Dear colleagues,
Recently, Thomas Martin and Anthony Fitzpatrick in my group, together with Andreas Boland from the Barford group, have successfully used graphene oxide as a support layer for cryo-EM grids on various projects (e.g. see
http://dx.doi.org/10.1038/ncomms11293). Graphene oxide may be used, much like a thin film of amorphous carbon, to concentrate particles on the grid, to modify orientational distributions, or to convince
particles to go over the holes. However, the background signal from a single layer of graphene oxide is significantly less than for a thin film of amorphous carbon, making it better suited for small complexes.
The use of graphene oxide for this purpose was first proposed by Rado Pantelic et al. in the Plitzko group (see
http://dx.doi.org/10.1016/j.jsb.2009.12.020). Thomas, Andreas and Anthony made minor modifications to their procedure, i.e. they introduced washing steps to have a more reproducible coverage
of single graphene oxide layers. In the hope that this procedure is useful for others, Thomas has kindly prepared a video on how to prepare these grids. This procedure takes only a few minutes and is generally perceived as much easier than making and depositing
a thin film of amorphous carbon. The video is available through figshare:
http://dx.doi.org/10.6084/m9.figshare.3178669. Please direct questions about the practicalities of this method directly to Thomas (CC), and don't forget to cite the Pantelic paper when using graphene oxide grids.
Best wishes,
Sjors
--
Sjors Scheres
MRC Laboratory of Molecular Biology
Francis Crick Avenue, Cambridge Biomedical Campus
Cambridge CB2 0QH, U.K.
tel: +44 (0)1223 267061
http://www2.mrc-lmb.cam.ac.uk/groups/scheres
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