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Volume Scanning Electron Microscopy: Serial Block-Face Scanning Electron Microscopy Focussed Ion Beam Scanning Electron Microscopy

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Book cover Cellular Imaging

Part of the book series: Biological and Medical Physics, Biomedical Engineering ((BIOMEDICAL))

Abstract

Volume scanning electron microscopy is renewing interest in electron microscopy by offering three dimensional visualisation of biological systems. Two main contributors to this are the Serial Block-Face SEM (SBEM) and Focused Ion Beam SEM (FIB-SEM). The resolution offered by the FIB-SEM makes it the go-to tool for studies at higher resolution over smaller areas, particularly for subcellular studies. Compared to FIB-SEM, SBEM gives a much larger field of view (FOV) with a reduced resolution and anisotropic data. These methods require specific considerations as compared to the more traditional transmission electron microscopy. One major consideration is the sample preparation which is an integral part of the entire process for volume SEM. In this chapter we discuss the two techniques from a more practical approach.

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Acknowledgements

Thanks to the staff from both the EMBL Electron Microscopy Core Facility and the University of Queensland Centre for Microscopy and Microanalysis. The constant support from Yannick Schwab cannot be surpassed. We are also thankful to Anna Steyer and Rachel Mellwig for their helpful discussions and suggestions to the manuscript. A special mention to Matthia Karreman and José Miguel Serra Lleti. Without the work of Robyn Chapman we wouldn’t have a SBEM system and it’s her untiring efforts that have led to its development in our facilities. I thank her for all her determination to make this technique a success. Rob Parton has always been a source of encouragement for this work and has been willing to supply samples for us to work with when developing new ideas. Rachel Templin has been a wonderful backup helping us to process large numbers of samples as we have sought to develop new processing for this technology.

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Authors and Affiliations

  1. Centre for Microscopy and Microanalysis, The University of Queensland, Brisbane, QLD, 4072, Australia

    Richard I. Webb

  2. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117, Heidelberg, Germany

    Nicole L. Schieber

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  1. Richard I. Webb
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  2. Nicole L. Schieber
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Correspondence to Richard I. Webb .

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  1. Bio21 Institute Advanced Microscopy Facility, University of Melbourne, Melbourne, Victoria, Australia

    Eric Hanssen

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Webb, R.I., Schieber, N.L. (2018). Volume Scanning Electron Microscopy: Serial Block-Face Scanning Electron Microscopy Focussed Ion Beam Scanning Electron Microscopy. In: Hanssen, E. (eds) Cellular Imaging. Biological and Medical Physics, Biomedical Engineering. Springer, Cham. https://doi.org/10.1007/978-3-319-68997-5_5

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