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| Title | Parallel cryo electron tomography on in situ lamellae. |
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| Journal, issue, pages | Nat Methods, Vol. 20, Issue 1, Page 131-138, Year 2023 |
| Publish date | Dec 1, 2022 |
 Authors | Fabian Eisenstein / Haruaki Yanagisawa / Hiroka Kashihara / Masahide Kikkawa / Sachiko Tsukita / Radostin Danev /  |
| PubMed Abstract | In situ cryo electron tomography of cryo focused ion beam milled samples has emerged in recent years as a powerful technique for structural studies of macromolecular complexes in their native ...In situ cryo electron tomography of cryo focused ion beam milled samples has emerged in recent years as a powerful technique for structural studies of macromolecular complexes in their native cellular environment. However, the possibilities for recording tomographic tilt series in a high-throughput manner are limited, in part by the lamella-shaped samples. Here we utilize a geometrical sample model and optical image shift to record tens of tilt series in parallel, thereby saving time and gaining access to sample areas conventionally used for tracking specimen movement. The parallel cryo electron tomography (PACE-tomo) method achieves a throughput faster than 5 min per tilt series and allows for the collection of sample areas that were previously unreachable, thus maximizing the amount of data from each lamella. Performance testing with ribosomes in vitro and in situ on state-of-the-art and general-purpose microscopes demonstrated the high throughput and quality of PACE-tomo. |
 External links | Nat Methods / PubMed:36456783 |
| Methods | EM (subtomogram averaging) |
| Resolution | 3.1 - 12.1 Å |
| Structure data |  EMDB-33115: Subtomogram average of 70S ribosome  EMDB-33116: Subtomogram average of 70S ribosome (50S aligned)  EMDB-33117: Subtomogram average of 70S ribosome (30S aligned)  EMDB-33118: In situ subtomogram average of 80S ribosome  EMDB-33833: In situ subtomogram average of f-actin  EMDB-33834: Subtomogram average of 70S ribosome (11x11) |
| Source |
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Escherichia coli (E. coli)
