Re: Staining problem details

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From:Ms Louise Taylor <> (by way of histonet)
To:histonet <>
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Hi Pam,
I read the first part of your problem but now have erased it, but I
recall you said something about an alcohol rinse after Hx?
It almost sounds as if this is waht is preventing the eosin
"attaching" to the slide, due to some change in surface tension
My solutions to  the problem:
1. Change ALL dewaxing and rehydrating solutions to fresh , unused
2. Agitate the sections in the staining baths if possible. This might
mean just one or two "jiggles" of the rack sporadically while in the
3. Try to avoid doubling up of slides in the staining rack -
particularly zig zagging.
4. Long shot - try changing the processing solutions in the procssor
Hope something works. This can be a very frustrating problem

Louise Taylor
South Africa

On 12 Nov 98 at 11:56, Pam Plumlee wrote:

> 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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