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Automatic Acquisition and Image Analysis of 2D Crystals

Published online by Cambridge University Press:  14 March 2018

N. Coudray*
Affiliation:
MIPS Laboratory, Université de Haute Alsace, Mulhouse, France
F. Beck
Affiliation:
MPIB, Max Planck Institute of Biochemistry, Molecular Structural Biology, Martinsried, Germany
J.-L. Buessler
Affiliation:
MIPS Laboratory, Université de Haute Alsace, Mulhouse, France
A. Korinek
Affiliation:
MPIB, Max Planck Institute of Biochemistry, Molecular Structural Biology, Martinsried, Germany
A. Karathanou
Affiliation:
MIPS Laboratory, Université de Haute Alsace, Mulhouse, France
H. Rémigy
Affiliation:
BIOZENTRUM, M.E. Mueller Institute for Structural Biology, University of Basel, Switzerland
H. Kihl
Affiliation:
MIPS Laboratory, Université de Haute Alsace, Mulhouse, France
A. Engel
Affiliation:
BIOZENTRUM, M.E. Mueller Institute for Structural Biology, University of Basel, Switzerland
J.M. Plitzko
Affiliation:
MPIB, Max Planck Institute of Biochemistry, Molecular Structural Biology, Martinsried, Germany
J.-P. Urban
Affiliation:
MIPS Laboratory, Université de Haute Alsace, Mulhouse, France

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2D-crystallization in combination with transmission electron microscopy (TEM) is one of the few methods for the structural analysis of membrane proteins in their native state. However, the parameters for the generation of large crystalline sheets are typically difficult to identify for a given protein. Many repetitive and time consuming screening steps by TEM are therefore necessary to find the best crystallization and preparation conditions. Although several software packages offer the possibility to control an electron microscope, none is completely adapted for a fully automated and completely integrated acquisition and analysis of 2D crystals. Here we report on the development of a fully automatic screening and on-line analysis software for the fast and automatic survey of large quantities of negatively stained EM samples for 2D crystallography.

Type
Research Article
Copyright
Copyright © Microscopy Society of America 2008

References

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[6]Acknowledgment: This work was supported by the EU 6th framework (HT- 3DEM, LSHG-CT-2005-018811).Google Scholar