Chapter 1
Electron Tomography: A Primer
Andrew Leis
Abstract This chapter is an introduction to the theory and practise of electron
tomography. It identifies the areas in need of most attention to maximise the efficiency of descriptive studies of morphology, and in the case of structural biology, to
enable the transition from a reductionist approach to in-depth systems biology. The
chapter concludes with a step-by-step guide to acquiring tomograms.
1.1 An Introduction to Electron Tomography
Electron tomography provides the unique possibility to view three-dimensional (3D),
nanometre-scale detail within the cellular context (for detailed reviews, see [1–3]). It is
one of a small group of bioimaging technologies that utilise electrons or photons to
deliver 3D information about structural complexity spanning macromolecules,
viruses, cells and tissues. These techniques include confocal microscopy, some forms
of super-resolution microscopy, and soft X-ray tomography, albeit with differing
means of generating contrast, and overlapping resolution. With proper attention to
structural preservation, these techniques are capable of capturing and deciphering
cellular events such as the workings of protein machines in healthy cells [4] and in
disease, a special case being the morphogenesis of infectious agents [5–8]. The
common feature of these techniques is the ability to image hydrated (or
frozen-hydrated) specimens. Confocal and super-resolution microscopy techniques
visualise fluorescent ‘spots’ rather than actual densities but they also provide unambiguous identification and possibly some temporal resolution. Furthermore, the spatial information can be decisive, as is the case for fluorescence (‘Förster’) resonance
energy transfer (FRET) analysis of protein-protein interactions [9]. Cryo- electron
tomography captures snapshots of true densities within the cellular landscape but
given the prospects for macromolecular crowding, especially in prokaryotes [10, 11]
A. Leis (&)
CSIRO Australian Animal Health Laboratory,
Private Bag 24, Geelong, VIC 3222, Australia
e-mail: leisandrew@gmail.com
© 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_1
1
Electron Tomography: A Primer
Andrew Leis
Abstract This chapter is an introduction to the theory and practise of electron
tomography. It identifies the areas in need of most attention to maximise the efficiency of descriptive studies of morphology, and in the case of structural biology, to
enable the transition from a reductionist approach to in-depth systems biology. The
chapter concludes with a step-by-step guide to acquiring tomograms.
1.1 An Introduction to Electron Tomography
Electron tomography provides the unique possibility to view three-dimensional (3D),
nanometre-scale detail within the cellular context (for detailed reviews, see [1–3]). It is
one of a small group of bioimaging technologies that utilise electrons or photons to
deliver 3D information about structural complexity spanning macromolecules,
viruses, cells and tissues. These techniques include confocal microscopy, some forms
of super-resolution microscopy, and soft X-ray tomography, albeit with differing
means of generating contrast, and overlapping resolution. With proper attention to
structural preservation, these techniques are capable of capturing and deciphering
cellular events such as the workings of protein machines in healthy cells [4] and in
disease, a special case being the morphogenesis of infectious agents [5–8]. The
common feature of these techniques is the ability to image hydrated (or
frozen-hydrated) specimens. Confocal and super-resolution microscopy techniques
visualise fluorescent ‘spots’ rather than actual densities but they also provide unambiguous identification and possibly some temporal resolution. Furthermore, the spatial information can be decisive, as is the case for fluorescence (‘Förster’) resonance
energy transfer (FRET) analysis of protein-protein interactions [9]. Cryo- electron
tomography captures snapshots of true densities within the cellular landscape but
given the prospects for macromolecular crowding, especially in prokaryotes [10, 11]
A. Leis (&)
CSIRO Australian Animal Health Laboratory,
Private Bag 24, Geelong, VIC 3222, Australia
e-mail: leisandrew@gmail.com
© 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_1
1