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
A modular platform for automated cryo-FIB workflows
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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
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>