ジャーナル: Acta Crystallogr D Struct Biol / 年: 2024 タイトル: Tomo Live: an on-the-fly reconstruction pipeline to judge data quality for cryo-electron tomography workflows. 著者: Maxime Comet / Patricia M Dijkman / Reint Boer Iwema / Tilman Franke / Simonas Masiulis / Ruud Schampers / Oliver Raschdorf / Fanis Grollios / Edward E Pryor / Ieva Drulyte / 要旨: 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 ...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.
集束イオンビーム - 装置: OTHER / 集束イオンビーム - イオン: OTHER / 集束イオンビーム - 電圧: 30 / 集束イオンビーム - 電流: 0.5 / 集束イオンビーム - 時間: 2400 / 集束イオンビーム - 温度: 91 K / 集束イオンビーム - Initial thickness: 7000 / 集束イオンビーム - 最終 厚さ: 150 集束イオンビーム - 詳細: Rough milling with 300-1000 pA, polishing with 10-100 pA.. The value given for _em_focused_ion_beam.instrument is TFS Aquilos 2. This is not in a list of allowed ...集束イオンビーム - 詳細: Rough milling with 300-1000 pA, polishing with 10-100 pA.. The value given for _em_focused_ion_beam.instrument is TFS Aquilos 2. This is not in a list of allowed values {'OTHER', 'DB235'} so OTHER is written into the XML file.
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電子顕微鏡法
顕微鏡
TFS KRIOS
特殊光学系
エネルギーフィルター - 名称: TFS Selectris X / エネルギーフィルター - スリット幅: 10 eV
撮影
フィルム・検出器のモデル: FEI FALCON IV (4k x 4k) 実像数: 37 / 平均電子線量: 3.8 e/Å2