Re: IPX + PAS or H&E

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From:Tony Henwood <> (by way of histonet)
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Dear Lynn,

> Hey Guys, have a question for all of you doing Immunohistochemistry on
>formalin fixed paraffin embedded tissues.
> Has anyone out in histoland counterstained this type of Immuno with PAS
>or Harris Hematoxylin and Eosin?

Yes I have. You can do the routine IPX stain with DAB, followed by
PAT (Periodic Acid Thionin). This should give you the same result as
a PAS.
Or you can use cobalt to intensify and change the colour of
the brown DAB to Blue and follow it with a PAS.

You can use a H&E counterstain, though be sure that the eosin is weak
(eg 1% aqueous eosin - no acid or other red dyes eg phloxine). You
may need to intensify the DAB with (Oh NO) osmium or you could use
amoniacal silver.

Hope this helps,  Tony
Tony Henwood
Senior Scientist
Anatomical Pathology
Royal Prince Alfred Hospital

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