AW: [Histonet] Benchmark Ultra question - follow up

Wed Apr 13 09:44:27 CDT 2011

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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