Did you acid clean your glassware before hand? Claire ________________________________
From: histonet-boun...@lists.utsouthwestern.edu on behalf of Amos Brooks Sent: Wed 12/1/2010 12:07 PM To: histonet@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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet