TY - JOUR
T1 - Multiplex digital spatial profiling of proteins and RNA in fixed tissue
AU - Merritt, Christopher R.
AU - Ong, Giang T.
AU - Church, Sarah E.
AU - Barker, Kristi
AU - Danaher, Patrick
AU - Geiss, Gary
AU - Hoang, Margaret
AU - Jung, Jaemyeong
AU - Liang, Yan
AU - McKay-Fleisch, Jill
AU - Nguyen, Karen
AU - Norgaard, Zach
AU - Sorg, Kristina
AU - Sprague, Isaac
AU - Warren, Charles
AU - Warren, Sarah
AU - Webster, Philippa J.
AU - Zhou, Zoey
AU - Zollinger, Daniel R.
AU - Dunaway, Dwayne L.
AU - Mills, Gordon B.
AU - Beechem, Joseph M.
N1 - Publisher Copyright:
© 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2020/5/1
Y1 - 2020/5/1
N2 - Digital Spatial Profiling (DSP) is a method for highly multiplex spatial profiling of proteins or RNAs suitable for use on formalin-fixed, paraffin-embedded (FFPE) samples. The approach relies on (1) multiplexed readout of proteins or RNAs using oligonucleotide tags; (2) oligonucleotide tags attached to affinity reagents (antibodies or RNA probes) through a photocleavable (PC) linker; and (3) photocleaving light projected onto the tissue sample to release PC oligonucleotides in any spatial pattern across a region of interest (ROI) covering 1 to ~5,000 cells. DSP is capable of single-cell sensitivity within an ROI using the antibody readout, with RNA detection feasible down to ~600 individual mRNA transcripts. We show spatial profiling of up to 44 proteins and 96 genes (928 RNA probes) in lymphoid, colorectal tumor and autoimmune tissues by using the nCounter system and 1,412 genes (4,998 RNA probes) by using next-generation sequencing (NGS). DSP may be used to profile not only proteins and RNAs in biobanked samples but also immune markers in patient samples, with potential prognostic and predictive potential for clinical decision-making.
AB - Digital Spatial Profiling (DSP) is a method for highly multiplex spatial profiling of proteins or RNAs suitable for use on formalin-fixed, paraffin-embedded (FFPE) samples. The approach relies on (1) multiplexed readout of proteins or RNAs using oligonucleotide tags; (2) oligonucleotide tags attached to affinity reagents (antibodies or RNA probes) through a photocleavable (PC) linker; and (3) photocleaving light projected onto the tissue sample to release PC oligonucleotides in any spatial pattern across a region of interest (ROI) covering 1 to ~5,000 cells. DSP is capable of single-cell sensitivity within an ROI using the antibody readout, with RNA detection feasible down to ~600 individual mRNA transcripts. We show spatial profiling of up to 44 proteins and 96 genes (928 RNA probes) in lymphoid, colorectal tumor and autoimmune tissues by using the nCounter system and 1,412 genes (4,998 RNA probes) by using next-generation sequencing (NGS). DSP may be used to profile not only proteins and RNAs in biobanked samples but also immune markers in patient samples, with potential prognostic and predictive potential for clinical decision-making.
UR - http://www.scopus.com/inward/record.url?scp=85084478124&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85084478124&partnerID=8YFLogxK
U2 - 10.1038/s41587-020-0472-9
DO - 10.1038/s41587-020-0472-9
M3 - Article
C2 - 32393914
AN - SCOPUS:85084478124
SN - 1087-0156
VL - 38
SP - 586
EP - 599
JO - Nature biotechnology
JF - Nature biotechnology
IS - 5
ER -