Hello,
I am having difficulties to get smRNA FISH going on fresh frozen breast tissue. The technique worked well on MCF7 cells. We tried the same probes on tissue sample, and we haven't had successful results as yet. We have tried on sections (10 and 5 micron ) with different probes ( including GAPDH). We are getting a lot of background, which makes it more difficult to know if there are any hybridisation or not.
Could anyone please help in troubleshooting or have a protocol that's working well or have tips to reduce the autoflourescence ? We are using Stellaris protocol for fresh frozen tissues.
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