Transmission electron microscopy for the evaluation and optimization of crystal growth
Category
Published on
Type
journal-article
Author
Hilary P. Stevenson and Guowu Lin and Christopher O. Barnes and Ieva Sutkeviciute and Troy Krzysiak and Simon C. Weiss and Shelley Reynolds and Ying Wu and Veeranagu Nagarajan and Alexander M. Makhov and Robert Lawrence and Emily Lamm and Lisa Clark and Timothy J. Gardella and Brenda G. Hogue and Craig M. Ogata and Jinwoo Ahn and Angela M. Gronenborn and James F. Conway and Jean-Pierre Vilardaga and Aina E. Cohen and Guillermo Calero
Citation
Stevenson, H.P. et al., 2016. Transmission electron microscopy for the evaluation and optimization of crystal growth. Acta Crystallographica Section D Structural Biology, 72(5), pp.603–615. Available at: http://dx.doi.org/10.1107/s2059798316001546.
Abstract
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.
