[Histonet] RE: Histonet Digest, Vol 36, Issue 26

From:"Orr, Rebecca"

HI Dawn,

Let's eliminate some variables here.
Here are some suggestions....one variable at a time.

The staining racks themselves can be warped.  Lay them on the counter
and scrutinize them for any areas that don't look level. Load the racks
with slides and place the racks in the stainer, as if starting a run.
Place the leveling device on each and every slide to see if you have a
problem with the racks.  Some of the Lab Vision models allow for a fine
tuning adjustment in the level of the racks while they are in the
holders in the stainer. There may be little screws on either end of the
rack holder on your stainer...but you might need a field service repair
person to help out on this.  Sometimes you can run into issues with the
leveling once  the sink has been removed (the bottom part) and then put
back into place. Also, Are you switching your slide racks between the

I am not able to supply any validated evidence to the following, but in
my experience only, I have found that some paraffins don't give the best
result  when using the one step method in a pressure cooker...go back to
the xylene  to depar for awhile...see if that might be causing some of
the patchy staining.

You might be fighting a few problems all at the same time. 

When you start your run, are you loading the slides out of a buffer
solution or water?  Use a buffer solution...with the tween....I use
Biocare's Automatic wash buffer solution...it works great...Are you
adding tween 20 to your rinse buffer?  Or does is come already in the
rinse buffer? 

I use two drop zones, 100 microliters each...one at the top of the slide
and the other at the bottom...it gets the center covered very nicely...I
am speaking for my lab...
I know you might not have all day to do this, but if it was me having
these problems, I would stand there and watch each and every step occur
on each and every slide...run as full a run as you dare so you can
detect any trends...since some slides work and others don't...  I'll bet
you'll be able to see if you are having any problems with the slides or
the buffers giving you any leveling problems or any reagent spread
problems if you can observe the entire process.
I have some experience with the stainer being level and the racks being
level, but the racks IN the stainer are not...
Please feel free to give me a call, in case you need any clarification
from my scream of consciousness.
Hope this helps.
Assistant Manager, Anatomic Pathology
Evanston Northwestern Healthcare

Message: 7
Date: Mon, 20 Nov 2006 10:42:37 -0500
From: "Olszewski, Dawn" 
Subject: [Histonet] labvision ihc stainer....problems
To: histonet@lists.utsouthwestern.edu
Content-Type: text/plain; charset=utf-8

Dear histonetters,
I have been having IHC issues sporatically for awhile now and would
appreciate any suggestions that might keep me from pulling out my hair.
am only giving out the name of the products we use to give you the
picture.  I am not trying to bash any particular company.  I just need
I have used Dako and Ventanna stainers in the past and know that every
stainer has some issues.  I am now using 2 Labvision stainers and was
wondering if I am the only one having problems.  I have experienced
run failures as well as  only 1-2 antibodies per run not working with no
obvious reason for failure.  
Sometimes the control will be negative and the patient will work or
will work or the control will be positive but the patient will be
(that should have been pos.) so, you never really know if the results
acurate.  But to complicate matters or frustrate me .... the whole
is sporatic with no rhyme or reason and it has happened on BOTH
I can repeat the stains and all work fine or do 3 different slides with
controls and patient side by side and 1 work and 2 fail .  Crazy stuff
that.  I do one step depar and AR in the pressure cooker and also use
with tween 20.  I have had the service reps out here several times
drop zones, etc. because the problem looks to be a flow issue.  I am
ready to  give up.....Maybe I am overlooking something obvious I have
forgotten about .   
Thanks in advance for any suggestions. 
Dawn Olszewski HTL (ASCP) QIHC
South Georgia Medical Center
Valdosta, GA.  

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