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Updated: Jan 20, 2026
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Published on: November 20, 2021
Qualitative Analyses of Polishing and Precoating FIB Milled Crystals for MicroED
Michael W Martynowycz1, Wei Zhao2, Johan Hattne1
1Howard Hughes Medical Institute, University of California Los Angeles, Los Angeles, CA, USA; Departments of Biological Chemistry and Physiology, University of California Los Angeles, Los Angeles, CA, USA.
Abstract:
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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