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Serial Lift-Out: sampling the molecular anatomy of whole organisms

MPS-Authors
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Schiotz,  Oda Helene
Plitzko, Jürgen / Cryo-EM Technology, Max Planck Institute of Biochemistry, Max Planck Society;

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Kaiser,  Christoph J. O.
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;
IMPRS-ML: Martinsried, Max Planck Institute of Biochemistry, Max Planck Society;

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Morado,  D. R.
Briggs, John / Cell and Virus Structure, Max Planck Institute of Biochemistry, Max Planck Society;

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Poege,  Matthias
Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society;

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Schneider,  Jonathan
Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society;
IMPRS-ML: Martinsried, Max Planck Institute of Biochemistry, 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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Klebl,  David P.
Briggs, John / Cell and Virus Structure, Max Planck Institute of Biochemistry, Max Planck Society;

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Plitzko,  Jürgen M.
Plitzko, Jürgen / Cryo-EM Technology, Max Planck Institute of Biochemistry, Max Planck Society;

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Citation

Schiotz, O. H., Kaiser, C. J. O., Klumpe, S., Morado, D. R., Poege, M., Schneider, J., et al. (2023). Serial Lift-Out: sampling the molecular anatomy of whole organisms. Nature Methods. doi:10.1038/s41592-023-02113-5.


Cite as: https://hdl.handle.net/21.11116/0000-000E-1F1D-1
Abstract
Cryo-focused ion beam milling of frozen-hydrated cells and subsequent cryo-electron tomography (cryo-ET) has enabled the structural elucidation of macromolecular complexes directly inside cells. Application of the technique to multicellular organisms and tissues, however, is still limited by sample preparation. While high-pressure freezing enables the vitrification of thicker samples, it prolongs subsequent preparation due to increased thinning times and the need for extraction procedures. Additionally, thinning removes large portions of the specimen, restricting the imageable volume to the thickness of the final lamella, typically <300 nm. Here we introduce Serial Lift-Out, an enhanced lift-out technique that increases throughput and obtainable contextual information by preparing multiple sections from single transfers. We apply Serial Lift-Out to Caenorhabditis elegans L1 larvae, yielding a cryo-ET dataset sampling the worm's anterior-posterior axis, and resolve its ribosome structure to 7 & Aring; and a subregion of the 11-protofilament microtubule to 13 & Aring;, illustrating how Serial Lift-Out enables the study of multicellular molecular anatomy.