Re: IHC staining and animal tissue

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From:Victoria Baker <vbaker60@yahoo.com>
To:mbpohl <mbpohl@acsu.buffalo.edu>
Reply-To:
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Hi,

How old are your tissues?  I'm working with archival
material that wasn't capped and is more than 2 years
old.  I've soaked them in the refrigerator overnight
on 2.5% ammonia water soaked paper towels.  I work
mainly with rats and mice now, but I've worked with
swine as well.  It will not affect IHC protocols.

I do put them on charged slides, but then only dry
them at 60 degrees C for an hour. Have you tried to
coat them with Neoprene?  It may be another factor
causing the tissue to come off.  Are you doing
retrieval?  What is your method?  Some are tough on
tissue so it can cause tissue loss.  Also some
digestion methods can be rough on tissue.

How were your tissues fixed (NBF, Bouins etc?) What
type of tissue is it? 

When you section your tissue, how does it cut? 
Shreds, tears rips?  It may not be fixation that is
your problem, it may be age of paraffin. 
Re-processing won't help at all. 

If you can be more specific, I hope I can help.

Vikki Baker, HT (ASCP)
American Health Foundation
Valhalla, New York

phone:  914-789-7127 

--- mbpohl <mbpohl@acsu.buffalo.edu> wrote:
> Any suggestions for keeping dried out - probably
> overly fixed - mice
> tissue on slides for IHC staining?  We have tried
> "plus" slides, soaking
> the tissue with warn soapy water, gentle overnight
> 37 degree C
> incubator, 60 degree C overnight incubator,
> microwaving, .....etc.
> 
> Any suggestions welcome.
> 
> Thank you.
> 
> Marylou Pohl
> mbpohl@acsu.buffalo.edu
> 
> 
> 
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