A modular platform for automated cryo-FIB workflows
| Autor: | Sara K Goetz, Herman KH Fung, Sven Klumpe, Ievgeniia Zagoriy, Bernhard Hampoelz, Xiaojie Zhang, Philipp S Erdmann, Janina Baumbach, Christoph W Müller, Martin Beck, Jürgen M Plitzko, Julia Mahamid |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Electron Microscope Tomography
QH301-705.5 Science Structural Biology and Molecular Biophysics S. cerevisiae Saccharomyces cerevisiae Emiliania huxleyi General Biochemistry Genetics and Molecular Biology Specimen Handling Animals Humans cryo-lift-out correlative light and electron microscopy Biology (General) cryo-FIB-SEM in situ structural biology automation General Immunology and Microbiology electron microscopy General Neuroscience Cryoelectron Microscopy Haptophyta General Medicine cryo-electron tomography Drosophila melanogaster Medicine Chlamydomonas reinhardtii Software HeLa Cells Research Article Human |
| Zdroj: | eLife eLife, Vol 10 (2021) elife |
| ISSN: | 2050-084X |
| Popis: | 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+ 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|