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-Structure paper
タイトル | Serial Lift-Out: sampling the molecular anatomy of whole organisms. |
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ジャーナル・号・ページ | Nat Methods, Year 2023 |
掲載日 | 2023年12月18日 |
著者 | 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 要旨 | 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. |
リンク | Nat Methods / PubMed:38110637 |
手法 | EM (サブトモグラム平均) / EM (トモグラフィー) |
解像度 | 6.9 - 15.0 Å |
構造データ | 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 |
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