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Accepted (AM)Open Access (License Unspecified), Open
Conference proceeding
Signal removal methods for highly multiplexed immunofluorescent staining using antibody conjugated oligonucleotides
IMAGING, MANIPULATION, AND ANALYSIS OF BIOMOLECULES, CELLS, AND TISSUES XVII, v 10881, pp 108810X-108810X-7
01 Feb 2019
PMID: 32280155
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Successful cancer treatment continues to elude modern medicine and its arsenal of therapeutic strategies. Therapy resistance is driven by significant tumor heterogeneity, complex interactions between malignant, microenvironmental and immune cells and cross talk between signaling pathways. Advances in molecular characterization technologies such as next generation sequencing have helped unravel this network of interactions and have vastly affected how cancer is diagnosed and treated. However, the translation of complex genomic analyses to pathological diagnosis remains challenging using conventional immunofluorescence (IF) staining, which is typically limited to 2-5 antigens. Numerous strategies to increase distinct antigen detection on a single sample have been investigated, but all have deleterious effects on the tissue limiting the maximum number of biomarkers that can be imaged on a single sample and none can be seamlessly integrated into routine clinical workflows. To facilitate ready integration into clinical histopathology, we have developed a novel cyclic IF (cycIF) technology based on antibody conjugated oligonucleotides (Ab-oligos). In situ hybridization of complementary oligonucleotides (oligos) facilitates biomarker labeling for imaging on any conventional fluorescent microscope. We have validated a variety of oligo configurations and their respective signal removal strategies capable of diminishing fluorescent signal to levels of autofluorescence before subsequent staining cycles. Robust signal removal is performed without the employment of harsh conditions or reagents, maintaining tissue integrity and antigenicity for higher dimensionality immunostaining of a single sample. Our platform Ab-oligo cycIF technology uses conventional fluorophores and microscopes, allowing for dissemination to a broad audience and congruent integration into clinical histopathology workflows.
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Details
- Title
- Signal removal methods for highly multiplexed immunofluorescent staining using antibody conjugated oligonucleotides
- Creators
- Nathan McMahon - Oregon Health & Science UniversityJocelyn Jones - Oregon Health & Science UniversityJennifer Eng - Oregon Health & Science UniversitySunjong Kwon - Oregon Health & Science UniversityYoung-Hwan Chang - Oregon Health & Science UniversityGuillume Thibault - Oregon Hlth & Sci Univ, Dept Biomed Engn, Portland, OR 97201 USAKoei Chin - Oregon Health & Science UniversityMichel Nederlof - Quantitat Imaging Syst, Pittsburgh, PA 15238 USAJoe Gray - Oregon Health & Science UniversitySummer L. Gibbs - Oregon Health & Science University
- Contributors
- D L Farkas (Editor)A Tarnok (Editor)J F Leary (Editor)
- Publication Details
- IMAGING, MANIPULATION, AND ANALYSIS OF BIOMOLECULES, CELLS, AND TISSUES XVII, v 10881, pp 108810X-108810X-7
- Series
- Proceedings of SPIE
- Publisher
- Spie-Int Soc Optical Engineering
- Number of pages
- 7
- Grant note
- R44 CA224994 / NCI NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI) R44CA224994 / NATIONAL CANCER INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI)
- Resource Type
- Conference proceeding
- Language
- English
- Academic Unit
- Pediatrics
- Web of Science ID
- WOS:000473083400018
- Scopus ID
- 2-s2.0-85065417299
- Other Identifier
- 991021838705204721
UN Sustainable Development Goals (SDGs)
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Engineering, Biomedical
- Medical Laboratory Technology
- Optics