biostudies-literature00450Saka SKUnited States Department of Defense | United States Navy | Office of Naval ResearchNIBIB NIH HHSHoward Hughes Medical InstituteNHLBI NIH HHSNIMH NIH HHSWyss Institute’s Molecular Robotics InitiativeHarvard Medical SchoolNHGRI NIH HHSNINDS NIH HHSDamon Runyon Cancer Research FoundationNIGMS NIH HHSU.S. Department of Health & Human Services | National Institutes of HealthNational Science Foundation1080-1090https://www.ebi.ac.uk/biostudies/studies/S-EPMC6728175biostudies-literatureUnknown37(9)Spatial mapping of proteins in tissues is hindered by limitations in multiplexing, sensitivity and throughput. Here we report immunostaining with signal amplification by exchange reaction (Immuno-SABER), which achieves highly multiplexed signal amplification via DNA-barcoded antibodies and orthogonal DNA concatemers generated by primer exchange reaction (PER). SABER offers independently programmable signal amplification without in situ enzymatic reactions, and intrinsic scalability to rapidly amplify and visualize a large number of targets when combined with fast exchange cycles of fluorescent imager strands. We demonstrate 5- to 180-fold signal amplification in diverse samples (cultured cells, cryosections, formalin-fixed paraffin-embedded sections and whole-mount tissues), as well as simultaneous signal amplification for ten different proteins using standard equipment and workflows. We also combined SABER with expansion microscopy to enable rapid, multiplexed super-resolution tissue imaging. Immuno-SABER presents an effective and accessible platform for multiplexed and amplified imaging of proteins with high sensitivity and throughput.Nature biotechnologyImmuno-SABER enables highly multiplexed and amplified protein imaging in tissues.PMC6728175R01NS083898, RM1 HG008525, R01MH1132791R01MH113349R01 MH1132791UG3HL145600/HuBMAP, 1R01EB018659, 1U01MH106011, 1DP1GM133052, 1R01GM124401R01 MH113349R01 NS083898DP1 GM133052N00014-16-1-2410, N00014-16-1-2182, N00014-18-1-2549RM1 HG008525R01 EB018659U01 MH1060115K99EY028215-02P30 NS072030Dean’s InitiativeCCF-1317291R01 GM124401Graduate Research FellowshipUG3 HL145600Yapp CLapan SWXie WKirli KPihan GYin SKaeser PSBeliveau BJZeng YZhu ACicconet MSaka SKBoyden ESKishi JYChurch GMYin PWang YLin M45falseImmuno-SABER enables highly multiplexed and amplified protein imaging in tissues.Spatial mapping of proteins in tissues is hindered by limitations in multiplexing, sensitivity and throughput. Here we report immunostaining with signal amplification by exchange reaction (Immuno-SABER), which achieves highly multiplexed signal amplification via DNA-barcoded antibodies and orthogonal DNA concatemers generated by primer exchange reaction (PER). SABER offers independently programmable signal amplification without in situ enzymatic reactions, and intrinsic scalability to rapidly amplify and visualize a large number of targets when combined with fast exchange cycles of fluorescent imager strands. We demonstrate 5- to 180-fold signal amplification in diverse samples (cultured cells, cryosections, formalin-fixed paraffin-embedded sections and whole-mount tissues), as well as simultaneous signal amplification for ten different proteins using standard equipment and workflows. We also combined SABER with expansion microscopy to enable rapid, multiplexed super-resolution tissue imaging. Immuno-SABER presents an effective and accessible platform for multiplexed and amplified imaging of proteins with high sensitivity and throughput.2019-01-01T00:00:00Z2019 Sep2024-11-20T11:47:27.259Z2020-05-22T10:17:52ZS-EPMC67281753142781910.1038/s41587-019-0207-y