Chapter 3
Electron Cryo-Tomography
Josie L. Ferreira, Teige R. S. Matthews-Palmer and Morgan Beeby
Abstract Electron cryo-tomography is an integrative imaging technique that produces high-resolution 3-D images of cells and purified complexes. These images
can uniquely provide mechanistic insights into in situ biology at molecular and
cellular scales. To produce these images, specimens are preserved by flash-freezing
without stains or fixatives; these flash-frozen specimens are imaged over a range of
angles in an electron microscope and this data used to reconstruct 3-D images of the
specimen in near-native state to ‘macromolecular’ resolution. In this introductory
chapter we take the perspective of the workflow of a cryo-tomography project to
outline the technique, and important underlying concepts.
3.1 Introduction
Advances in biological sciences have been critically dependent upon the imaging
techniques available at the time. It follows that understanding the mechanistic
underpinnings of biology in the 21st century relies upon techniques capable of
visualizing the components of entire biological processes in situ, and the emergent
properties of their interactions.
A technique capable of providing such holistic biological insights is electron
cryo-tomography (ECT), an electron microscopy approach that images flash-frozen
specimens in three-dimensions, over scales from molecular to cellular, to resolutions capable of discerning individual macromolecular complexes. Because the
specimen is cryo-preserved without stains, tomographic images are derived from
phase-contrast with the biological material itself and therefore reveal the molecular
J. L. Ferreira Á T. R. S. Matthews-Palmer Á M. Beeby (&)
Department of Life Sciences, Imperial College London, SW7 2AZ London, UK
e-mail: mbeeby@imperial.ac.uk
J. L. Ferreira
e-mail: j.ferreira14@imperial.ac.uk
T. R. S. Matthews-Palmer
e-mail: t.matthews-palmer14@imperial.ac.uk
© 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_3
61
Electron Cryo-Tomography
Josie L. Ferreira, Teige R. S. Matthews-Palmer and Morgan Beeby
Abstract Electron cryo-tomography is an integrative imaging technique that produces high-resolution 3-D images of cells and purified complexes. These images
can uniquely provide mechanistic insights into in situ biology at molecular and
cellular scales. To produce these images, specimens are preserved by flash-freezing
without stains or fixatives; these flash-frozen specimens are imaged over a range of
angles in an electron microscope and this data used to reconstruct 3-D images of the
specimen in near-native state to ‘macromolecular’ resolution. In this introductory
chapter we take the perspective of the workflow of a cryo-tomography project to
outline the technique, and important underlying concepts.
3.1 Introduction
Advances in biological sciences have been critically dependent upon the imaging
techniques available at the time. It follows that understanding the mechanistic
underpinnings of biology in the 21st century relies upon techniques capable of
visualizing the components of entire biological processes in situ, and the emergent
properties of their interactions.
A technique capable of providing such holistic biological insights is electron
cryo-tomography (ECT), an electron microscopy approach that images flash-frozen
specimens in three-dimensions, over scales from molecular to cellular, to resolutions capable of discerning individual macromolecular complexes. Because the
specimen is cryo-preserved without stains, tomographic images are derived from
phase-contrast with the biological material itself and therefore reveal the molecular
J. L. Ferreira Á T. R. S. Matthews-Palmer Á M. Beeby (&)
Department of Life Sciences, Imperial College London, SW7 2AZ London, UK
e-mail: mbeeby@imperial.ac.uk
J. L. Ferreira
e-mail: j.ferreira14@imperial.ac.uk
T. R. S. Matthews-Palmer
e-mail: t.matthews-palmer14@imperial.ac.uk
© 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_3
61