Re: [Histonet] Antigen Retrieval w/ B&D Steamer.....

From:Patti Loykasek

Brandon ­ Iıve tried to answer your questions. If you need more info, please
contact me. 

Patti Loykasek
PhenoPath Laboratories
Seattle, WA
> How long do you pre-heat the solutions?
We preheat the solutions for 15 sec- 2 min in the microwave, depending on
the size of container (either  plastic coplin jar or tissue tek container).
Be careful not to superheat your solution & burn yourself!
> How long do you leave in the steamer?
We put the deparaffinized, hydrated sections into 95 degree solution & leave
the slides in the steamer for either 20 or 30 minutes. Each antibody has a
preferred pretreatment at either 20 or 30 minutes. We have several different
solutions we use. Each antibody is titer checked with several different
pretreatments & the  best one becomes our SOP. We start the time when after
adding the slides the pretreatment solution returns to 95 degrees (enlarge
the holes in the steamer lid so that a thermometer will fit) - this brings
some standardization to our pretreatments.
> How long do you cool down?
We cool down for 20 minutes, I have done 15 minutes at other places of
employment. The most important thing is to allow for some slow cool down
that is not immediate & thus not ³flash drying² to your slides.
Hope this helps. 
> Those are a few of the questions that I have so far......If there is any other
> helpful advice using this method it would be greatly appreciated.
> Thanks,
> Brandon Stokes, HT(ASCP)
> Lab Manager
> Cutting Edge Histology Services, LLC
> Tigard, OR
> 503-443-2157
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