[Histonet] Ventana IHC Staining

Douglas D Deltour doug <@t> ppspath.com
Thu Jan 18 13:17:27 CST 2007

A number of things can cause inconsistent staining. Here are a few. 

Barcode label properties Altered
1) The hydrophobic properties of the labels can cause nonstaining of the
issue if it is placed to close to the label.
2) Placement of the labels on the slides-Crooked labels can cause the
solutions to wick off the edge of the slides.
3) Cutting the Labels edges-If the labels are cut it increases the slides
surface area and can cause light staining.
4) Aging-The glue can get old

Silanization of Glass Slides
1) Making of a charged glass slide. Different Vendors have different
2) Consider factors that compromise the adhesion:
A. "Double dipping of slides". This is where the control is picked up first,
then at a later time the patient is picked up and the slide is wet twice.
B. "Stay-On" used with plus slides.
3) Consider the factors that produce unwanted background.
A. Gelatin in the waterbath.
B. Elmer's Glue in the waterbath.
4) Check the storage conditions, age and lot numbers of the slides.

If these steps don't help then do a Vortex Mix test. 

Douglas D. Deltour HT(ASCP)
Histology Supervisor
Professional Pathology Services, PC
One Science Court
Suite 200
Columbia, SC 29203
Fax (803)254-3262
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Chaussey,
Sent: Thursday, January 18, 2007 1:38 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Ventana IHC Staining

We're noticing some issues with our IHC staining.  It's very random but
we've been seeing instances where the positive control tissue stains
fine but the patient tissue doesn't stain at all.  In one instance, we
re-ran the stain manually & it worked fine.  We generally place patient
tissue on the same slide as the control tissue using the red box control
slides (red box is painted on the back of the slide). We have some ideas
but wanted to check in to see if anyone else using Ventana
Instrumentation (Benchmark XT) has seen that problem as well.  Thanks
for any feedback - it's greatly appreciated.    


Leslie J Chaussey


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