Abstract
High-resolution image acquisition and structure determination by cryo-electron microscopy is becoming increasingly streamlined. Preparing electron-microscopy grids of suitable quality remains, however, a critical bottleneck. Strategies to achieve particle monodispersity, optimal sample concentration and suitable ice thickness can vary from specimen to specimen. In this book chapter we describe our protocols for negative-stain grid and cryo-grid preparation, which we apply to studying protein–nucleic acid complexes.
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Acknowledgments
We thank the members of our group, past and present, for contributing to the development of the protocols presented here. This work was supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001065), the UK Medical Research Council (FC001065), and the Wellcome Trust (FC001065).
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Goswami, P., Locke, J., Costa, A. (2018). Preparing Frozen-Hydrated Protein–Nucleic Acid Assemblies for High-Resolution Cryo-EM Imaging. In: Lyubchenko, Y. (eds) Nanoscale Imaging. Methods in Molecular Biology, vol 1814. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8591-3_17
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DOI: https://doi.org/10.1007/978-1-4939-8591-3_17
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