Goh, J.J.L., Chou, N., Seow, W.Y. et al. Highly specific multiplexed RNA imaging in tissues with split-FISH. Nat Methods (2020).
Abstract:
We present split-FISH, a multiplexed fluorescence in situ hybridization method that leverages a split-probe design to achieve enhanced specificity. Split-FISH reduces off-target background fluorescence, decreases false positives and enables accurate RNA profiling in uncleared tissues. We demonstrate the efficacy of split-FISH on various mouse tissues by quantifying the distribution and abundance of 317 genes in single cells and reveal diverse localization patterns for spatial regulation of the transcriptome in complex tissues.
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Funding Info:
National Medical Research Council of Singapore grant OFYIRG15nov017 (K.H.C.), Agency for Science, Technology, and Research grant I1801E0029 (K.H.C.), AXA Post-Doctoral Fellowship (K.H.C.), A*STAR Graduate Academy (N.C.), National Medical Research Council of Singapore grant OFYIRG18nov-0019 (Z.W.Z.), National University of Singapore Presidential Young Professorship start-up fund (Z.W.Z.)