Smith, Jeffery D. (HSC)
Jeffery-Smith <@t> ouhsc.edu
Tue Jan 31 13:17:56 CST 2006
Sounds to me like too much 100% alcohol. Try using 2 80% and only 2 100% this should eliminate the overhardening yet still give you plenty of dehydration. The next suggestion at that temp. with P/V try cutting back the times in 100% and add some time to the lower grade alcohols.
From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Steven Coakley
Sent: Tue 1/31/2006 12:55 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] processing
I'm working with mouse/rat muscle and trying to fine tune my processing schecdule.
My last run of muscle were very dry. I have a copy of the NSH Animal Processing Manual and trying to strike a balance between those listed on page 5-6. I've noticed most of the protocols do not call for heat or P/V. The scedule I currently use is as follows: 1-70% (hold), 1-80%, 2-95%, 3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 paraffins, 45 minutes each at 55C with P/V. I'm fairly new to research and still trying to make the "mental" switch in handling the different tissue.
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