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

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

Nature biotechnology (2019-08-21)
Sinem K Saka, Yu Wang, Jocelyn Y Kishi, Allen Zhu, Yitian Zeng, Wenxin Xie, Koray Kirli, Clarence Yapp, Marcelo Cicconet, Brian J Beliveau, Sylvain W Lapan, Siyuan Yin, Millicent Lin, Edward S Boyden, Pascal S Kaeser, German Pihan, George M Church, Peng Yin
摘要

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.

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Supelco
三(2-羧乙基)膦 盐酸盐 溶液, 0.5 M, pH 7.0(aqueous solution; pH was adjusted with ammonium hydroxide)
Sigma-Aldrich
丙烯酸钠, 97%
Sigma-Aldrich
马来酰亚胺-PEG2-琥珀酰亚胺酯, ≥95%