[Histonet] microtomy technical question: Protocol used forprocessing

Mike Pence mpence <@t> grhs.net
Fri Nov 16 10:09:59 CST 2007

Give the fact that your tissue most likely contain very little
fibroadipose tissue and that your gross tissue sections are most likely
half of what a human tissue is. I would say that your are over
processing your tissue by at least twice.  Cut your ETOH times in half
to start and get rid of one 95% - cut your clear rite in half and add
another station of clear rite. This is where I wold start.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Eva C
Sent: Friday, November 16, 2007 9:29 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] microtomy technical question: Protocol used

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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