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Structure paper

TitleUsing focus ion beam to prepare crystal lamella for electron diffraction.
Journal, issue, pagesJ Struct Biol, Vol. 205, Issue 3, Page 59-64, Year 2019
Publish dateMar 1, 2019
AuthorsHeng Zhou / Zhipu Luo / Xueming Li /
PubMed AbstractElectron diffraction provides a powerful tool to solve the structures of small protein crystals. However, strong interactions between the electrons and the materials limit the application of the ...Electron diffraction provides a powerful tool to solve the structures of small protein crystals. However, strong interactions between the electrons and the materials limit the application of the electron crystallographic method on large protein crystals with micrometer or larger sizes. Here, we used the focused ion beam (FIB) equipped on the scanning electron microscope (SEM) to mill a large crystal to thin lamella. The influences of the milling on the crystal lamella were observed and investigated, including radiation damage on the crystal surface during the FIB imaging, deformation of the thin crystal lamella, and variation in the diffraction intensities under electron radiation. These observations provide important information to optimize the FIB milling, and hence is important to obtain high-quality crystal samples for routine structure determination of protein crystals using the electron cryo-microscope.
External linksJ Struct Biol / PubMed:30794865
MethodsEM (electron crystallography)
Resolution1.5 - 1.73 Å
Structure data

0864

6lav

EMDB-0864, PDB-6lav:
MicroED structure of lysozyme at 1.73A determained using crystal lamellas prepared by focused ion beam milling
Method: EM (electron crystallography) / Resolution: 1.73 Å

0865

6law

EMDB-0865, PDB-6law:
MicroED structure of proteinase K at 1.50A determained using crystal lamellas prepared by focused ion beam milling
Method: EM (electron crystallography) / Resolution: 1.5 Å

Chemicals

ChemComp-ACT:
ACETATE ION / Acetate

ChemComp-HOH:
WATER / Water

ChemComp-SO4:
SULFATE ION / Sulfate

Source
  • Gallus gallu (chicken)
  • Chicken (chicken)
  • parengyodontium album (fungus)
  • Tritirachium album,Engyodontium album
  • gallus gallus (chicken)
KeywordsHYDROLASE / Lysozyme / MicroED / Focused Ion Beam / crystal lamella / Proteinase K

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