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McMahon N, Jones J, Eng J, et al. Signal removal methods for highly multiplexed immunofluorescent staining using antibody conjugated oligonucleotides. Proc SPIE Int Soc Opt Eng. 2019;10881doi: 10.1117/12.2510573.
McMahon, N., Jones, J., Eng, J., Kwon, S., Chang, Y. H., Thibault, G., Chin, K., Nederlof, M., Gray, J., & Gibbs, S. L. (2019). Signal removal methods for highly multiplexed immunofluorescent staining using antibody conjugated oligonucleotides. Proceedings of SPIE--the International Society for Optical Engineering, 10881. https://doi.org/10.1117/12.2510573
McMahon, Nathan, et al. "Signal removal methods for highly multiplexed immunofluorescent staining using antibody conjugated oligonucleotides." Proceedings of SPIE--the International Society for Optical Engineering vol. 10881 (2019). doi: https://doi.org/10.1117/12.2510573
McMahon N, Jones J, Eng J, Kwon S, Chang YH, Thibault G, Chin K, Nederlof M, Gray J, Gibbs SL. Signal removal methods for highly multiplexed immunofluorescent staining using antibody conjugated oligonucleotides. Proc SPIE Int Soc Opt Eng. 2019 Feb;10881. doi: 10.1117/12.2510573. Epub 2019 Mar 04. PMID: 32280155; PMCID: PMC7145771.
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Proc SPIE Int Soc Opt Eng. 2019 Feb;10881. doi: 10.1117/12.2510573. Epub 2019 Mar 04.

Signal removal methods for highly multiplexed immunofluorescent staining using antibody conjugated oligonucleotides.

Proceedings of SPIE--the International Society for Optical Engineering

Nathan McMahon, Jocelyn Jones, Jennifer Eng, Sunjong Kwon, Young-Hwan Chang, Guillume Thibault, Koei Chin, Michel Nederlof, Joe Gray, Summer L Gibbs

Affiliations

  1. Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR 97201.
  2. Computational Biology Program, Oregon Health and Science University, Portland, OR 97201.
  3. OHSU Center for Spatial Systems Biomedicine, Oregon Health and Science University, Portland, OR 97201.
  4. Quantitative Imaging Systems, Pittsburg, PA 15238.
  5. Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97201.

PMID: 32280155 PMCID: PMC7145771 DOI: 10.1117/12.2510573

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).

Keywords: antibody conjugation; biomarker distribution; cyclic immunofluorescence; fluorescence microscopy; oligonucleotide; photocleavable linker; restriction enzyme; tumor heterogeneity

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