ArticleJournal of biomedical optics2020
Oligonucleotide conjugated antibodies permit highly multiplexed immunofluorescence for future use in clinical histopathology.
Article in Journal of biomedical optics, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 34 citations in OpenAlex.
- Cyclic immunofluorescence platform using photocleavable linkers for direct antibody labeling enables cancer phenotyping.bioRxiv : the preprint server for biology · 2026Article
- TRIPODD Enables Single-Cell Quantification of Therapeutic Efficacy.Molecular pharmaceutics · 2026Article
- Circulating Neoplastic-Immune Hybrid Cells Are Biomarkers of Occult Metastasis and Treatment Response in Pancreatic Cancer.Cancers · 2024Article
- Ultraplex microscopy: versatile highly-multiplexed molecular labeling and imaging across scale and resolution.bioRxiv : the preprint server for biology · 2024Article
- Detection of neoplastic-immune hybrid cells with metastatic properties in uveal melanoma.Biomarker research · 2024Article
- A novel dual-release scaffold for fluorescent labels improves cyclic immunofluorescence.RSC chemical biology · 2024Article
- Ultra high content analyses of circulating and tumor associated hybrid cells reveal phenotypic heterogeneity.Scientific reports · 2024Article
- Article
- Detection of neoplastic-immune hybrid cells with metastatic properties in uveal melanoma.Research square · 2023Article
- Multiplex Immunofluorescence: A Powerful Tool in Cancer Immunotherapy.International journal of molecular sciences · 2023Review
- Article
- Extracellular Vesicle Antibody Microarray for Multiplexed Inner and Outer Protein Analysis.ACS sensors · 2022Article
- Circulating Neoplastic-Immune Hybrid Cells Predict Metastatic Progression in Uveal Melanoma.Cancers · 2022Article
- Review
- A framework for multiplex imaging optimization and reproducible analysis.Communications biology · 2022Article
- Oligonucleotide conjugated antibody strategies for cyclic immunostaining.Scientific reports · 2021Article
- TRIPODD: a Novel Fluorescence Imaging Platform for In Situ Quantification of Drug Distribution and Therapeutic Response.Molecular imaging and biology · 2021Article
- Artificial Intelligence and Cellular Segmentation in Tissue Microscopy Images.The American journal of pathology · 2021Review
- Highly multiplexed tissue imaging using repeated oligonucleotide exchange reaction.European journal of immunology · 2021Article
Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
significanceAdvanced genetic characterization has informed cancer heterogeneity and the challenge it poses to effective therapy; however, current methods lack spatial context, which is vital to successful cancer therapy. Conventional immunolabeling, commonplace in the clinic, can provide spatial context to protein expression. However, these techniques are spectrally limited, resulting in inadequate capacity to resolve the heterogenous cell subpopulations within a tumor.
aimWe developed and optimized oligonucleotide conjugated antibodies (Ab-oligo) to facilitate cyclic immunofluorescence (cyCIF), resulting in high-dimensional immunostaining. APPROACH: We employed a site-specific conjugation strategy to label antibodies with unique oligonucleotide sequences, which were hybridized in situ with their complementary oligonucleotide sequence tagged with a conventional fluorophore. Antibody concentration, imaging strand concentration, and configuration as well as signal removal strategies were optimized to generate maximal staining intensity using our Ab-oligo cyCIF strategy.
resultsWe successfully generated 14 Ab-oligo conjugates and validated their antigen specificity, which was maintained in single color staining studies. With the validated antibodies, we generated up to 14-color imaging data sets of human breast cancer tissues.
conclusionsHerein, we demonstrated the utility of Ab-oligo cyCIF as a platform for highly multiplexed imaging, its utility to measure tumor heterogeneity, and its potential for future use in clinical histopathology.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.