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Journal of Cell Science 115, 1877-1882 (2002)
© 2002 The Company of Biologists Limited

Research Article

Visualisation of the actin cytoskeleton by cryo-electron microscopy

Guenter P. Resch1, Kenneth N. Goldie2, Angelika Krebs2, Andreas Hoenger2 and J. Victor Small*,1

1 Institute of Molecular Biology, Department Cell Biology, Billrothstrasse 11, A-5020 Salzburg, Austria
2 European Molecular Biology Laboratory, Structure Programme, Meyerhofstrasse 1, D-69117 Heidelberg, Germany

* Author for correspondence (e-mail: jvsmall{at}imb.oeaw.ac.at )

Accepted 12 February 2002

An understanding of the mechanisms driving cell motility requires clarification of the structural organisation of actin filament arrays in the regions of cell protrusion termed lamellipodia. Currently, there is a lack of consensus on lamellipodia organisation stemming from the application of alternative procedures for ultrastructural visualisation of cytoskeleton networks. In this study, we show that cryo-electron microscopy of extracted cytoskeletons embedded in a thin layer of vitreous ice can reveal the organisation of cytoskeletal elements at high resolution. Since this method involves no dehydration, drying and contrasting steps that can potentially introduce subtle distortions of filament order and interactions, its application opens the way to resolving the controversial details of lamellipodia architecture.

Key words: Actin, Cytoskeleton, Lamellipodium, Motility, Cryo-electron microscopy


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