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Nat Biotechnol. 2019 Sep;37(9):1080-1090. doi: 10.1038/s41587-019-0207-y. Epub 2019 Aug 19.

Immuno-SABER enables highly multiplexed and amplified protein imaging in tissues.

Saka SK1,2, Wang Y3,4,5, Kishi JY6,7, Zhu A6,7, Zeng Y6,8, Xie W6,8, Kirli K9, Yapp C10,11, Cicconet M10, Beliveau BJ6,7,12, Lapan SW8, Yin S6,8, Lin M6,8, Boyden ES13, Kaeser PS14, Pihan G15, Church GM6,8, Yin P16,17.

Author information

1
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. sinem.saka@wyss.harvard.edu.
2
Department of Systems Biology, Harvard Medical School, Boston, MA, USA. sinem.saka@wyss.harvard.edu.
3
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. yuwang01@fas.harvard.edu.
4
Department of Systems Biology, Harvard Medical School, Boston, MA, USA. yuwang01@fas.harvard.edu.
5
Department of Genetics, Harvard Medical School, Boston, MA, USA. yuwang01@fas.harvard.edu.
6
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.
7
Department of Systems Biology, Harvard Medical School, Boston, MA, USA.
8
Department of Genetics, Harvard Medical School, Boston, MA, USA.
9
Department of Biomedical Informatics, Harvard Medical School, Boston, MA, USA.
10
Image and Data Analysis Core, Harvard Medical School, Boston, MA, USA.
11
Laboratory of Systems Pharmacology, Harvard Medical School, Boston, MA, USA.
12
Department of Genome Sciences, University of Washington, Seattle, WA, USA.
13
Media Lab, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA.
14
Department of Neurobiology, Harvard Medical School, Boston, MA, USA.
15
Pathology Department, Beth Israel Deaconess Medical Center, Boston, MA, USA.
16
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. py@hms.harvard.edu.
17
Department of Systems Biology, Harvard Medical School, Boston, MA, USA. py@hms.harvard.edu.

Abstract

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.

PMID:
31427819
PMCID:
PMC6728175
[Available on 2020-02-19]
DOI:
10.1038/s41587-019-0207-y

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