[Histonet] Hellerstrom & Hellman anyone?

[Amos Brooks]

Thanks,
	I did, with 50% nitric acid. I would have thought that if there was a problem 
with the glassware it would have shown up in the silver nitrate overnight. The 
silver solution was clear after incubating. The milky mess didn't happen until 
after washing in 95 ETOH then placing in the developing solution. Just to be 
sure though I'll make sure to clean the glassware longer than last time. I'll 
also run a couple of controls and take one out after an hour to reduce just it 
really is over impregnated.

Thanks again,
Amos

Message: 16
Date: Wed, 1 Dec 2010 20:58:20 -0600
From: "Ingles Claire " <CIngles <@t> uwhealth.org>
Subject: RE: [Histonet] Hellerstrom & Hellman anyone?
To: <histonet <@t> lists.utsouthwestern.edu>
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Did you acid clean your glassware before hand?
Claire

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Amos Brooks
Sent: Wed 12/1/2010 12:07 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Hellerstrom & Hellman anyone?



Hi,
        I just did a Hellerstrom & Hellman alcoholic silver stain for 
pancreatic D-
cells. I followed the method described by Bancroft & Stevens. When the slides
were placed into the reducing solution they instantly precipitated into a
toasty mess, the solution turned milky brown and the slides (even around the
sections) picked up a lot of silver precipitate. If anyone has done this of
could offer suggestions about it, I'd really like to trouble shoot the stain a
bit.
        A bit of background about the stain that I thought I should add. 
According to
the published method I used a 10% silver solution in ETOH with 100ul of 1M
nitric acid. The pH of the solution was 5.0. The reducing solution was
alcoholic formalin with 5% pyrogallic acid.
        I thought the silver solution seemed really concentrated (It hardly 
even
disolved). Incubating such a solution overnight at 37 deg C (as indicated in
the method) seemed a bit much. Has anyone heard of modifications of this
stain? There is no sense reinventing the wheel, but I certainly don't want to
repeat it knowing that it will innevitably do the same thing again unless
there is something glaringly obvious that I did wrong.

Thanks,
Amos