Re: Muddy nuclei

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From:Connie McManus <>
To:David Anderson <>,
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At 05:12 AM 06/08/2000 -0500, David Anderson wrote:
>We called them "ghost 
>nuclei." We traced the problem back to a change in the type of paraffin we 
>were using. We had switched from regular Paraplast to Paraplast Extra 
>because it seemed to section better. 

I just experienced this very same problem.  I formerly used Paraplast Xtra
and switched to Fisher's Tissue prep 2.  We got the ghost nuclei, but also,
the eosin staining was pretty faint.  I was using Gill's Hematoxylin from
VWR Scientific.  This is very old stuff, but it changes color in tap water
just the way it is supposed to... or so I thought.  After much
experiementation and having no success, I was left with the fact that both
the hematoxylin and the eosin were no good.  I prepared a batch of Harris
and some fresh eosin, modified the differentiation solution (I came up with
1 second in 1/2 strength acid alcohol) and Wah Lah... fantastic staining.
In fact, this staining is better than it's ever been.  What is it about the
different paraffins that causes this effect in staining??  It isn't just
the fresher H&E alone.  When i was using the Paraplast Xtra, the Gill's and
old eosin worked just fine. 

And now, Histonetters, for an all expense paid trip BY YOU to beautiful
down town Logan, UT, home of fabulous Aggie Ice Cream ... answer this dull
and unexciting question...

What can I do with ALLLLL this OLD Gill Hematoxylin???  *G*

Connie McManus
Veterinary Diagnostics Lab
Utah State University... (Go Aggies! sheesh, and i'm not even a football fan!)
Logan, Utah

When we switched back, the staining 
>improved, but sectioning wasn't as easy. We compromised by using both, 
>regular Paraplast for processing and Paraplast Extra for embedding and that 
>seemed to satisfy everyone. Someone (again I don't remember who)told me that 
>Paraplast Extra was more sensitive to heat and if the slides were dried at 
>very high temperatures it was more difficult to deparaffinize the sections. 
>As I recall, our drying ovens (Lipshaw) were set about 80C, if not higher, 
>in an effort to save time (money?) because the pathologists wanted their 
>slides yesterday. Lowering the drying temps would have been a more logical 
>solution, but when do we allow logic to interfere with speed?
>Hope that helps.
>David Anderson
>Riyadh Armed Forces Hospital
>Riyadh, Saudi Arabia
>Get Your Private, Free E-mail from MSN Hotmail at

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