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. 2008 Jan;5(1):53-5.
doi: 10.1038/nmeth1139. Epub 2007 Dec 23.

GraFix: sample preparation for single-particle electron cryomicroscopy

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GraFix: sample preparation for single-particle electron cryomicroscopy

Berthold Kastner et al. Nat Methods. 2008 Jan.

Abstract

We developed a method, named GraFix, that considerably improves sample quality for structure determination by single-particle electron cryomicroscopy (cryo-EM). GraFix uses a glycerol gradient centrifugation step in which the complexes are centrifuged into an increasing concentration of a chemical fixation reagent to prevent aggregation and to stabilize individual macromolecules. The method can be used to prepare samples for negative-stain, cryo-negative-stain and, particularly, unstained cryo-EM.

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