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The tissues are 2 years old - mice kidneys, spleen and liver. Paraffin
is not the culprit as it it fine for routine surg path specimens.
Initially mice were fixed in B5 fixative 2-4 hrs, then into 10%NBF overnight.
The processor had ZN SO4 formalin in the 1rst position and alcoholic zinc
fixative in position 2. The rest of the processing is traditional 70% alc,
2 95%, 3 100%, 2 xylene, 4 paraffin, however this was a 10 hr program,
and with these smaller tissues, an abbreviated program should have been
<p>Tissue is very dry to cut. Warm soapy water has helped to cut
them. H.I.E.R. accomplished with <i>BioGenex antigen retrieval</i>,
<i>DAKO Target</i> or <i>Biocare Reveal</i> or their <i>Decloaker</i>.
Last 2 solutions done in their pressure cooker. BioGen or DAKO done
in microwave, 800 watts, 6 min at power 4 and sit out for 20 min on the
<p>Charged slides used. I don't know anything about Neoprene.
More info please.
<p>Our evaluation is that it is a processing problem. If that is the case,
how would I get around that. The damage is already done.
<p>Victoria Baker wrote:
<p>How old are your tissues? I'm working with archival
<br>material that wasn't capped and is more than 2 years
<br>old. I've soaked them in the refrigerator overnight
<br>on 2.5% ammonia water soaked paper towels. I work
<br>mainly with rats and mice now, but I've worked with
<br>swine as well. It will not affect IHC protocols.
<p>I do put them on charged slides, but then only dry
<br>them at 60 degrees C for an hour. Have you tried to
<br>coat them with Neoprene? It may be another factor
<br>causing the tissue to come off. Are you doing
<br>retrieval? What is your method? Some are tough on
<br>tissue so it can cause tissue loss. Also some
<br>digestion methods can be rough on tissue.
<p>How were your tissues fixed (NBF, Bouins etc?) What
<br>type of tissue is it?
<p>When you section your tissue, how does it cut?
<br>Shreds, tears rips? It may not be fixation that is
<br>your problem, it may be age of paraffin.
<br>Re-processing won't help at all.
<p>If you can be more specific, I hope I can help.
<p>Vikki Baker, HT (ASCP)
<br>American Health Foundation
<br>Valhalla, New York
<p>--- mbpohl <firstname.lastname@example.org> wrote:
<br>> Any suggestions for keeping dried out - probably
<br>> overly fixed - mice
<br>> tissue on slides for IHC staining? We have tried
<br>> "plus" slides, soaking
<br>> the tissue with warn soapy water, gentle overnight
<br>> 37 degree C
<br>> incubator, 60 degree C overnight incubator,
<br>> microwaving, .....etc.
<br>> Any suggestions welcome.
<br>> Thank you.
<br>> Marylou Pohl
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