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The cryo-EM method microcrystal electron diffraction (MicroED)

Nature Methods volume 16, pages 369–379 (2019)Cite this article

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A Publisher Correction to this article was published on 28 March 2021

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Abstract

In 2013 we established a cryo-electron microscopy (cryo-EM) technique called microcrystal electron diffraction (MicroED). Since that time, data collection and analysis schemes have been fine-tuned, and structures for more than 40 different proteins, oligopeptides and organic molecules have been determined. Here we review the MicroED technique and place it in context with other structure-determination methods. We showcase example structures solved by MicroED and provide practical advice to prospective users.

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Fig. 1: Methods in the field of cryo-EM.
Fig. 2: MicroED overview.
Fig. 3: Improvements in MicroED data quality.
Fig. 4: Examples of novel structures determined by MicroED.
Fig. 5: Crystal identification and sample preparation for MicroED.
Fig. 6: Cryo-FIB milling of a thick crystal.
Fig. 7: Comparison of proteinase K data collected with and without an energy filter.
Fig. 8: Dynamics probed in response to radiation damage.

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Acknowledgements

We thank all of our collaborators and trainees who have contributed either directly or indirectly to the development of MicroED. We thank G. Calero (University of Pittsburgh) for providing Figs. 4c and 5b. The Gonen laboratory is supported by funding from the Howard Hughes Medical Institute (HHMI). The Nannenga laboratory is supported by the US National Institutes of Health (R01GM124152). MicroED was developed at the Janelia Research Campus of HHMI using HHMI funds and Janelia Visitor Program funds.

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  1. Chemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, AZ, USA

    Brent L. Nannenga

  2. Center for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, USA

    Brent L. Nannenga

  3. Howard Hughes Medical Institute, University of California, Los Angeles, Los Angeles, CA, USA

    Tamir Gonen

  4. Departments of Biological Chemistry and Physiology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA

    Tamir Gonen

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Nannenga, B.L., Gonen, T. The cryo-EM method microcrystal electron diffraction (MicroED). Nat Methods 16, 369–379 (2019). https://doi.org/10.1038/s41592-019-0395-x

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