Re: [Histonet] IHC Question...

From:"Dana Settembre"

How do you run IHC?   By hand?  Shandon coverplates? automation?
Dana Settembre
University Hospital-UMDNJ
Newark, NJ

>>> "Bauer, Karen"  04/07/06 12:57 PM >>>
Hi to all. 
I recently stained a lymph node case and it has brought up an
interesting question that I can't explain to our Pathologist.  I'm
hoping that someone out there can give me some suggestions.  First of
all, a little background...
The lymph node was processed, by accident, on a "fast run" on our VIP
Tissue Tek.  The processing run was complete in about 3 and a half
hours.  (We thought there were just small biopsies on the run, but the
lymph node cassette was probably thrown in the wrong basket by mistake.)
 Anyway, the lymph node tissue didn't turn out all that bad, but the
fatty tissue did not process adequately (no surprise there!!).  Since
the lymph node tissue cut fairly well, the block was not run back and
re-processed.  H&E's were turned in and stains were ordered.  
Here's my problem:  The CD3 and CD20 stained well and the lymph node
tissue and controls looked as they should, but the LCA on the patient
tissue did not stain at all.  We put the patient tissue on a control
slide, so the control and tissue are stained the same all the way
through the run.  The LCA control looked beautiful, but the lymph node
tissue had no staining.  How can you have positive staining for CD3 and
CD20, but no staining for LCA on a lymph node?  
Since we use a tonsil for our control, we re-ran the LCA with the
tonsil control, a different lymph node case and then the same lymph node
case that we were staining.  Again, the control looked great, but the
lymph node did not stain again.  The different lymph node case that was
also run turned out just fine, so we knew it was tissue specific.  We
thought maybe this lack of staining was due to the insufficient
processing, but then why did the CD3 and CD20 turn out so well? 
Shouldn't they have no staining also?
I'm perplexed and was wondering if anyone else out there has had this
happen to them.
As always, thank you.
Karen Bauer HT(ASCP)
Histology Supervisor
Luther Hospital
Eau Claire, WI
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