• Medientyp: E-Artikel
  • Titel: A modular platform for automated cryo-FIB workflows
  • Beteiligte: Klumpe, Sven; Fung, Herman KH; Goetz, Sara K; Zagoriy, Ievgeniia; Hampoelz, Bernhard; Zhang, Xiaojie; Erdmann, Philipp S; Baumbach, Janina; Müller, Christoph W; Beck, Martin; Plitzko, Jürgen M; Mahamid, Julia
  • Erschienen: eLife Sciences Publications, Ltd, 2021
  • Erschienen in: eLife
  • Sprache: Englisch
  • DOI: 10.7554/elife.70506
  • ISSN: 2050-084X
  • Schlagwörter: General Immunology and Microbiology ; General Biochemistry, Genetics and Molecular Biology ; General Medicine ; General Neuroscience
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  • Beschreibung: <jats:p>Lamella micromachining by focused ion beam milling at cryogenic temperature (cryo-FIB) has matured into a preparation method widely used for cellular cryo-electron tomography. Due to the limited ablation rates of low Ga<jats:sup>+</jats:sup> ion beam currents required to maintain the structural integrity of vitreous specimens, common preparation protocols are time-consuming and labor intensive. The improved stability of new-generation cryo-FIB instruments now enables automated operations. Here, we present an open-source software tool, SerialFIB, for creating automated and customizable cryo-FIB preparation protocols. The software encompasses a graphical user interface for easy execution of routine lamellae preparations, a scripting module compatible with available Python packages, and interfaces with three-dimensional correlative light and electron microscopy (CLEM) tools. SerialFIB enables the streamlining of advanced cryo-FIB protocols such as multi-modal imaging, CLEM-guided lamella preparation and in situ lamella lift-out procedures. Our software therefore provides a foundation for further development of advanced cryogenic imaging and sample preparation protocols.</jats:p>
  • Zugangsstatus: Freier Zugang