[Histonet] RE: phobic slides

Walter Benton wbenton <@t> cua.md
Fri Jul 5 12:17:30 CDT 2013


Are you using the slides with the patient and control all in one. If so, is the paint on the top of the slide or the reverse underside? There has been a known problem for years that correlates to what you are describing with the control box slides with paint on the top.

Secondly, have you noticed this phenomenon on one instrument or all? If it is one instrument I would check to make sure the vortex mixers are working properly. This is done easily with a drop of Hematoxylin on the slide and turning the vortex mixers on within the instrument.

Another possibility could be that your instrument may not be lining up with the holes where the slide positions are located. Not knowing which instrument you are using it is hard to say. The benchmark had both slide carousel and dispenser reagent rack moving which often caused misalignment and solutions would not reach the slide, bit instead would be on the drip tray. The XT can have the same problem, but not nearly as often since only the reagents move and not the slides.

As I'm sure you aware slide variability does exist within the pack of slides at times, (I have witnessed this personally with Asian slides specifically, never German or American). It truly depends on the technique being used to charge the slide.

Finally, you may not have seen this with Dako instrumentation since it uses a different mechanism to place reagent on the slide. It uses the drop zones and surfactant to have the solution cover the slide. I would imagine that it is less likely to occur on the Dako unless your slides are truly hydrophobic.

Hope this helps!

Walter Benton HT(ASCP)QIHC
Histology Supervisor
Chesapeake Urology Associates
806 Landmark Drive, Suite 127
Glen Burnie, MD 21061
443-471-5850 (Direct)
410-768-5961 (Lab)
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From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bruce Gapinski [BGapinski <@t> pathgroup.com]
Sent: Friday, July 05, 2013 12:57 PM
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)
Chief Histologist
Marin Medical Laboratories
PathGroup SF


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