We wish to use the Click-iT EdU Plus Kits to visualise cell proliferation of mouse splenocytes ex vivo.
One method of detecting and visualising EdU incorporation is fluorescence IHC. However, the protocol recommends paraffin-embedding the tissue before staining. Our problem arrises because we would like to multiplex EdU staining with our antibody of interest, which works much better in OCT-cryopreserved tissue than in paraffin-embedded tissue.
Has anyone used the EdU kits and subsequently frozen the tissue for fluorescent IHC?
I have not personally tried to stain samples with incorporated EdU, however, I have EdU after being frozen at -20 C for years. I do not see any reason why freezing a sample with incorporated EdU wouldn't work, as long as you are performing the EdU staining/detection after you pull the frozen samples out, presumably when you are doing your antibody staining (or immediately before if using PE/PE-conjugated Abs)
Good luck!
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