We had similar problems and noticed that our reagents were not "spreading" equally across the slide.
The manufacturer (Biogenex) protocol was to use PBS.
We tried the Dakko TRIS and it covers easily and allows the antibody to spread evenly.
Hope this helps.Sheila Adey HT MLTPort Huron HospitalMichigan> Date: Thu, 17 Apr 2008 17:45:10 -0400> From: Susan.Weber2@va.gov> To: firstname.lastname@example.org; email@example.com> Subject: RE: [Histonet] IHC Staining Variations> CC: > > Trinity-> Are you using the Ventana Benchmark XT equipment? I recently started> using this equipment and in our training class they told us that certain> slides can be hydrophobic and using a Pap Pen can also effect the> staining quality. I switched from Surgipath X slides to Fisher> Superfrost/Plus slides for this very reason. I use the Shandon Double> slide with the rings on them for our FITC stains and did not have a> problem - the ring is on the underside of the slide, not the side you> put the tissue on. Some of the slides which have "boxes" on them also> affect staining quality (unless it is on the underside of the slide of> course). If you are using the Ventana equipment, their Tech Support is> very good and can help you. Even if your not using this equipment, I'd> call the Tech support of your manufacturer and ask for their help.> > > Susan M Weber HT(ASCP)> Histology Supervisor> Louis Stokes Cleveland VA Medical Center> 10701 East Blvd> Cleveland, Ohio 44106> (216) 791-3800 X6154> > -----Original Message-----> From: firstname.lastname@example.org> [mailto:email@example.com] On Behalf Of> firstname.lastname@example.org> Sent: Thursday, April 17, 2008 2:30 PM> To: email@example.com> Subject: [Histonet] IHC Staining Variations> > Hello,> > We have an automated IHC stainer and the doctors are experiencing what> they call, staining variations.? The whole tissue is not staining, just> in parts, when areas of the tissues should be staining.? > > Has anyone experienced this problem and if so, what did you do to remedy> it?? We were staining manually and used a Pap Pen before we switched to> automation and I'm wondering if we are getting areas on the slide that> are not being covered with the reagents.? We have 3 drop zones with> 100ul of reagent dropping in each zone, for a total of 300ul per slide.> > I appreciate any feedback and help.? > > Trinity Whitecomb> Histology Manager> Lab System Works> _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet> > _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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