Macromolecular 3D SEM reconstruction strategies: Signal to noise ratio and resolution

Three-dimensional scanning electron microscopy generates quantitative volumetric structural data from SEM images of macromolecules. This technique provides a quick and easy way to define the quaternary structure and handedness of protein complexes. Here, we apply a variety of preparation and imaging...

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Bibliographic Details
Published in:Ultramicroscopy 2014-09, Vol.144, p.43-49
Main Authors: Woodward, J.D., Wepf, R.A.
Format: Article
Language:English
Subjects:
Actins - chemistry
Actins - ultrastructure
Algorithms
Animals
Filamentous actin
Handedness
Imaging
Imaging, Three-Dimensional - methods
Imaging, Three-Dimensional - statistics & numerical data
Iterative helical real-space reconstruction
Macromolecular
Macromolecular Substances - chemistry
Macromolecular Substances - ultrastructure
Microscopy, Electron, Scanning - methods
Microscopy, Electron, Scanning - statistics & numerical data
Microscopy, Electron, Transmission - methods
Microscopy, Electron, Transmission - statistics & numerical data
Proteins
Rabbits
Reconstruction
Scanning electron microscopy
Signal to noise ratio
Specimen Handling
Strategy
Three dimensional
Three-dimensional reconstruction
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Summary:Three-dimensional scanning electron microscopy generates quantitative volumetric structural data from SEM images of macromolecules. This technique provides a quick and easy way to define the quaternary structure and handedness of protein complexes. Here, we apply a variety of preparation and imaging methods to filamentous actin in order to explore the relationship between resolution, signal-to-noise ratio, structural preservation and dataset size. This information can be used to define successful imaging strategies for different applications. •F-actin SEM datasets were collected using 8 different preparation/ imaging techniques.•Datasets were reconstructed by back projection and compared/analyzed•3DSEM actin reconstructions can be produced with
ISSN:0304-3991
1879-2723
DOI:10.1016/j.ultramic.2014.04.009