Becky wrote:

"1)Under subdued lighting, I remove the slides ..."

And I'm imagining flickering candles and a mysteriious hooded person making
incantations over their latest creation...




Tim Morken


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca
Sent: Monday, August 08, 2005 6:07 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Tween on Ventana Fish


Hi Gudrun,
We are  currently running Her2 Fish on our Ventana with 100% correlation
with other  vendor's chemistries. This is what works very well in my lab:
1)Under subdued lighting, I remove the slides and place them in 2 changes
Reaction buffer. 2)Soak in the second change of reaction buffer for 5
minutes. I believe there is some kind of surfactant in the Reaction buffer
that does the trick. 3)Rinse for a few minutes in running DI water if you
have it...I would only use tap water if you "trust" it! 
4)Soak slides in the 2Xssc for 5 minutes and coverslip out of the SSC with
the Propidium Iodine. As our control sections are on the same slide as the
patient, I use 10 microliters for the control area and 10 microliters on the
patient area and coverslip with 25x50 coverslip. The slides are usually read
the same day and are stored in the freezer until the Docs come to read them.
We also cut an Extra H/E for them to read along with the FISH. Are you set
up with filters for your microscope?  I can help with that, too, if needed.
Good luck Becky

Becky Orr, CLA HT (ASCP)
IHC Lead
Evanston Northwestern Healthcare
ph: 847-570-2771




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