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PDBsum entry 6n4u
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References listed in PDB file
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Key reference
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Title
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Collection of continuous rotation microed data from ion beam-Milled crystals of any size.
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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,
545.
[DOI no: ]
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PubMed id
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Abstract
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Microcrystal electron diffraction (MicroED) allows for macromolecular structure
solution from nanocrystals. To create crystals of suitable size for MicroED data
collection, sample preparation typically involves sonication or pipetting a
slurry of crystals from a crystallization drop. The resultant crystal fragments
are fragile and the quality of the data that can be obtained from them is
sensitive to subsequent sample preparation for cryoelectron microscopy as
interactions in the water-air interface can damage crystals during blotting.
Here, we demonstrate the use of a focused ion beam to generate lamellae of
macromolecular protein crystals for continuous rotation MicroED that are of
ideal thickness, easy to locate, and require no blotting optimization. In this
manner, crystals of nearly any size may be scooped and milled to desired
dimensions prior to data collection, thus streamlining the methodology for
sample preparation for MicroED.
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