[Histonet] FISH baking and dewaxing

[Mark Tarango]

The package insert for PathVysion (HER2 FISH) says to bake overnight at 56
degrees C.  We never do this and our signals are clear, bright, and
punctate.  Many of the slides are just air-dried.  Some are baked for about
20 minutes and I don't notice any difference between them.  I wonder if I
would see a difference with overnight baking...

The aging step we use for cytology preps to be FISHed is 2 minutes in 2x
SCC at 73 degrees C before protease digestion.  I didn't know extended
baking was considered an aging step for tissue sections to be FISHed.
 Thanks Gudrun!

On Fri, Feb 15, 2013 at 7:20 AM, Gudrun Lang <gu.lang <@t> gmx.at> wrote:

> We see better results with FISH-slides baked over night. This is a
> phenomenon called aging, known from cytogenetics.
> It seems, that the time of air-drying and heat leads to a better access of
> the probes to the dna.
> Therefor we let them also in the oven over the weekend.
> Gudrun
>
>
> -----Ursprüngliche Nachricht-----
> Von: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Houston,
> Ronald
> Gesendet: Donnerstag, 14. Februar 2013 20:08
> An: histonet <@t> lists.utsouthwestern.edu
> Betreff: [Histonet] FISH baking and dewaxing
>
> 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
>
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>
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