[Histonet] microtomy technical question: Protocol used for processing

Rene J Buesa rjbuesa <@t> yahoo.com
Fri Nov 16 09:57:14 CST 2007

Cut your dehydrating and clearing times in half. Eliminate one 95% EthOL and add one station with 100 EthOL-clearing 1:2 instead (after the last 100% EthOL).
  René J.
Eva C Andersson <eca9 <@t> georgetown.edu> wrote:
  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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