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

Khavnekar, Sagar; Vrbovská, Veronika; Zaoralová, Magda; Kelley, Ron; Beck, Florian; Klumpe, Sven; Kotecha, Abhay; Plitzko, Jürgen; Erdmann, Philipp S.

doi:10.1101/2022.06.16.496417

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

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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;

/persons/resource/persons77726

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

/persons/resource/persons249307

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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https://www.biorxiv.org/content/biorxiv/early/2022/06/16/2022.06.16.496417.full.pdf
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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.