[Histonet] RE: WT1 overstaining on Benchmark Ultra

[HERRINGTON, SHEILA]

I have been experiencing the same random over staining issues with WT1 as well. I use the Benchmark Ultras with Ultraview.  It started around the same time as a new lot number changed a few months back.  I have tried everything, but being so random it is hard to find a solution.


Sheila Herrington, IHC Kelowna General
Grade 3, Immunohistochemistry, Histolopathogy
Kelowna General Hospital



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Clare Thornton
Sent: Tuesday, March 26, 2013 8:57 AM
To: 'Dessoye, Michael J'; Histonet
Subject: [Histonet] RE: WT1 overstaining on Benchmark Ultra

I've seen the exact same thing happen...WT-1 antibody from Cell Marque, on Ventana instrumentation.  Except we use Ultraview detection.  I had 3 WT-1 slides running at the same time.  Two were overstained like you describe, one was fine.  The repeats were fine.  The overstained slides almost look like they had been "cooked".  Interesting!


Clare J. Thornton, HTL(ASCP)QIHC 
Assistant Histology Supervisor
Dahl-Chase Diagnostic Services
417 State Street, Suite 540
Bangor, ME 04401
cthornton <@t> dahlchase.com




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Dessoye, Michael J
Sent: Tuesday, March 26, 2013 11:45 AM
To: Histonet
Subject: [Histonet] WT1 overstaining on Benchmark Ultra

Hello Histonet,
 
I have two strange occurrences that I'm trying to get to the bottom of.
Running WT1 from Cell Marque on a Benchmark Ultra using iView detection and the recommended protocol.  Two specimens (both of which happen to be pleural fluid) severely overstained, even leaving the slide with a brown background stain.  This has only happened twice, and the slides were on different positions on different staining runs.  They used the same antibody and detection kit, however other pleural fluid slides with WT1 are fine.  Other slides that run on the same positions also stain ok.  I am repeating the stains on the same tissues to try to repeat this but in the meantime has anyone experienced this condition or have any advice?
 
The only time I've seen this condition before was due to a clogged vortex mixer, but it affected all the slides and is obviously not the case here.
 
Thanks!
 
Michael J. Dessoye, M.S. | Histology Supervisor | Wilkes-Barre General Hospital | An Affiliate of Commonwealth Health | mjdessoye <@t> commonwealthhealth.net | 575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1432 | Fax: 570-552-1526 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

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