Re: IHC staining and animal tissue

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From:mbpohl <>
To:Victoria Baker <>
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<!doctype html public "-//w3c//dtd html 4.0 transitional//en"> <html> 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 used <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 counter. <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: <blockquote TYPE=CITE>Hi, <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>phone:  914-789-7127 <p>--- mbpohl <> 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>> <br>> Any suggestions welcome. <br>> <br>> Thank you. <br>> <br>> Marylou Pohl <br>> <br>> <br>> <br>> <br>__________________________________________________ <br>Do You Yahoo!? <br>Talk to your friends online with Yahoo! Messenger. <br><a href=""></a></blockquote> </html>
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