ArticleNucleic acids research2024
Multiplexed in situ protein imaging using DNA-barcoded antibodies with extended hybridization chain reactions.
Article in Nucleic acids research, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Single-cell detection and quantification of the microbiota by MicFLY.Cell host & microbe · 2026Article
- Co-option of Lysosomal Machinery for Sponge Biosilicification.bioRxiv : the preprint server for biology · 2026Article
- Optimizing the hybridization chain reaction-fluorescence in situ hybridization (HCR-FISH) protocol forbioRxiv : the preprint server for biology · 2026Article
- The evolving landscape of spatial proteomics technologies in the AI age.Fundamental research · 2026Review
- Applications and development of in situ nucleic acid visualization techniques.Frontiers in bioengineering and biotechnology · 2026Review
- Anterior-posterior patterning in the chaetognath Spadella cephaloptera informs bilaterian nervous system and tail evolution.Communications biology · 2025Article
- Single-cell quantification of the microbiota by flow cytometry: MicFLY.bioRxiv : the preprint server for biology · 2025Article
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- Signal amplification by cyclic extension enables high-sensitivity single-cell mass cytometry.Nature biotechnology · 2025Article
- Deep-tissue transcriptomics and subcellular imaging at high spatial resolution.Science (New York, N.Y.) · 2025Article
- A platform for multimodalbioRxiv : the preprint server for biology · 2025Article
- Precise immunofluorescence canceling for highly multiplexed imaging to capture specific cell states.Nature communications · 2024Article
- Enablers and challenges of spatial omics, a melting pot of technologies.Molecular systems biology · 2023Review
- NEAT-seq: simultaneous profiling of intra-nuclear proteins, chromatin accessibility and gene expression in single cells.Nature methods · 2022Article
- Spatial mapping of protein composition and tissue organization: a primer for multiplexed antibody-based imaging.Nature methods · 2022Review
- Joint single-cell measurements of nuclear proteins and RNA in vivo.Nature methods · 2021Article
Corrections and comments
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Authors and funding
9 authors.
Funding
Abstract
Antibodies have long served as vital tools in biological and clinical laboratories for the specific detection of proteins. Conventional methods employ fluorophore or horseradish peroxidase-conjugated antibodies to detect signals. More recently, DNA-conjugated antibodies have emerged as a promising technology, capitalizing on the programmability and amplification capabilities of DNA to enable highly multiplexed and ultrasensitive protein detection. However, the nonspecific binding of DNA-conjugated antibodies has impeded the widespread adoption of this approach. Here, we present a novel DNA-conjugated antibody staining protocol that addresses these challenges and demonstrates superior performance in suppressing nonspecific signals compared to previously published protocols. We further extend the utility of DNA-conjugated antibodies for signal-amplified in situ protein imaging through the hybridization chain reaction (HCR) and design a novel HCR DNA pair to expand the HCR hairpin pool from the previously published 5 pairs to 13, allowing for flexible hairpin selection and higher multiplexing. Finally, we demonstrate highly multiplexed in situ protein imaging using these techniques in both cultured cells and tissue sections.
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