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Updated: Dec 18, 2025

Highly-Multiplexed Tissue Imaging with Raman Dyes
Published on: April 21, 2022
Highly specific multiplexed RNA imaging in tissues with split-FISH
Jolene Jie Lin Goh1, Nigel Chou1, Wan Yi Seow1
1Genome Institute of Singapore, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore.
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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