Abstract
Illuminating a specimen with a parallel electron beam is critical for many experiments in transmission electron microscopy as deviations from this condition cause considerable deterioration of image quality. Carefully establishing parallel illumination is particularly important on two-condenser lens transmission electron microscopes (TEMs) as the parallel illumination condition is limited to a single beam intensity value on these instruments. It was recently shown that a Thermo Fisher Scientific Talos Arctica, a two-condenser lens TEM operating at 200 kV, equipped with a Gatan K2 Summit direct electron detector is capable of resolving frozen-hydrated macromolecules of various sizes and internal symmetries to better than 3 Å resolution using single particle methodologies. A critical aspect of the success of these findings was the careful alignment of the electron microscope to ensure the specimen was illuminated with a parallel electron beam. Here, this chapter describes how to establish parallel illumination conditions in a Talos Arctica TEM for high-resolution cryogenic data collection for structure determination.
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Acknowledgments
I would like thank Gabe Lander, Mengyu Wu, and Mr. Bill Anderson at The Scripps Research Institute (TSRI) and Matthijn Vos for helpful advice and discussion regarding TEM alignments and data acquisition. I would like to thank Mr. Bill Anderson (TSRI), Kevin Corbett (UCSD), Gabe Lander (TSRI), Andres Leschziner (UCSD), Sergey Suslov (UCSD), and Mengyu Wu (TSRI) for critical reading of the chapter and helpful discussions.
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Herzik, M.A. (2021). Setting Up Parallel Illumination on the Talos Arctica for High-Resolution Data Collection. In: Gonen, T., Nannenga, B.L. (eds) cryoEM. Methods in Molecular Biology, vol 2215. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0966-8_6
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DOI: https://doi.org/10.1007/978-1-0716-0966-8_6
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