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Figure 1. Summary of the main steps of the RNAscope procedure.
AMP: signal AMPlifier (1, 2, or 3); HRP: HorseRadish peroxidase; HRP-C: HRP channel (1, 2, or 3); TSA: tyramide signal amplification; ZZ: pair of oligonucleotide probes [with target-specific binding sites covering regions of approximately 50 bases and mRNA target regions covering 300–1,000 nucleotides (with a pool of 6–20 ZZ pairs)]. OR + grey arrow indicates, on Day 2, the repeat of steps involved in additional fluorophore coupling using HRP activity in case of probe multiplexing using other channels (two times if two probes, three times if three probes) or interruption of the procedure until mounting of embryos/larvae on Day 3.