[3dem] Lack of Lateral Views

[Michael Radermacher]

Dear Veer,

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.

Regards,

Michael

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.
>
> Veer Sandeep Bhatt


-- 
Michael Radermacher, Ph.D., Prof.
University of Vermont
Dept. Molecular Physiology and Biophysics
HSRF 120 / 149 Beaumont Ave
Burlington, VT 05405
USA

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