Abstract
Fluorescence in situ hybridization (FISH) reveals the abundance and positioning of nucleic acid sequences in fixed samples. Despite recent advances in multiplexed amplification of FISH signals, it remains challenging to achieve high levels of simultaneous amplification and sequential detection with high sampling efficiency and simple workflows. Here, we introduce signal amplification by exchange reaction (SABER), which endows oligo-based FISH probes with long, single-stranded DNA concatemers that aggregate a multitude of short complementary fluorescent imager strands. We show that SABER amplifies RNA and DNA FISH signals (5 to 450-fold) in fixed cells and tissues, apply 17 orthogonal amplifiers against chromosomal targets simultaneously, and detect mRNAs with high efficiency. We further apply 10-plexSABER-FISH to identify in vivo introduced enhancers with cell type-specific activity in the mouse retina. SABER represents a simple and versatile molecular toolkit for rapid and cost-effective multiplexed imaging of nucleic acid targets.
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