We have uterine tissue samples that we would like to do immunohistochemisty on. Currently the samples are being fixed in RNAlater and frozen at -80. Would we be able to use these types of samples or do they need to be fixed in paraffin or formalin? Thank you.
Hi Laura,
I agree with Sebastian. In order to do IHC the tissue has to be fixed. You can section the tissue to less than 30 microns, mount them on the slide and fix them with ice cold acetone. After which, proceed with ab incubation for your protein of interest. If the tissue samples are stored in freezer longer without cryoprotectant/fixation, chances of cell degradation are high, I would recommend staining them with hematoxylin to verify integrity of the samples.
Parafin and formalin can change the water content of your section i.e. shrink it
I use cryostat samples at 10-15 microns and air dry in a cupboard for a few hours (set up and go for lunch coffee etc..). This stops crystals underneath and the sample sliding off once brought back to room temp. If you store the cryosections (I prefer this method as there's no interference from any other chemical procedures i.e. fixatives) remember to air dry/cupboard dry at least an hour before adding antibodies.
I would defreeze these tissues, fix them in formalin and go for making FFPEs and run standard ihc having epitope retrieval step in the protocol on 4 micron sections in order to get a nice picture of histopathological landscape. Level of resolution would be much better this way.
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