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| Title | Serial Lift-Out: sampling the molecular anatomy of whole organisms. |
|---|---|
| Journal, issue, pages | Nat Methods, Year 2023 |
| Publish date | Dec 18, 2023 |
 Authors | Oda Helene Schiøtz / Christoph J O Kaiser / Sven Klumpe / Dustin R Morado / Matthias Poege / Jonathan Schneider / Florian Beck / David P Klebl / Christopher Thompson / Jürgen M Plitzko /    |
| PubMed 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. ...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 Å and a subregion of the 11-protofilament microtubule to 13 Å, illustrating how Serial Lift-Out enables the study of multicellular molecular anatomy. |
 External links | Nat Methods / PubMed:38110637 |
| Methods | EM (subtomogram averaging) / EM (tomography) |
| Resolution | 6.9 - 15.0 Å |
| Structure data |  EMDB-17241: C. elegans L1 80S ribosome  EMDB-17242: C. elegans L1 larva 80S ribosome class 1  EMDB-17243: C. elegans L1 larva 80S ribosome class 2  EMDB-17244: C. elegans L1 larva 80S ribosome class 3  EMDB-17245: C. elegans L1 larva 80S ribosome class 4  EMDB-17246: C. elegans L1 larva body wall muscle tomogram  EMDB-17247: C. elegans L1 larva ventral pharyngeal periphery tomogram  EMDB-17248: Tomogram of the nucleus of a C. elegans L1 larva  EMDB-18186: C. elegans L1 larva  EMDB-18187: Focused refinment of 11-protofilament microtubule from C. elegans |
| Source |
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Caenorhabditis elegans (invertebrata)