Presented in 2012, the RNAscope is one of the more recent RNA in situ hybridization techniques that allows visualization of multiple target genes. RNAscope uses a probe design strategy that amplifies the signal and suppresses the background, which results in increased sensitivity and specificity, see [1].
The protocol automatically detects and quantifies probes for a wide range of genes: ANGPT2, EGLN3, GLP1R, HIF1A, PECAM1, SLC2A1 and dapB. Each cell is classified as 0, 1+, 2+, 3+, or 4+ based on the number of probes it contains as recommended by ACD.