[Histonet] Hellerstrom & Hellman anyone?

[Ingles Claire]

Did you acid clean your glassware before hand?
Claire

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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

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