Preface
The past decade has seen the interest in the field of electron cryomicroscopy (cryo-EM)
greatly expand. This has been driven by new advancements in the design of the transmission
electron cryomicroscope (cryo-TEM) and the development of high-speed direct electron
detectors together with powerful data processing software. These advancements, along with
many others, have brought cryo-EM to the forefront of structural biology. Various modalities of cryo-EM exist including tomography, single particle analysis, 2D electron crystallography, and microcrystal electron diffraction (MicroED) of 3D crystals. This volume
brings together commentary and detailed methods on cutting-edge cryo-EM techniques
from leading practitioners. The volume is organized into four parts each of which focuses on
a particular methodology within the overall field of cryo-EM. While each chapter is placed
within a single method, several chapters will also have valuable insights applicable across all
cryo-EM modalities.
In Part I of this volume, the emphasis is on electron cryotomography, where multiple
images from a sample are collected at different angles and a 3D volume is reconstructed from
these series of images. Tomography is able to provide unprecedented views on the inner
workings of cells and tissues. Chapters within Part I focus on methods for sample preparation, and overviews on how data are collected and processed to produce final 3D reconstructions. Single particle analysis (SPA) is the focus of Part II of this book with several
chapters devoted to this cryo-EM technique. SPA makes use of images of purified biomolecules suspended in vitreous ice to produce high-resolution 3D reconstructions of the
sample. Methods related to single particle sample screening and preparation, microscope
hardware, and data processing and refinement, among others, are found in this chapter.
Parts III and IV focus on the crystal-based cryo-EM methods of electron crystallography of 2D crystals and microcrystal electron diffraction (MicroED) using 3D crystals,
respectively. The chapters in Part III highlight the methodology for studying membrane
protein 2D crystals and new algorithms for processing the data. In Part IV, chapters on the
recently developed MicroED cryo-EM method for the study of 3D crystals are presented. In
these chapters, procedures for crystal screening, automated data collection, structure solution, as well as a general overview for how to best collect data to minimize the effects of
radiation damage in MicroED, are presented.
The broad scope of this volume on cryo-EM methodology should serve as an excellent
resource to current users of cryo-EM and can serve as the foundation for new comers to this
growing field in structural biology.
Los Angeles, CA, USA
Tamir Gonen
Tempe, AZ, USA
Brent L. Nannenga
v
The past decade has seen the interest in the field of electron cryomicroscopy (cryo-EM)
greatly expand. This has been driven by new advancements in the design of the transmission
electron cryomicroscope (cryo-TEM) and the development of high-speed direct electron
detectors together with powerful data processing software. These advancements, along with
many others, have brought cryo-EM to the forefront of structural biology. Various modalities of cryo-EM exist including tomography, single particle analysis, 2D electron crystallography, and microcrystal electron diffraction (MicroED) of 3D crystals. This volume
brings together commentary and detailed methods on cutting-edge cryo-EM techniques
from leading practitioners. The volume is organized into four parts each of which focuses on
a particular methodology within the overall field of cryo-EM. While each chapter is placed
within a single method, several chapters will also have valuable insights applicable across all
cryo-EM modalities.
In Part I of this volume, the emphasis is on electron cryotomography, where multiple
images from a sample are collected at different angles and a 3D volume is reconstructed from
these series of images. Tomography is able to provide unprecedented views on the inner
workings of cells and tissues. Chapters within Part I focus on methods for sample preparation, and overviews on how data are collected and processed to produce final 3D reconstructions. Single particle analysis (SPA) is the focus of Part II of this book with several
chapters devoted to this cryo-EM technique. SPA makes use of images of purified biomolecules suspended in vitreous ice to produce high-resolution 3D reconstructions of the
sample. Methods related to single particle sample screening and preparation, microscope
hardware, and data processing and refinement, among others, are found in this chapter.
Parts III and IV focus on the crystal-based cryo-EM methods of electron crystallography of 2D crystals and microcrystal electron diffraction (MicroED) using 3D crystals,
respectively. The chapters in Part III highlight the methodology for studying membrane
protein 2D crystals and new algorithms for processing the data. In Part IV, chapters on the
recently developed MicroED cryo-EM method for the study of 3D crystals are presented. In
these chapters, procedures for crystal screening, automated data collection, structure solution, as well as a general overview for how to best collect data to minimize the effects of
radiation damage in MicroED, are presented.
The broad scope of this volume on cryo-EM methodology should serve as an excellent
resource to current users of cryo-EM and can serve as the foundation for new comers to this
growing field in structural biology.
Los Angeles, CA, USA
Tamir Gonen
Tempe, AZ, USA
Brent L. Nannenga
v