two FISH on one PETS?

yueh1 yueh1 at ms6.hinet.net
Wed Jun 28 20:45:05 EST 2000


In my experience repeat proteolytic digestion and hybridise again only
result exceed background or poor hybridization. Some sample due to improper
fixed procedure can't get good result, we can only prepare duplicate slide
for another try. FISH is a complex procedure, every step will influence the
result, so I think it is not worth to use same slide.

"Fernando Perez" <fernando.perez at uni-koeln.de> ?????
news:3959D7DB.C3DD03EF at uni-koeln.de...
>
>
>
> Hi all,
>
> we analyse archived  formol-fixed, paraffin-embedded tissue sections
(PETs)
> by interphase FISH (procedure after Ensinger et al. 1997, Anticancer Res.
> 17: 4633). In our experience only few samples give satisfying
hybridisation
> results (we used direct labelled alpha sa,tellite probes from Vysis).
Does
> anyone knows if its posible (in case of inappropiate denaturation) to
> repeat proteolytic digestion and hybridise again to rescure poor-signal
> samples?
>
> Thanks for all input
> Fernando
>
>
>
> Dr. Fernando Pérez
> Medizinische Einrichtungen der Uni-Koeln
> Institut fuer Pathologie
> <fernando.perez at uni-koeln.de>
>     Joseph-Stelzmann-Str. 9
>     D- 50931 Koeln
>     Germany
> Work: +49 221 478 63 63
> Fax: +49 221 478 63 60
>







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