[Histonet] processing

[James Watson]

Steve,
 
I routinely use 5% glycerin in the absolute alcohols for my mouse tissues.  this has shortened our soaking times and eliminated the drying artifact.  I started using this years ago on whole cross sections of dog hearts that were processed at a 2.5 cm thickness and on frog leg muscle that was very dry.  I am in the process of completeing some comparative studies and hope to get this published soon.  if you want my processing schedules let me know.
 
James Watson
GNF San Diego
jwatson <@t> gnf.org

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Steven Coakley
Sent: Tue 1/31/2006 10:55 AM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] processing 



Good afternoon,
  
  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.
  
  Thanks everyone,
  
  Steve
  

               
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