful tools in modern electron microscopy

被引：516

作者：

Ohi M. ^{[1
]}

Li Y. ^{[2
]}

Cheng Y. ^{[1
]}

Walz T. ^{[1
]}

机构：

[1] Department of Cell Biology, Harvard Medical School, Boston, MA 02115

[2] Dept. Biol. Chem./Molec. Pharmacol., Harvard Medical School, Boston, MA 02115

来源：

Biological Procedures Online | 2004年 / 6卷 / 1期

关键词：

Electron; Microscopy; Negative staining; Protein conformation;

D O I：

10.1251/bpo70

中图分类号：

学科分类号：

摘要：

Vitrification is the state-of-the-art specimen preparation technique for molecular electron microscopy (EM) and therefore negative staining may appear to be an outdated approach. In this paper we illustrate the specific advantages of negative staining, ensuring that this technique will remain an important tool for the study of biological macromolecules. Due to the higher image contrast, much smaller molecules can be visualized by negative staining. Also, while molecules prepared by vitrification usually adopt random orientations in the amorphous ice layer, negative staining tends to induce preferred orientations of the molecules on the carbon support film. Combining negative staining with image classification techniques makes it possible to work with very heterogeneous molecule populations, which are difficult or even impossible to analyze using vitrified specimens. © 2004. Biological Procedures Online.

引用

页码：23 / 34

页数：11

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