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

被引：325

作者：

Saka, Sinem K. ^{[1
,2
]}

Wang, Yu ^{[1
,2
,3
]}

Kishi, Jocelyn Y. ^{[1
,2
]}

Zhu, Allen ^{[1
,2
]}

Zeng, Yitian ^{[1
,3
]}

Xie, Wenxin ^{[1
,3
]}

Kirli, Koray ^{[4
]}

Yapp, Clarence ^{[5
,6
]}

Cicconet, Marcelo ^{[5
]}

Beliveau, Brian J. ^{[1
,2
,10
]}

Lapan, Sylvain W. ^{[3
]}

Yin, Siyuan ^{[1
,3
]}

Lin, Millicent ^{[1
,3
]}

Boyden, Edward S. ^{[7
]}

Kaeser, Pascal S. ^{[8
]}

Pihan, German ^{[9
]}

Church, George M. ^{[1
,3
]}

Yin, Peng ^{[1
,2
]}

机构：

[1] Harvard Univ, Wyss Inst Biol Inspired Engn, Boston, MA 02115 USA

[2] Harvard Med Sch, Dept Syst Biol, Boston, MA 02115 USA

[3] Harvard Med Sch, Dept Genet, Boston, MA 02115 USA

[4] Harvard Med Sch, Dept Biomed Informat, Boston, MA 02115 USA

[5] Harvard Med Sch, Image & Data Anal Core, Boston, MA 02115 USA

[6] Harvard Med Sch, Lab Syst Pharmacol, Boston, MA 02115 USA

[7] MIT, Media Lab, Cambridge, MA 02139 USA

[8] Harvard Med Sch, Dept Neurobiol, Boston, MA 02115 USA

[9] Beth Israel Deaconess Med Ctr, Dept Pathol, 330 Brookline Ave, Boston, MA 02215 USA

[10] Univ Washington, Dept Genome Sci, Seattle, WA 98195 USA

来源：

NATURE BIOTECHNOLOGY | 2019年 / 37卷 / 09期

基金：

美国国家科学基金会; 美国国家卫生研究院;

关键词：

SIGNAL AMPLIFICATION; RNA; HYBRIDIZATION; PLATFORM; QUANTIFICATION; SENSITIVITY; ALGORITHM;

D O I：

10.1038/s41587-019-0207-y

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

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.

引用

页码：1080 / +

页数：15

共 58 条

[1]

Allan C, 2012, NAT METHODS, V9, P245, DOI [10.1038/NMETH.1896, 10.1038/nmeth.1896]

[2] Multiplexed ion beam imaging of human breast tumors [J].

Angelo, Michael ;

Bendall, Sean C. ;

Finck, Rachel ;

Hale, Matthew B. ;

Hitzman, Chuck ;

Borowsky, Alexander D. ;

Levenson, Richard M. ;

Lowe, John B. ;

Liu, Scot D. ;

Zhao, Shuchun ;

Natkunam, Yasodha ;

Nolan, Garry P. .

NATURE MEDICINE, 2014, 20 (04) :436-+

[3] The quantitative proteome of a human cell line [J].

Beck, Martin ;

Schmidt, Alexander ;

Malmstroem, Johan ;

Claassen, Manfred ;

Ori, Alessandro ;

Szymborska, Anna ;

Herzog, Franz ;

Rinner, Oliver ;

Ellenberg, Jan ;

Aebersold, Ruedi .

MOLECULAR SYSTEMS BIOLOGY, 2011, 7

[4] Regional and cellular distribution of neural visinin-like protein immunoreactivities (VILIP-1 and VILIP-3) in human brain [J].

Bernstein, HG ;

Baumann, B ;

Danos, P ;

Diekmann, S ;

Bogerts, B ;

Gundelfinger, ED ;

Braunewell, KH .

JOURNAL OF NEUROCYTOLOGY, 1999, 28 (08) :655-662

[5] THE USE OF CATALYZED REPORTER DEPOSITION AS A MEANS OF SIGNAL AMPLIFICATION IN A VARIETY OF FORMATS [J].

BOBROW, MN ;

LITT, GJ ;

SHAUGHNESSY, KJ ;

MAYER, PC ;

CONLON, J .

JOURNAL OF IMMUNOLOGICAL METHODS, 1992, 150 (1-2) :145-149

[6] CellProfiler: image analysis software for identifying and quantifying cell phenotypes [J].

Carpenter, Anne E. ;

Jones, Thouis Ray ;

Lamprecht, Michael R. ;

Clarke, Colin ;

Kang, In Han ;

Friman, Ola ;

Guertin, David A. ;

Chang, Joo Han ;

Lindquist, Robert A. ;

Moffat, Jason ;

Golland, Polina ;

Sabatini, David M. .

GENOME BIOLOGY, 2006, 7 (10)

[7]

Chang JB, 2017, NAT METHODS, V14, P593, DOI [10.1038/NMETH.4261, 10.1038/nmeth.4261]

[8] Nanoscale imaging of RNA with expansion microscopy [J].

Chen, Fei ;

Wassie, Asmamaw T. ;

Cote, Allison J. ;

Sinha, Anubhav ;

Alon, Shahar ;

Asano, Shoh ;

Daugharthy, Evan R. ;

Chang, Jae-Byum ;

Marblestone, Adam ;

Church, George M. ;

Raj, Arjun ;

Boyden, Edward S. .

NATURE METHODS, 2016, 13 (08) :679-+

[9] Expansion microscopy [J].

Chen, Fei ;

Tillberg, Paul W. ;

Boyden, Edward S. .

SCIENCE, 2015, 347 (6221) :543-548

[10] Mapping 3D genome organization relative to nuclear compartments using TSA-Seq as a cytological ruler [J].

Chen, Yu ;

Zhang, Yang ;

Wang, Yuchuan ;

Zhang, Liguo ;

Brinkman, Eva K. ;

Adams, Stephen A. ;

Goldman, Robert ;

van Steensel, Bas ;

Ma, Jian ;

Belmont, Andrew S. .

JOURNAL OF CELL BIOLOGY, 2018, 217 (11) :4025-4048

← 1 2 3 4 5 6 →