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PICASSO: Ultra-multiplexed fluorescence imaging of biomolecules through single-round imaging and blind source unmixing

Junyoung Seo, Yeonbo Sim, Jeewon Kim, Hyunwoo Kim, In Cho, Young-Gyu Yoon, Jae-Byum Chang
doi: https://doi.org/10.1101/2021.01.27.428247
Junyoung Seo
1Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Korea
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Yeonbo Sim
2Department of Biomedical Engineering, Sungkyunkwan University, Suwon, Korea
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Jeewon Kim
3School of Electrical Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Korea
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Hyunwoo Kim
1Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Korea
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In Cho
1Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Korea
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Young-Gyu Yoon
3School of Electrical Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Korea
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Jae-Byum Chang
1Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Korea
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  • For correspondence: jbchang03@gmail.com
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ABSTRACT

Ultra-multiplexed fluorescence imaging of biomolecules is essential to studying heterogeneous biological systems. However, this is challenging due to fluorophores’ spectral overlap and variation of the emission spectra. Here, we propose a strategy termed PICASSO, which enables more than 15-colour multiplexed imaging of thick tissue slices through a single imaging process and blind unmixing without reference spectra measurement. We show that PICASSO can be used to achieve a high multiplexing capability in diverse applications, such as 3D protein imaging, expansion microscopy, tissue clearing, imaging of clinical specimens, and cyclic immunofluorescence imaging. PICASSO only requires an equal number of images as the number of fluorophores, enabling such a high level of multiplexed imaging even with bandpass filter-based microscopy. As such, PICASSO would be a useful tool for the study of cancer, the immune system, and the brain, as well as for the diagnosis of cancer, as it enables ultra-multiplexed imaging of diverse specimens with minimum instrumental requirements and experimental processes.

Competing Interest Statement

J.-B.C., Y.-G.Y., J.S., Y.S. H.K., J.K., have applied for patents on PICASSO (KR patent application 10-2020-0088091, 10-2020-0106838, 10-2020-0109519, and US patent application 17/132,628).

Footnotes

  • ↵* e-mail: ygyoon{at}kaist.ac.kr; jbchang03{at}kaist.ac.kr

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted January 27, 2021.
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PICASSO: Ultra-multiplexed fluorescence imaging of biomolecules through single-round imaging and blind source unmixing
Junyoung Seo, Yeonbo Sim, Jeewon Kim, Hyunwoo Kim, In Cho, Young-Gyu Yoon, Jae-Byum Chang
bioRxiv 2021.01.27.428247; doi: https://doi.org/10.1101/2021.01.27.428247
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PICASSO: Ultra-multiplexed fluorescence imaging of biomolecules through single-round imaging and blind source unmixing
Junyoung Seo, Yeonbo Sim, Jeewon Kim, Hyunwoo Kim, In Cho, Young-Gyu Yoon, Jae-Byum Chang
bioRxiv 2021.01.27.428247; doi: https://doi.org/10.1101/2021.01.27.428247

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