Re: [Histonet] microtomy technical question: Protocol used for processing

Hi Eva

You can cut your times down to around 30 minutes for each station. You are
taking the bound water out of the specimens. You need that water to keep
the specimens from being too dry. That being said animal tissue is dry to
begin with.

There is an excellent Animal Tissue Processing Manual that was published
for NSH and edited by Gayle Callis and Diane Sterchi. This is an excellent
resource. I have used it many times. The manual can be purchased through

If you need further assistance please don't hesitate to give me a call.

Best Regards

Mari Ann Mailhiot BA HT ASCP
Application Specialist
Leica Technical Assistance Center
800 248 0123 x7267
847 236 3063 fax

             Eva C Andersson                                               
             Sent by:           
             histonet-bounces@                                          cc 
                                       [Histonet] microtomy technical      
                                       question: Protocol used for         
             11/16/2007 09:29          processing                          

Hello again,
I recently posted a question regarding hydrating samples before cutting
them. I am having the problem that my samples are very dry and have had to
use glycerol on several of my different tissues.
I have asked the technician who does the processing about the protocol that
is being used as many of you have said that the samples are probably being
overprocessed. They usually use one program but sometimes a slightly
shorter one. Problem has been that in the past some tissues turned up
underprocessed. Most of the tissues we process are mouse and rat.

This is what they use:
(in a Leica ASP300 processor)
70% Ethanol 55min
80% Ethanol 55min
95% Ethanol 55min
95% Ethanol 1hour
100% Ethanol 55min
100% Ethanol 1hour
Clear rite 55min
Clear rite 55min
Clear rite 1hour
Paraffin Wax 1hour
Paraffin Wax 1hour
Paraffin Wax 1hour

Do you have any helpful suggestions? Very tired of having to spend all day
waiting on my samples to hydrate.

Thank in advance,
Eva Permaul
Georgetown University

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