I have been trying to work on smRNA FISH in breast tissue samples with probes and buffers from Stellaris (Biosearch technology).
Unfortunately, I am not getting any signal in either of the tissues. However, the probe works well in the breast tumor derived cell line(MCF-7). I am using 5micron tissue for both fresh frozen and FFPE. Does anyone have any experience with RNA FISH on breast tissue or any tips or pointers to share.??
Please find the protocols I have been using so far :
https://biosearch-cdn.azureedge.net/assetsv6/bti_stellaris_protocol_ffpe_tissue.pdf
https://biosearchassets.blob.core.windows.net/assets/bti_stellaris_protocol_frozen_tissue.pdf
Thank you
Hello Anchal Nigam,
I have exactly the same problem, following the same protocol. Did you find an answer?
Hi Ariadna, we had no success so far. It's very important to have all the procedures right from fixation of tissues to cutting of sections and processing of FISH in RNAse free condition.
Are you also using breast tissue?
Do you do any tissue clearing methods.
We tried proteases treatment in frozen tissue sections and decrosslinking at 85 degrees water bath for 15 minutes plus proteases . However for our tissues it didn't work, but it has worked for many labs.
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