[3dem] Positive staining with UA

Ken Roux roux at bio.fsu.edu
Thu Mar 3 11:32:48 PST 2011


We have had good luck with uranyl formate. A small amount of ETOH 
(~2%) as a wetting agent helps give a uniform stain distribution and 
we don't find the need to glow discharge. Positive staining usually 
indicates that to much stain was blotted off the grid (or repelled 
from the carbon due to hydrophobicity).


At 02:08 PM 3/3/2011, Terje Dokland wrote:
>Positive staining is very common with uranyl acetate and is also 
>sample specific. Try PTA or ammonium molybdate, which work more like 
>a pure negative stain. Also, make sure to glow discharge the grids 
>before use, as this has a major effect on stain behavior.
>
>On Mar 3, 2011, at 11:54 AM, shashi bhushan wrote:
>
> > Dear All,
> >
> > We are having problems with the negative staining of our 
> complexes with Uranyl Acetate. We are repeatedly having positive 
> staining instead of negative. We have tried many things without any 
> success. Could some one please provide us some useful tips/detailed 
> protocols to avoid the positive staining.
> >
> > Thanks,
> > Regards,
> >
> > Shashi Bhushan
> >
> > <ATT00001..txt>
>
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Kenneth H. Roux, PhD
Kurt G. Hofer Professor of Biological Science
Department of Biological Science
319 Stadium Drive, King Bldg. Rm 3062
PO Box 3064295
Florida State University
Tallahassee FL 32306-4295
Phone   850-644-5037
FAX    850-645-8447

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