[Histonet] RE: phobic slides
Timothy.Morken <@t> ucsfmedctr.org
Fri Jul 5 12:15:04 CDT 2013
I've seen that when using the Dako stainer or manual staining. The small amount of liquid used for the antibody and detection pools up on the slide rather than spreading out. We solved it for those uses with longer soaks in TBS-tween before putting the slides on the stainer and ensuring the rinse buffer and even diluent had tween in it to promote spreading. I wouldn't think that would happen on the Ventana or Bond instruments with their liquid coverslips and coverplates. And you couldn't use the same solution anyway...
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bruce Gapinski
Sent: Friday, July 05, 2013 9:57 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] phobic slides
Has anyone else had incorrect IHC staining due to overly charged slides that are hydrophobic? My Ventana service department was out because I kept complaining about incomplete staining. For instance, the control at the top of the slide stained with both DAB and hematoxylin. But on the very same slide the hematoxylin was missing from the patient tissue.
I told them how this unnerved me. If the same thing happens but the hematoxylin stains and the DAB does not.......I could be looking at a false negative. This may have happened already as far as I know.
The solution according to Ventana is to take 4 slides from each new box of slides (1/2 gross) and do a vortex mix on each of them one at a time. Then you know you have valid slides to stain patient tissues! I have no time for this. And I'm trying to believe it but it's just not scientific. How is it that my results have been free of this "phobia" for over a year? Why didn't I get this (EVER) with Dako instrumentation? The answer, I was told, was that the humidity was a problem. If I were to take this to it's ridiculous conclusion, I would need to inform oncologists not to operate during these incorrectly humidified days. It IS summer so I'm not sure how much humidity is required.
1. What brand of slides do you use?
2. Have you had the "slide phobia" so that it compromised the results? (Slide-phobia is for retired Histologists, no?)
3. (I think I know the answer but I just have to ask) Do you validate slides?
Bruce Gapinsk HT (ASCP)
Marin Medical Laboratories
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