Re: Two Q's

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From:amos brooks <atbrooks@snet.net>
To:denise M m Long-Woodward <denisew2@juno.com>
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Hi,
    It seems logical that the problem #1 is due to poor processing. They may
not have held the tissues in a solution (100% alcohol probably) long enough
which would have prevented subsequent infiltration of either xylene or
paraffin, that would likely cause the patchy appearance.
    As to question #2, I am haven't used this and hence I am curious as to
what our colleges have to say about it ... sorry I couldn't help more.
Amos Brooks

denise M m Long-Woodward wrote:

> Hi folks, two questions if I may:
>
> First, for a co-worker:  They have noticed faint, patchy staining on some
> of their H&E's.  They get stuff from all over the world. In a slide rack,
> most slides are okay, but some have this very faint, almost faded
> appearance on only some of the slides. Seems independent of the position
> in the rack.  They have tried deparaffinizing for over an hour in fresh
> xylene in case it was imcomplete deparaffinization. They have even tried
> complete hydration before the Mayers Hematoxylin step (ten minutes
> Mayers).   Both the eosin and Hematoxylin have the faded appearance.  Any
> ideas?
>
> Second, I'm doing, with success, IL1alpha on transgenic mouse skins.  One
> researcher has EDTA decalcified mouse jaws with attached teeth and
> gingiva. She expects the gingiva to label but it is not. While we are
> trying different options to correct the problem;  does anyone have any
> experience with IL1alpha on EDTA exposed tissues?  Tissues are formalin
> fixed, paraffin embedded (controls and jaws). Using R&D systems mouse
> IL1alpha antibody (made in goat).
>
> Many thanks for your expertise!
> Denise Long Woodward
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