Cryo-electron microscopy instrumentation and techniques for life sciences and materials science
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Introduction Cryo-electron microscopy (cryo-EM) is flourishing as a popular method of choice in structural biology and is rapidly developing as the key approach to determining the three-dimensional (3D) structures of large macromolecular assemblies. The great potential of this technology was recognized with the award of the Nobel Prize in Chemistry in 2017 to J. Dubochet, J. Frank, and R. Henderson,1 who pioneered key early advances in the cryo-EM field. The purpose of this article is to review some of the developments in instrumentation and methods that have led to the rise of cryo-EM in the life science community and consider how researchers in the materials community might benefit from these advances. In particular, we will compare transmission electron microscopy (TEM) methods with those associated with scanning transmission electron microscopy (STEM) for cryogenic imaging in both biological science and materials science. In many ways, the development of new detector technologies paved the way for the breakthrough developments in cryo-EM. We briefly discuss these developments in detector technologies, as well as possible future directions in this field. A notable aspect in the development of TEM instrumentation is that through the various revolutions in electron microscopy over much of the last century, which have led to so many exciting advances in our understanding of the structures of
inorganic materials, semiconductors, and biological specimens, the fundamental design of electron optical columns has been largely preserved. This seeming constancy of design, however, has also laid the foundation for achieving extraordinary levels of precision in central elements of modern microscopes such as image and probe correctors, specimen stages, vacuum levels, and automation of specimen delivery. Improvements in the electron microscope have been matched by great advances in technology for ancillary equipment such as image and probe correctors, spectrometers, and detectors, which now allow the recovery of unprecedented levels of information on the 3D structures of proteins and nucleic acids in biological applications (see schematic in Figure 1). The central challenge in biological TEM imaging is radiation damage.2 The revolution in the use of transmission electron imaging in biology has at its core three basic advances, each of which provides a way to partially overcome the effects of damage to organic matter from electron irradiation. First, the development of microscope stages maintained at temperatures close to that of liquid nitrogen provides the means to reduce the damage resulting from irradiation with electron beams. Compared to imaging at room temperature, there is a five- to tenfold reduction in radiation damage at cryogenic temperatures.3 Imaging at cryogenic temperatures allows the generation of images from vitrified biological specimens with
Robert E.A. Williams, Center for Electron Microscopy and Analysis, The Ohio State University, USA; [email protected] David W. McComb, Department of Materials Scien
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