[Histonet] paraffin carry-over problem

[sgoebel <@t> mirnarx.com]

I had never used clear rite before, but in coming to this lab it is what
they use because they don't want that much xylene around.  However, we
do still have xylene in the lab.  I thought the problem you describe was
just a product of the clear rite so my solution...the first change after
the oven is xylene.  Shh, don't tell on me =)  This has made the
bubbling (lack of staining) and the paraffin in the alcohol go away.  If
you can try this, it seems to work?  Also, I have my oven set to about
75C for 15 or 20 minutes.

Sarah Goebel, BA, HT(ASCP)
Histotechnologist
Mirna Therapeutics
2150 Woodward Street
Suite 100
Austin, Texas  78744
(512)901-0900 ext. 6912




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Patty
Dunlop
Sent: Tuesday, January 11, 2011 3:04 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] paraffin carry-over problem

Hello,

I have a problem with what appears to be flecks of paraffin coming off
of my
slides in my first change of alcohol in my H&E stain.  My pathologist
indicates seeing "drops of unstained areas on the tissue", and I suspect
it
is from unsatisfactory deparaffinization.  I use clear-rite 3 xylene
subst.
and have always done 3 changes at 3 min each before going to 100%
alcohol.
I have contacted the company and they claim that nothing is wrong with
the
product.  I have tried changing out all stations to fresh clear rite 3
and
increasing the time to 5 min each.  I have also increased the temp on my
slide oven (65 for 20min).  All to no avail.  Could this not be the
problem?  Could it be the paraffin?  I use paraplast plus.

I would love to hear ANY suggestions other than what I have already
tried!
I'm starting to really worry now since the problem has persisted for a
few
months.

Thank you!
Patty
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