Journal
MICROSCOPY AND MICROANALYSIS
Volume 16, Issue 1, Pages 54-63Publisher
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S1431927609991280
Keywords
three-dimensional electron microscopy; aberration-corrected STEM; biological electron microscopy; thin sections; cytoskeleton; clathrin-coated pit; deconvolution; nanoparticles
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Funding
- Oak Ridge National Laboratory
- U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy, Vehicle Technologies Program
- Vanderbilt University Medical Center, National Institutes of Health [R01GM081801]
- NICHD
- U.S. Department of Energy Office of Science, Division of Materials Science and Engineering
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A three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM). The specimen was a metallic replica of the biological structure comprising Pt nanoparticles 2-3 nm in diameter, with a high stability under electron beam radiation. The 3D dataset was processed by an automated deconvolution procedure. The lateral resolution was 1.1 nm, set by pixel size. Particles differing by only 10 nm in vertical position were identified as separate objects with greater than 20% dip in contrast between them. We refer to this value as the axial resolution of the deconvolution or reconstruction, the ability to recognize two objects, which were unresolved in the original dataset. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy. However, the focal-series method does not require mechanical tilting and is therefore much faster. 3D STEM images were also recorded of the Golgi ribbon in conventional thin sections containing 3T3 cells with a comparable axial resolution in the deconvolved dataset.
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