Transmission electron microscopy for the evaluation and optimization of crystal growth

Hilary P. Stevenson, Guowu Lin, Christopher O. Barnes, Ieva Sutkeviciute, Troy Krzysiak, Simon C. Weiss, Shelley Reynolds, Ying Wu, Veeranagu Nagarajan, Alexander M. Makhov, Robert Lawrence, Emily Lamm, Lisa Clark, Timothy J. Gardella, Brenda Hogue, Craig M. Ogata, Jinwoo Ahn, Angela M. Gronenborn, James F. Conway, Jean Pierre VilardagaAina E. Cohen, Guillermo Calero

Research output: Contribution to journalArticlepeer-review

27 Scopus citations


The crystallization of protein samples remains the most significant challenge in structure determination by X-ray crystallography. Here, the effectiveness of transmission electron microscopy (TEM) analysis to aid in the crystallization of biological macromolecules is demonstrated. It was found that the presence of well ordered lattices with higher order Bragg spots, revealed by Fourier analysis of TEM images, is a good predictor of diffraction-quality crystals. Moreover, the use of TEM allowed (i) comparison of lattice quality among crystals from different conditions in crystallization screens; (ii) the detection of crystal pathologies that could contribute to poor X-ray diffraction, including crystal lattice defects, anisotropic diffraction and crystal contamination by heavy protein aggregates and nanocrystal nuclei; (iii) the qualitative estimation of crystal solvent content to explore the effect of lattice dehydration on diffraction and (iv) the selection of high-quality crystal fragments for microseeding experiments to generate reproducibly larger sized crystals. Applications to X-ray free-electron laser (XFEL) and micro-electron diffraction (microED) experiments are also discussed.

Original languageEnglish (US)
Pages (from-to)603-615
Number of pages13
JournalActa Crystallographica Section D: Structural Biology
Issue number5
StatePublished - 2016


  • Crystal optimization
  • Crystal optimization
  • Micro-electron diffraction
  • Nanocrystallography
  • Structural biology
  • Transmission electron microscopy
  • X-ray free-electron lasers
  • XFELs

ASJC Scopus subject areas

  • Structural Biology


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