[Histonet] FISH baking and dewaxing

[Mark Tarango]

Hi Ronnie,

FISH pretreatment can be harsh and that might explain the long baking
(don't want the tissue to fall off).  On the other hand, I've put a slide
in the oven for 5 minutes and had the tissue stay on in a rush.

I think the dewaxing can be shortened without affecting the FISH signals.
 Our molecular director has us validate any minor change to the
pretreatment protocol, so depending on the director's opinion it could mean
a re-validation.

Mark T.



On Thu, Feb 14, 2013 at 11:08 AM, Houston, Ronald <
Ronald.Houston <@t> nationwidechildrens.org> wrote:

> Can someone please explain why paraffin-embedded slides for FISH need to
> be baked for so long and have extensive dewaxing in xylenes and subsequent
> alcohol pretreatment?
> Our Molecular group bake for >1 hr at 65C, and then treat sections in 3
> changes of xylene 15 min each, and 100% ethanol 15 minutes each x2 before
> air-drying. I have asked them why so long and get the standard response --
> "that's the way we always do it"!
>
> Thanks
> Ronnie
>
> Ronnie Houston, MS HT(ASCP)QIHC
> Anatomic Pathology Manager
> ChildLab, a Division of Nationwide Children's Hospital
> www.childlab.com
>
> 700 Children's Drive
> Columbus, OH 43205
> (P) 614-722-5450
> (F) 614-722-2899
> ronald.houston <@t> nationwidechildrens.org<mailto:
> ronald.houston <@t> nationwidechildrens.org>
> www.NationwideChildrens.org<http://www.nationwidechildrens.org/>
>
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