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Optimizing Cryo-FIB Lamellas for sub-5Å in situ Structural Biology

MPS-Authors
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Khavnekar,  Sagar
Plitzko, Jürgen / Cryo-EM Technology, Max Planck Institute of Biochemistry, Max Planck Society;

Vrbovská,  Veronika
Max Planck Society;

Zaoralová,  Magda
Max Planck Society;

Kelley,  Ron
Max Planck Society;

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Beck,  Florian
Plitzko, Jürgen / Cryo-EM Technology, Max Planck Institute of Biochemistry, Max Planck Society;

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Klumpe,  Sven
Plitzko, Jürgen / Cryo-EM Technology, Max Planck Institute of Biochemistry, Max Planck Society;

Kotecha,  Abhay
Max Planck Society;

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Plitzko,  Jürgen
Department of Microbiology, Max Planck Institute for Marine Microbiology, Max Planck Society;

Erdmann,  Philipp S.
Max Planck Society;

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引用

Khavnekar, S., Vrbovská, V., Zaoralová, M., Kelley, R., Beck, F., Klumpe, S., Kotecha, A., Plitzko, J., & Erdmann, P. S. (2022). Optimizing Cryo-FIB Lamellas for sub-5Å in situ Structural Biology. bioRxiv: the preprint server for biology. doi:10.1101/2022.06.16.496417.


引用: https://hdl.handle.net/21.11116/0000-0010-33B3-A
要旨
We here present a method based on metallic platinum sputtering that can substantially enhance the quality of subtomogram averages from lamellas and thereby reduce the number of particles needed for high-resolution subtomogram averaging. We provide evidence for the physical background of this improvement and demonstrate its usefulness by producing sub-5Å ribosome averages from yeast.Competing Interest StatementV.V., M.Z., R.K. and A.K. are employees of Thermo Fisher Scientific. The other authors declare no competing interests.