Re: Help!

From:Phillip Huff

I have two possible ideas that you can test, and you can tell me that I am
completely wrong if you want. First, I would check the age of your 
hematoxylin to ensure that it is not degraded. I remember some older posts
on histonet dealing with problems arising from old hematoxylin. I would
make up fresh solutions of hematoxylin on a regular basis after I noticed
any distortion in the nuclear stain. Second, and this may sound a little
funny, I would check that your staining solutions covering your slides are
homogeneous. You may have a higher concentration of hematoxylin staining
part of your sections and the rest may not be staining to the same degree.
If I am completely off base, you may want to better describe what you mean
by "the nuclear detail gradually weakens and fades". Do you mean to say
that the stain intensity is lost, or that the structure of the nucleic
acid itself is being destroyed?

Hope this helps,


Happy Friday everyone. This has been the week from hell, and I need your
> help with a worsening problem. When cutting slides, we place a ribbon on
> each slide. The slides can contain 4 individual sections on it. The
> first section on the slide closest to the frosted (labeled) end is OK,
> but as you look at the subsequent sections, the nuclear detail gradually
> weakens and fades. This happens all on the same slide. It only happens
> with the first biopsy specimens that come off our VIP, and usually does
> not affect any of the larger pieces of tissue. We use reagent alcohol
> and process these biopsies in a VIP separate from the large specimens. I
> reduced the time in the alcohol, and reduced the heat in the retort, but
> I'm having the same problem. As for staining I use a Leica stainer which
> is changed daily, Richard Allen hematoxylin & Eosin with their clarifier
> & bluing solutions.
> Any ideas what could be causing this phenomenon? Any solutions,
> suggestions?
> Noreen
> Noreen Gilman, BS, HT (ASCP) QIHC
> Histopathology Supervisor
> Broward General Medical Center
> Ft. Lauderdale, FL 33316
> 954.355.4358 Phone
> 954.355.4139 Fax
> 954.387.0213 Pager

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