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Three-Dimensional Electron Microscopy of Macromolecular AssembliesVisualization of Biological Molecules in Their Native State$
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Joachim Frank

Print publication date: 2006

Print ISBN-13: 9780195182187

Published to Oxford Scholarship Online: April 2010

DOI: 10.1093/acprof:oso/9780195182187.001.0001

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Electron Microscopy of Macromolecular Assemblies

Electron Microscopy of Macromolecular Assemblies

Chapter:
(p.15) 2 Electron Microscopy of Macromolecular Assemblies
Source:
Three-Dimensional Electron Microscopy of Macromolecular Assemblies
Author(s):

Joachim Frank

Publisher:
Oxford University Press
DOI:10.1093/acprof:oso/9780195182187.003.0002

Following an outline of the transmission electron microscope (TEM) and its working principles, this chapter starts with a description of specimen preparation methods for EM imaging, including negative staining, glucose embedment and ice embedment. The principle of image formation in the TEM is described as it pertains to biological weak phase objects, and in the process, the contrast transfer function (CTF) is introduced. It shows that EM images are effectively projections of the Coulomb potential distribution of the biological object, convoluted with that function. The chapter closes by describing methods for the determination and computational correction of the CTF.

Keywords:   contrast transfer function, Coulomb potential, ice embedment, glucose embedment, negative staining, phase object, transmission electron microscope

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