TEM, bio, Ag/Au enhancement

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From:Tamara Howard <howard@cshl.org>
To:Microscopy Listserver <Microscopy@sparc5.microscopy.com>, Histology listserver <histonet@pathology.swmed.edu>
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Greetings!

One of the researchers here is trying to do silver and/or gold enhancement
of an immunolabel in brain slices, but is getting amazing background. We
switched to the gold enhancement because everyone was saying how much
cleaner it is....didn't help. I've seen one report in which the
researchers pre-washed their samples in EDTA, because they felt that they
were getting background from Mg in the tissue that was nucleating silver
deposition - anyone else have any thoughts/experience on this? Anyone else
doing enhancement in tissue slices?

He is using kits to do the enhancement - I'm not sure which companies are
the suppliers, but they aren't supposed to be light-sensitive (my first
culprit). The samples are washed extensively in water before enhancement,
and even the no-antibody controls are "lighting up".

Any suggestions will be greatly appreciated!

Tamara Howard
CSHL




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