[Histonet] Benchmark Ultra question - follow up

[Gudrun Lang]

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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