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 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.
Department of Molecular and Cell Biology, University of Cape Town, South Africa; Electron Microscope Unit, University of Cape Town, South Africa. Electronic address: jeremydavidwoodward@gmail.com.
J D
JD
Woodward
Electron Microscopy (EMEZ), ETH, Zürich, Switzerland.
R A
RA
Wepf
D000199:Actins / Q000737:chemistry / Q000648:ultrastructure; D000465:Algorithms; D000818:Animals; D021621:Imaging, Three-Dimensional / Q000379:methods* / Q000706:statistics & numerical data; D046911:Macromolecular Substances / Q000737:chemistry* / Q000648:ultrastructure*; D008855:Microscopy, Electron, Scanning / Q000379:methods* / Q000706:statistics & numerical data; D046529:Microscopy, Electron, Transmission / Q000379:methods / Q000706:statistics & numerical data; D011817:Rabbits; D059629:Signal-To-Noise Ratio; D013048:Specimen Handling
D000199:Actins; D046911:Macromolecular Substances
Filamentous actin; Iterative helical real-space reconstruction; Macromolecular; Scanning electron microscopy; Signal-to-noise ratio; Three-dimensional reconstruction