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TitleTomo Live: an on-the-fly reconstruction pipeline to judge data quality for cryo-electron tomography workflows.
Journal, issue, pagesActa Crystallogr D Struct Biol, Vol. 80, Issue Pt 4, Page 247-258, Year 2024
Publish dateApr 1, 2024
AuthorsMaxime Comet / Patricia M Dijkman / Reint Boer Iwema / Tilman Franke / Simonas Masiulis / Ruud Schampers / Oliver Raschdorf / Fanis Grollios / Edward E Pryor / Ieva Drulyte /
PubMed AbstractData acquisition and processing for cryo-electron tomography can be a significant bottleneck for users. To simplify and streamline the cryo-ET workflow, Tomo Live, an on-the-fly solution that ...Data acquisition and processing for cryo-electron tomography can be a significant bottleneck for users. To simplify and streamline the cryo-ET workflow, Tomo Live, an on-the-fly solution that automates the alignment and reconstruction of tilt-series data, enabling real-time data-quality assessment, has been developed. Through the integration of Tomo Live into the data-acquisition workflow for cryo-ET, motion correction is performed directly after each of the acquired tilt angles. Immediately after the tilt-series acquisition has completed, an unattended tilt-series alignment and reconstruction into a 3D volume is performed. The results are displayed in real time in a dedicated remote web platform that runs on the microscope hardware. Through this web platform, users can review the acquired data (aligned stack and 3D volume) and several quality metrics that are obtained during the alignment and reconstruction process. These quality metrics can be used for fast feedback for subsequent acquisitions to save time. Parameters such as Alignment Accuracy, Deleted Tilts and Tilt Axis Correction Angle are visualized as graphs and can be used as filters to export only the best tomograms (raw data, reconstruction and intermediate data) for further processing. Here, the Tomo Live algorithms and workflow are described and representative results on several biological samples are presented. The Tomo Live workflow is accessible to both expert and non-expert users, making it a valuable tool for the continued advancement of structural biology, cell biology and histology.
External linksActa Crystallogr D Struct Biol / PubMed:38512070 / PubMed Central
MethodsEM (tomography)
Structure data

EMDB-19666: Cryo-electron tomogram of apoferritin
Method: EM (tomography)

EMDB-19667: Cryo-electron tomogram of keyhole limpet hemocyanin
Method: EM (tomography)

EMDB-19668: Cryo-electron tomogram of TGEV virions
Method: EM (tomography)

EMDB-19669: Cryo-electron tomogram of Influenza virions
Method: EM (tomography)

EMDB-19670: Cryo-electron tomogram of Magnetospirillum gryphiswaldense
Method: EM (tomography)

EMDB-19671: In situ cryo-electron tomogram of Saccharomyces cerevisiae
Method: EM (tomography)

EMDB-19672: Cryo-electron tomogram of keyhole limpet hemocyanin (stage position)
Method: EM (tomography)

EMDB-19673: Cryo-electron tomogram of keyhole limpet hemocyanin (3 um image shift)
Method: EM (tomography)

EMDB-19674: Cryo-electron tomogram of keyhole limpet hemocyanin (6 um image shift)
Method: EM (tomography)

Source
  • Mus musculus (house mouse)
  • Megathura crenulata (invertebrata)
  • Magnetospirillum gryphiswaldense MSR-1 (magnetotactic)
  • Saccharomyces cerevisiae (brewer's yeast)

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