Tyramide-conjugated DNA barcodes enable signal amplification for multiparametric CODEX imaging. Academic Article uri icon

Overview

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.

publication date

  • June 27, 2022

Research

keywords

  • Antibodies
  • DNA Barcoding, Taxonomic

Identity

PubMed Central ID

  • PMC9234042

Scopus Document Identifier

  • 85132987700

Digital Object Identifier (DOI)

  • 10.1016/0022-1759(89)90104-X

PubMed ID

  • 35754060

Additional Document Info

volume

  • 5

issue

  • 1