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TitleA modular platform for automated cryo-FIB workflows.
Journal, issue, pagesElife, Vol. 10, Year 2021
Publish dateDec 24, 2021
AuthorsSven Klumpe / Herman Kh Fung / Sara K Goetz / Ievgeniia Zagoriy / Bernhard Hampoelz / Xiaojie Zhang / Philipp S Erdmann / Janina Baumbach / Christoph W Müller / Martin Beck / Jürgen M Plitzko / Julia Mahamid /
PubMed AbstractLamella 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 ...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.
External linksElife / PubMed:34951584 / PubMed Central
MethodsEM (subtomogram averaging) / EM (tomography)
Resolution20.8 - 24.0 Å
Structure data

EMDB-13832:
Subtomogram average of 80S ribosomes from a cryo-FIB-lamella of Sum159 human cell line
Method: EM (subtomogram averaging) / Resolution: 24.0 Å

EMDB-13833:
Cryo-electron tomograms from cryo-FIB-lamellae of Sum159 human cell line
Method: EM (tomography)

EMDB-13834:
Subtomogram average of 80S ribosomes from a cryo-FIB-lamella of Sum159 human cell line prepared after cryo-FIB-SEM volume imaging
Method: EM (subtomogram averaging) / Resolution: 24.0 Å

EMDB-13835:
Subtomogram average of 80S ribosomes a cryo-FIB-lamella of Sum159 human cell line prepared after cryo-FIB-SEM volume imaging
Method: EM (tomography)

EMDB-13836:
Cryo-electron tomogram of a cryo-FIB lamella of a HeLa cell
Method: EM (tomography)

EMDB-13837:
Cryo-electron tomogram of a cryo-FIB lamella of Emiliania huxleyi cells
Method: EM (tomography)

EMDB-13838:
Cryo-electron tomogram of a 3D correlated lipid droplet in a cryo-FIB-milled HeLa cell
Method: EM (tomography)

EMDB-13878:
Cryo-electron tomogram from a cryo-FIB lift-out lamella of Drosophila melanogaster egg chambers
Method: EM (tomography)

EMDB-13879:
Subtomogram average of D. melanogaster Ribosomes from tomograms collected on a cryo-lift-out lamella
Method: EM (subtomogram averaging) / Resolution: 20.8 Å

Source
  • Homo sapiens (human)
  • Emiliania huxleyi (eukaryote)
  • Drosophila melanogaster (fruit fly)

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