Re: Staining problem details

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From:Mick Rentsch <> (by way of histonet)
To:histonet <>
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Dear Pam,
do not melt the paraffin when drying your slides if you can help it, you run
the risk of creating a situation where water may be trapped under the molten
paraffin, and you will certainly get patchy staining. If you keep your
drying temp to say 45C, the paraffin will still be permeable enough to allow
trapped water to escape.
Mike Rentsch (Downunder)
-----Original Message-----
From: Pam Plumlee <>
To: '' <>
Date: Friday, 13 November 1998 9:57
Subject: Staining problem details

>Hi: To add more details to the staining problem I'm having...first, the
>hematoxylin staining looks normal..just no pink at all in the cytoplasm.
>I move the rack of slides when they are melting off the paraffin in the
>oven, so when staining there is more than enough solution in the
>staining dishes.  I filter the hematox. often and change and rotate the
>alcohols and xylene.  My routine H&E protocol is just about the same as
>in Carson's Histotechnology book.  I have added a little time to
>deparaffination and use D.I. rinses before putting the slides in the
>Eosin Y Alcoholic because our tap H2O is very acid.  The puzzling part
>of this for me is the staining can be uneven on the same section,
>microscopically the section does not look thick or thin and there does
>not seem to be any paraffin left on the unstained section.  My slides
>are not dried in the microwave.  This may just be a ph problem or
>perhaps a processing problem or just a lab gremlin problem.  thanks for
>the suggestions so far. pam

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