Tyramide-conjugated DNA barcodes enable signal amplification for multiparametric CODEX imaging

TitleTyramide-conjugated DNA barcodes enable signal amplification for multiparametric CODEX imaging
Publication TypeJournal Article
Year of Publication2022
AuthorsSimonson PD, Valencia I, Patel SS
JournalCommun Biol
Volume5
Issue1
Pagination627
Date Published2022 Jun 27
ISSN2399-3642
Abstract

Multiparametric imaging allows researchers to measure the expression of many biomarkers simultaneously, allowing detailed characterization of cell microenvironments. One such technique, CODEX, allows fluorescence imaging of >30 proteins in a single tissue section. In the commercial CODEX system, primary antibodies are conjugated to DNA barcodes. This modification can result in antibody dysfunction, and development of a custom antibody panel can be very costly and time consuming as trial and error of modified antibodies proceeds. To address these challenges, we developed novel tyramide-conjugated DNA barcodes that can be used with primary antibodies via peroxidase-conjugated secondary antibodies. This approach results in signal amplification and imaging without the need to conjugate primary antibodies. When combined with commercially available barcode-conjugated primary antibodies, we can very quickly develop working antibody panels. We also present methods to perform antibody staining using a commercially available automated tissue stainer and in situ hybridization imaging on a CODEX platform. Future work will include application of the combined tyramide-based and regular CODEX approach to image specific tumors with their immune cell infiltrates, including biomarkers that are currently difficult to image by regular CODEX.

DOI10.1038/s42003-022-03558-8
Alternate JournalCommun Biol
PubMed ID35754060
Related Faculty: 
Paul Simonson, M.D., Ph.D. Sanjay Patel, M.D., MPH

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