AW: [Histonet] Benchmark Ultra question - follow up

[Angela Bitting]

Augh! I hate when that happens!

>>> "Gudrun Lang" <gu.lang <@t> gmx.at> 4/13/2011 10:44 AM >>>
Thank you all for your replies. After all I have to admit, that the
cause
was human failure. We found, that the protocols have been changed and
noone
did remember. That was after a stopped run, where HIER war clicked off
for
the repetition. And afterward only short HIER was clicked on again
instead
of mild HIER. – stupid me!



Bye Gudrun



  _____  

Von: Lynn Lee [mailto:lynnlee2010 <@t> live.com] 
Gesendet: Mittwoch, 13. April 2011 04:14
An: gu.lang <@t> gmx.at 
Betreff: FW: [Histonet] Benchmark Ultra question - follow up




One last thought- do you filter the new antibody solution you made 
EVERY
time when refilling the prep kit? If not, there could be some
particulates
in the diluent causing a blockage also.  Hope these suggestions help
or
maybe you have already checked this out.

Lynn Lee





  _____  

From: lynnlee2010 <@t> live.com 
To: gu.lang <@t> gmx.at 
Subject: RE: [Histonet] Benchmark Ultra question - follow up
Date: Tue, 12 Apr 2011 20:10:43 -0600

It's possible the flip lid on the prep kit dispenser top is not
completed
closed and seated properly. When the dispense hammer goes down, the
pressure
may not be the same, every time if this is the case.  I always check
the tip
of the dispenser for air bubbles before putting on the machine for
EVERY one
whether it is a prefilled Ventana dispenser or my prep kit dispenser.
Finally something could be clogged in a line somewhere. 

I worked in R & D for Ventana for almost 4 years and have since used
the XT
and Ultra in other labs and never had a problem with incorrect amount
of
reagent being dispensed. I guess there's a first time for anything
though. I
would call Ventana Customer Support. They are great!

Lynn Lee B.S., HT(ASCP)
Tucson, AZ


> From: gu.lang <@t> gmx.at 
> To: histonet <@t> lists.utsouthwestern.edu 
> Date: Tue, 12 Apr 2011 18:23:27 +0200
> Subject: [Histonet] Benchmark Ultra question - follow up
> 
> Today I made a further experiment. I did two titration runs. On one
slide
I
> added the working solution by pipette on the other slide I added it
by
> pushing the filled PrepKit.
> Both stainings came out beautiful. - There has to be something wrong
with
> the dispension-amount. Ventana is informed, I hope they find the
culprit.
> 
> Bye
> Gudrun
> 
> -----Ursprüngliche Nachricht-----
> Von: histonet-bounces <@t> lists.utsouthwestern.edu 
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von
Gudrun
> Lang
> Gesendet: Montag, 11. April 2011 18:48
> An: 'Angela Bitting'
> Cc: histonet <@t> lists.utsouthwestern.edu 
> Betreff: AW: [Histonet] Benchmark Ultra question
> 
> Yes, that could be true. Some of the PrepKits "go harder" than
others.
> Perhaps the pressure of the hammer is not big enough. Sometimes the
nozzles
> are not filled equally. (We press always reagens in the nozzle when
we
> prepare the run.)
> 
> With the Ventana-System there is an additional dilution, because the
> working-solution is added to a reaction-buffer film under the LCS.
Perhaps
> it makes a difference if you dispense the solution with the
pipett-tip
under
> the LCS or if the drop falls on the surface of the LCS. That could be
a
> matter of the LCS-quality, or buffer-quantity , or . I think I'll
become
> mad!
> 
> Our pathologists have an uncertain unhappiness with the overall
staining
> results, but most of the antibodies work well. Perhaps the majority
isn't
> very sensitiv to small changes in the system and the
quality-difference
> isn't big enough, but some are easier to "kill".
> 
> Regards, Gudrun
> 
> 
> 
> 
> 
> _____ 
> 
> Von: Angela Bitting [mailto:akbitting <@t> geisinger.edu] 
> Gesendet: Montag, 11. April 2011 18:06
> An: gu.lang <@t> gmx.at 
> Betreff: Re: [Histonet] Benchmark Ultra question
> 
> 
> 
> When I was trained to do titration runs on the BenchmarkXT and
Ultras, I
was
> told to titrate 100ul of antibody. I have been suspecting for some
time
now
> that the dispensers DON'T dispense a full 100ul. I haven't taken the
time
to
> prove it, but your question may have motivated me.
> 
> That could explain the weak staining. 
> 
> >>> "Gudrun Lang" <gu.lang <@t> gmx.at> 4/11/2011 11:48 AM >>>
> Hi!
> 
> I have some issues with our new Ultra. Since a few days antibodies,
that
> work usually well, show up very faint. For example the TTF1
(Novocastra).
> 
> I ordered a new bottle, made a titration and found, that the "old"
titer
> 1:50 was again well enough. I filled the old PrepKit - and the result
was
> very weak.
> 
> Then I thought, the PrepKit itself is the culprit. I changed the
PrepKit -
> and again the result was very weak. At that time the working-solution
was
> only three days old.
> 
> 
> 
> Has anyone an explanation, why the titration works well and the
automated
> dispension doesn't?
> 
> 
> 
> Regards
> 
> Gudrun Lang
> 
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