[Histonet] FISH on imprints and isolated nuclei

[Gudrun Lang]

Hi,

I would like to know if anyone had similar experiences. We tried to
establish FISH on imprints and isolated nuclei in our lab. First we used
superfrost-plus-slides like we do with the FISH-paraffin test. After
hybridization we saw an awful background around the cells, really thick
green or red fluorescence (depending on the filter). It looks like the
nuclei were cemented in this stuff. We were clueless because with
Paraffin-FISH there was no such background.

Now we found out, that after heating the slides in the oven (baking slides
at 56 degrees), the background disappears. So it seems, that the
slide-surface is oxidised in the heat and that the hybridization-reagens
can't adhere on it. On the other side the slides, that were only airdried at
roomtemperature, showed the background.

 

Can anyone confirm this?

 

Bye

Gudrun Lang