9/4/96
using negative stain (2% PTA), but am having problems with background from
the serum proteins obscuring everything else.
I was wondering if anyone had any suggestions as to alternative stains
and/or methods for reducing the protein background without overly diluting
the sample?
Jerry Gagne
Department of Microscopy and Microanalysis
Abbott Laboratories
Gagnegd@PRDAPP01.PRDAP.MSMAIL.ABBOTT.COM
HBsAg is usually found in serum in very high concentrations, therefor
a 1:10 dilution with PBS is usually sufficient to reduce background proteins.
I have always used 1% ammonium molybdate as the negative stain.
Best of Luck,
Ed Calomeni
Medical College of Ohio
Toledo, OH 43699
emlab@opus.mco.edu
Having occasionally experienced problems with PTA, too, I use Uranyl Acetate
2% supplemented with 1% trehalose and get excellent images of HBsAg.
Regards,
Michel
Michel Deschuyteneer
deschuyt@sbbio.be
I refer you and everyone interested by this technique to the papers by J.
Robin Harris who published superb images of negatively stained KLH obtained
with this additive. The most obvious benefit is a smoother appearance of the
stain and improved resolution.
I should point out however that the improvement may not be dramatic in every
situation: in my experience, results may vary and depend probably also on
the material analyzed and/or its processing.
Michel Deschuyteneer
deschuyt@sbbio.be