Can I use ethanol instead of methanol during dehydratation step in ISH (in situ hybridization) ?

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Kazi Islam

I used ethanol for PFA fixation and paraffin embedding. It worked in regular ISH (BM purple substrate based). However, I am not sure it will work for fluorescent in situ hybridization (FISH) if that is your goal.

Camille E. Dumas

Hi Tomasz,

Yes, definitely. I use EtOH routinely on whole mount ISH, its works very well. You might need to adjust slightly the permeabilisation step (slight increase) depedning on your tissue.

I do not manage to make RNAscope in situ technics work with EtOH dehydration though so I still use MeOH for that.

Best,

Camille