RE: [Histonet] Ventana BenchMark IP stainer

From:"Drew Sally A."

We use both the Nexes and Benchmark and we've not really experienced your particular problem on a large scale- but I might wonder if there is a problem w/ reagent slide volume during the depar/CC step. I'm sure you've probably visually checked the priming of those reagents, but maybe a VMS rep could check the volume delivered, or tell you how to.  Do you have the same problem with immunos that don't require any CC?  We use the same control slide set up and all, but we don't routinely put our slides designated for the BMK in the oven before putting them on-I'm not sure it's necessary (but I also don't think that's the problem.

Sally Ann Drew-MT(ASCP)
Univ. of Wisconsin Hospital & Clinics
IHC/ISH Laboratory 
600 Highland Ave. VAH-DM223
Madison, WI 53792-2472

-----Original Message-----
From: Bauer, Karen []
Sent: Thursday, October 09, 2003 1:38 PM
To: Histonet (E-mail)
Subject: [Histonet] Ventana BenchMark IP stainer


One of our Pathologists is unhappy with some of the results from our
BenchMark IP stainer.  Not all, but some tissues fall off and there are
stains where the tissue seems to be "burnt" or washed out.  We realize that
fixation has a lot to do with the way a stain can turn out, and I've told
him this many times.  I'm sure that everyone has had to deal with "Hurry and
get this on the Processor tonight" and then the next morning we are supposed
to make everything cut, stain, and look perfect.  This isn't always the
case.  There will be well fixed tissue that will look and stain the same

We were having this problem with our Ventana Nexes stainer, but after some
micron and heating time adjustments, things started looking good again.  Now
that we've upgraded to the BenchMark, it's like we are starting from scratch
again.  We've had it since July of this year and our Pathologist would
really like our Nexes back.  The thought of changing all of our IP
procedures again does not sound appealing to me.

Right now, this is the way we cut and prepare our slides for IP staining on
the BenchMark:  Cut sections at 4 to 5 microns and place on a positive and
negative control slide. (We use the charged slides with the red control box
on them.)  We let them air dry for 30 minutes and then we place them in a 60
degree C oven for 30 minutes to 1 hour.  We then place them on the BenchMark
for staining.  

The stains that we are having the most troubles with are the ones using the
CC1.  I've checked our buffers and the pH is fine.  We're having difficulty
with our Pathway Her 2 also.  This uses the CC2 and it ends up looking
washed out.  If we keep it at the mild CC2, there's staining but it's light.
Standard CC2 looks like the tissue is "eaten up or burnt out", and Extended
CC2 makes the tissue pretty much fall off.  This only happens to the pt.
tissue.  Our control tissue seems to stain fine. (Pt. tissue and control
tissue are on the same slide.)

I've talked to our Ventana Rep and their technical personnel and no one has
given me any ideas that we haven't tried already.  Can anybody give me some
hints or solutions to these problems?  Has anyone else out there had these


Karen L. Bauer
Histology Department
Luther Hospital

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