[Histonet] Processing mouse seminal vesicles

[Rene J Buesa]

Mouse tissues, any one, will dry out too much with ethanol and xylene.
I advise you to stop using ethanol and xylene and substitute BOTH with iso-propanol at the concentrations and times you are using now.
Instead of the first paraffin, use a mixture 1:1 of iso-propanol and paraffin, followed by the 2 paraffins.
Try it and you will notice the difference (and the saings).
René J.

--- On Wed, 12/10/08, Kathleen Roberts <kgrobert <@t> rci.rutgers.edu> wrote:

From: Kathleen Roberts <kgrobert <@t> rci.rutgers.edu>
Subject: [Histonet] Processing mouse seminal vesicles
To: "'histonet <@t> lists.utsouthwestern.edu'" <histonet <@t> lists.utsouthwestern.edu>
Date: Wednesday, December 10, 2008, 11:54 AM

Does anybody have a protocol for this?  My last batch of these came out VERY dry
and crunchy when run with other tissues on my standard protocol, which is as
follows:
(They are fixed on the benchtop in 10% NBF for 4-5 days, then rinsed out before
processing.)

70%: 30 min
80%: 30 min
95%: 45 min
95%: 45 min
100%: 45 min
100%: 45 min
xylene: 45 min
xylene: 45 min
Paraffin: 30 min
Paraffin: 30min
Paraffin: 30 min

My other thought is that something is up with our VIP 5 processor, though no
error messages are showing up.  Any and all suggestions are most welcome.

Thanks in advance,
Kathleen Roberts
Rutgers University



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