Chapter 5
Volume Scanning Electron Microscopy:
Serial Block-Face Scanning Electron
Microscopy Focussed Ion Beam Scanning
Electron Microscopy
Richard I. Webb and Nicole L. Schieber
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.
5.1 Introduction
Gaining three-dimensional (3D) information about a biological system at the cellular level allows a better understanding of both its structure and function. In
biology, early electron microscopy relied on gaining ultrastructural information by
observing flat two dimensional ultrathin sections (50–60 nm) and trying to infer the
three dimensional structure from them. However, each section supplies only a
snapshot of an extremely small part of the cell, thus many things are overlooked and
wrong interpretations can be made.
R. I. Webb (&)
Centre for Microscopy and Microanalysis, The University of Queensland,
Brisbane, QLD 4072, Australia
e-mail: r.webb@uq.edu.au
N. L. Schieber
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL),
69117 Heidelberg, Germany
e-mail: nicole.schieber@embl.de
© Springer International Publishing AG 2018
E. Hanssen (ed.), Cellular Imaging, Biological and Medical Physics,
Biomedical Engineering, https://doi.org/10.1007/978-3-319-68997-5_5
117
Volume Scanning Electron Microscopy:
Serial Block-Face Scanning Electron
Microscopy Focussed Ion Beam Scanning
Electron Microscopy
Richard I. Webb and Nicole L. Schieber
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.
5.1 Introduction
Gaining three-dimensional (3D) information about a biological system at the cellular level allows a better understanding of both its structure and function. In
biology, early electron microscopy relied on gaining ultrastructural information by
observing flat two dimensional ultrathin sections (50–60 nm) and trying to infer the
three dimensional structure from them. However, each section supplies only a
snapshot of an extremely small part of the cell, thus many things are overlooked and
wrong interpretations can be made.
R. I. Webb (&)
Centre for Microscopy and Microanalysis, The University of Queensland,
Brisbane, QLD 4072, Australia
e-mail: r.webb@uq.edu.au
N. L. Schieber
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL),
69117 Heidelberg, Germany
e-mail: nicole.schieber@embl.de
© Springer International Publishing AG 2018
E. Hanssen (ed.), Cellular Imaging, Biological and Medical Physics,
Biomedical Engineering, https://doi.org/10.1007/978-3-319-68997-5_5
117