Abstract
Here, we present a strategy to identify microcrystals from initial protein crystallization screen experiments and to optimize diffraction quality of those crystals using negative stain transmission electron microscopy (TEM) as a guiding technique. The use of negative stain TEM allows visualization along the process and thus enables optimization of crystal diffraction by monitoring the lattice quality of crystallization conditions. Nanocrystals bearing perfect lattices are seeded and can be used for MicroED as well as growing larger crystals for X-ray and free electron laser (FEL) data collection.
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Acknowledgments
This work was supported by NIH grant R01GM112686 (G.C.), P50GM082251 (G.C. and S.W.), and BioXFEL-STC1231306 (S.W). S.V. acknowledges support from grant R01GM097082. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of NIGMB or NIH.
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Weiss, S., Vergara, S., Lin, G., Calero, G. (2021). Detection of Microcrystals for CryoEM. 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_14
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DOI: https://doi.org/10.1007/978-1-0716-0966-8_14
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