[Histonet] double immuno staining

[Veronique Andriessen]

Hi folks,

I need some help.

I am struggling with a double immunostaining for somatostatin receptor and
desmin.

All my previous double stainings worked beautifully, but they were performed
on fresh frozen acetone fixed pancreatic tissue.

For staining of somatostatin receptor I have to fix with formaldehyde. I use
20 minutes 0.75% PFA  perfused tissue which is snap frozen embedded in
tissue freezing medium and cut on a cryostat. I was advised to do it this
way to prevent loss of antigenicity.
Morphology is quite good.
This fixation works for the somatostatin receptor staining, but it kills the
desmin signal.
I was able to retrieve most of the desmin signal by EIER for 10 minutes with
trypsin-EDTA, but now the morphology is awful.

Can someone give me some advice?

Sincerely yours,

Veronique Andriessen BAS
Lab. Molecular Liver Cell Biology,
Free University Brussels (VUB)Belgium