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PDBsum entry 6pkt
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References listed in PDB file
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Key reference
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Title
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Qualitative analyses of polishing and precoating fib milled crystals for microed.
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Authors
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M.W.Martynowycz,
W.Zhao,
J.Hattne,
G.J.Jensen,
T.Gonen.
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Ref.
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Structure, 2019,
27,
1594.
[DOI no: ]
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PubMed id
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Abstract
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Microcrystal electron diffraction (MicroED) leverages the strong interaction
between matter and electrons to determine protein structures from vanishingly
small crystals. This strong interaction limits the thickness of crystals that
can be investigated by MicroED, mainly due to absorption. Recent studies have
demonstrated that focused ion-beam (FIB) milling can thin crystals into
ideal-sized lamellae; however, it is not clear how to best apply FIB milling for
MicroED. Here, the effects of polishing the lamellae, whereby the last few
nanometers are milled away using a low-current gallium beam, are explored in
both the platinum-precoated and uncoated samples. Our results suggest that
precoating samples with a thin layer of platinum followed by polishing the
crystal surfaces prior to data collection consistently led to superior results
as indicated by higher signal-to-noise ratio, higher resolution, and better
refinement statistics. This study lays the foundation for routine and
reproducible methodology for sample preparation in MicroED.
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