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Transmission electron microscopy for the evaluation and optimization of crystal growth
Author(s) -
Stevenson Hilary P.,
Lin Guowu,
Barnes Christopher O.,
Sutkeviciute Ieva,
Krzysiak Troy,
Weiss Simon C.,
Reynolds Shelley,
Wu Ying,
Nagarajan Veeranagu,
Makhov Alexander M.,
Lawrence Robert,
Lamm Emily,
Clark Lisa,
Gardella Timothy J.,
Hogue Brenda G.,
Ogata Craig M.,
Ahn Jinwoo,
Gronenborn Angela M.,
Conway James F.,
Vilardaga Jean-Pierre,
Cohen Aina E.,
Calero Guillermo
Publication year - 2016
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.374
H-Index - 138
ISSN - 2059-7983
DOI - 10.1107/s2059798316001546
Subject(s) - crystallization , protein crystallization , transmission electron microscopy , diffraction , crystallography , materials science , crystal (programming language) , electron diffraction , x ray crystallography , electron crystallography , high resolution transmission electron microscopy , crystal structure , reflection high energy electron diffraction , bragg's law , crystal growth , chemistry , optics , nanotechnology , physics , organic chemistry , computer science , programming language
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.