A Modular Platform for Streamlining Automated Cryo-FIB Workflows
10.1101/2021.05.19.444745
Crossref posted-content
Cold Spring Harbor Laboratory
[] (246)
Abstract

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 ablation rates of low Ga+ ion beam currents required to maintain the structural integrity of vitreous specimens, current 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 3-dimensional correlative light and electron microscopy (CLEM) tools. The software enables the streamlining of advanced cryo-FIB protocols such as multi-modal imaging, CLEM guided 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.

Bibliography

Klumpe, S., Fung, H. K. H., Goetz, S. K., Zagoriy, I., Hampoelz, B., Zhang, X., Erdmann, P. S., Baumbach, J., Müller, C. W., Beck, M., Plitzko, J. M., & Mahamid, J. (2021). A Modular Platform for Streamlining Automated Cryo-FIB Workflows.

Authors 12
  1. Sven Klumpe (first)
  2. Herman K. H. Fung (additional)
  3. Sara K. Goetz (additional)
  4. Ievgeniia Zagoriy (additional)
  5. Bernhard Hampoelz (additional)
  6. Xiaojie Zhang (additional)
  7. Philipp S. Erdmann (additional)
  8. Janina Baumbach (additional)
  9. Christoph W. Müller (additional)
  10. Martin Beck (additional)
  11. Jürgen M. Plitzko (additional)
  12. Julia Mahamid (additional)
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Dates
Type When
Created 4 years, 3 months ago (May 20, 2021, 4:45 a.m.)
Deposited 3 years, 2 months ago (May 25, 2022, 3:47 p.m.)
Indexed 6 months ago (Feb. 20, 2025, 7:30 p.m.)
Issued 4 years, 3 months ago (May 20, 2021)
Published 4 years, 3 months ago (May 20, 2021)
Funders 0

None

@article{Klumpe_2021, title={A Modular Platform for Streamlining Automated Cryo-FIB Workflows}, url={http://dx.doi.org/10.1101/2021.05.19.444745}, DOI={10.1101/2021.05.19.444745}, publisher={Cold Spring Harbor Laboratory}, author={Klumpe, Sven and Fung, Herman K. H. and Goetz, Sara K. and Zagoriy, Ievgeniia and Hampoelz, Bernhard and Zhang, Xiaojie and Erdmann, Philipp S. and Baumbach, Janina and Müller, Christoph W. and Beck, Martin and Plitzko, Jürgen M. and Mahamid, Julia}, year={2021}, month=may }