Abstract
Scanning electron microscopy (SEM) is a powerful technique that can image exposed surfaces in 3D. Modern scanning electron microscopes, with field emission electron sources and in-lens specimen chambers, achieve resolutions of better than 0.5 nm and thus offer views of ultrastructural details of subcellular structures or even macromolecular complexes. Obtaining a reliable image is, however, dependent on sample preparation methods that robustly but accurately preserve biological structures. In plants, exposing the object of interest may be difficult due to the existence of a cell wall. This protocol shows how to isolate plant nuclei for SEM imaging of the nuclear envelope and associated structures from both sides of the nuclear envelope in cultured cells as well as in leaf or root cells. Further, it provides a method for uncovering membrane-associated cytoskeletal structures.
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Acknowledgments
This work was supported by grants from the Biotechnology and Biological Sciences Research Council, UK, grant number BB/E015735/1 and BB/G011818/1. Thanks to Christine Richardson and Helen Grindley for technical assistance.
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Fišerová, J., Goldberg, M.W. (2014). Imaging Plant Nuclei and Membrane-Associated Cytoskeleton by Field Emission Scanning Electron Microscopy. In: Žárský, V., Cvrčková, F. (eds) Plant Cell Morphogenesis. Methods in Molecular Biology, vol 1080. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-643-6_14
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DOI: https://doi.org/10.1007/978-1-62703-643-6_14
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-642-9
Online ISBN: 978-1-62703-643-6
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