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Nat Methods. 2019 Jun;16(6):533-544. doi: 10.1038/s41592-019-0404-0. Epub 2019 May 20.

SABER amplifies FISH: enhanced multiplexed imaging of RNA and DNA in cells and tissues.

Author information

1
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.
2
Department of Systems Biology, Harvard Medical School, Boston, MA, USA.
3
Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, MA, USA.
4
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. beliveau@uw.edu.
5
Department of Systems Biology, Harvard Medical School, Boston, MA, USA. beliveau@uw.edu.
6
Department of Genome Sciences, University of Washington, Seattle, WA, USA. beliveau@uw.edu.
7
Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, MA, USA. cepko@genetics.med.harvard.edu.
8
Howard Hughes Medical Institute, Chevy Chase, MD, USA. cepko@genetics.med.harvard.edu.
9
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. py@hms.harvard.edu.
10
Department of Systems Biology, Harvard Medical School, Boston, MA, USA. py@hms.harvard.edu.

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 oligonucleotide-based FISH probes with long, single-stranded DNA concatemers that aggregate a multitude of short complementary fluorescent imager strands. We show that SABER amplified RNA and DNA FISH signals (5- to 450-fold) in fixed cells and tissues. We also applied 17 orthogonal amplifiers against chromosomal targets simultaneously and detected mRNAs with high efficiency. We then used 10-plex SABER-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.

PMID:
31110282
PMCID:
PMC6544483
[Available on 2019-11-20]
DOI:
10.1038/s41592-019-0404-0
[Indexed for MEDLINE]

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