Plasma FIB milling for the determination of structures in situ

Casper Berger, Maud Dumoux, Thomas Glen, Neville B.-y. Yee, John M. Mitchels, Zuzana Patáková, Michele C. Darrow, James H. Naismith and Michael Grange ()
Additional contact information
Casper Berger: Harwell Science & Innovation Campus
Maud Dumoux: Harwell Science & Innovation Campus
Thomas Glen: Harwell Science & Innovation Campus
Neville B.-y. Yee: Harwell Science & Innovation Campus
John M. Mitchels: Thermo Fisher Scientific Brno s.r.o
Zuzana Patáková: Thermo Fisher Scientific Brno s.r.o
Michele C. Darrow: Harwell Science & Innovation Campus
James H. Naismith: Harwell Science & Innovation Campus
Michael Grange: Harwell Science & Innovation Campus

Nature Communications, 2023, vol. 14, issue 1, 1-12

Abstract: 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.

Date: 2023
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DOI: 10.1038/s41467-023-36372-9

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