Multiplexed in situ hybridization using HCR
Two-stage protocol. Detection stage: probe sets are hybridized to mRNA targets prior to washing unused probes from the sample. Amplification stage: initiators trigger self-assembly of tethered fluorescent amplification polymers prior to washing unused hairpins from the sample.
Experimental timeline. The same two-stage protocol is used independent of the number of target mRNAs.