[3dem] Lack of Lateral Views
Michael.Radermacher at uvm.edu
Fri Oct 5 16:52:57 PDT 2012
If you are just interested in seeing a few side views I would suggest
thick stain embedding. Uranyl acetate is not a good stain for this.
But for example PTA, Amonium Molybdate, NanoW may work well. UA will
work only if you mix it with Trehalose or similar. I have seen
membrane patches standing on their side in some of these stains.
Important is that the stain must be thick, i.e. embedding. You do this
by drying the grids very fast after wicking the last stain droplet.
Quoting Veer Bhatt <veer82 at gmail.com>:
> Dear all,
> I am working on a multimeric membrane protein that has a strong
> tendency to form rings. The top views of these rings can be
> visualized rather easily using negative-staining EM. However, I am
> unable to find ANY lateral views. Could someone suggest a solution?
> CryoEM is not feasible for me at the moment.
> I use Uranyl Acetate for staining over '300 mesh' carbon (pre)coated
> copper grids.
> Thanks a lot,
> Veer Sandeep Bhatt
Michael Radermacher, Ph.D., Prof.
University of Vermont
Dept. Molecular Physiology and Biophysics
HSRF 120 / 149 Beaumont Ave
Burlington, VT 05405
Tel: + 802 656-4834
Fax: + 802 656-0747
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