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- Title
Plasma FIB milling for the determination of structures in situ.
- Authors
Berger, Casper; Dumoux, Maud; Glen, Thomas; Yee, Neville B.-y.; Mitchels, John M.; Patáková, Zuzana; Darrow, Michele C.; Naismith, James H.; Grange, Michael
- Abstract
Structural biology studies inside cells and tissues require methods to thin vitrified specimens to electron transparency. Until now, focused ion beams based on gallium have been used. However, ion implantation, changes to surface chemistry and an inability to access high currents limit gallium application. Here, we show that plasma-coupled ion sources can produce cryogenic lamellae of vitrified human cells in a robust and automated manner, with quality sufficient for pseudo-atomic structure determination. Lamellae were produced in a prototype microscope equipped for long cryogenic run times (> 1 week) and with multi-specimen support fully compatible with modern-day transmission electron microscopes. We demonstrate that plasma ion sources can be used for structural biology within cells, determining a structure in situ to 4.9 Å, and characterise the resolution dependence on particle distance from the lamella edge. We describe a workflow upon which different plasmas can be examined to further streamline lamella fabrication. The authors harness plasma focused ion beams for pseudo-atomic structure determination, reporting increased throughput and automation in in situ structural biology to elucidate structure-function relationships inside cells and tissues.
- Subjects
FOCUSED ion beams; INDUCTIVELY coupled plasma mass spectrometry; TRANSMISSION electron microscopes; ION beams; ION sources; ION implantation; PLASMA focus; CRYOGENICS; YANG-Mills theory
- Publication
Nature Communications, 2023, Vol 14, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-023-36372-9