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Multiplexed bioluminescence microscopy via phasor analysis

Zi Yao, Caroline K. Brennan, Lorenzo Scipioni, Hongtao Chen, Kevin Ng, Michelle A. Digman, Jennifer A. Prescher
doi: https://doi.org/10.1101/2021.06.18.448905
Zi Yao
1Departments of Chemistry, University of California, Irvine, California 92697, United States
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Caroline K. Brennan
1Departments of Chemistry, University of California, Irvine, California 92697, United States
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Lorenzo Scipioni
4Laboratory for Fluorescence Dynamics, Biomedical Engineering Department, University of California, Irvine, California 92697, United States
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Hongtao Chen
4Laboratory for Fluorescence Dynamics, Biomedical Engineering Department, University of California, Irvine, California 92697, United States
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Kevin Ng
3Pharmaceutical Sciences, University of California, Irvine, California 92697, United States
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Michelle A. Digman
4Laboratory for Fluorescence Dynamics, Biomedical Engineering Department, University of California, Irvine, California 92697, United States
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  • For correspondence: jpresche@uci.edu mdigman@uci.edu
Jennifer A. Prescher
1Departments of Chemistry, University of California, Irvine, California 92697, United States
2Molecular Biology & Chemistry, University of California, Irvine, California 92697, United States
3Pharmaceutical Sciences, University of California, Irvine, California 92697, United States
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  • For correspondence: jpresche@uci.edu mdigman@uci.edu
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ABSTRACT

Microscopic bioluminescence imaging has been historically challenging due to a lack of detection methods and easily resolved probes. Here we combine bioluminescence with phasor analysis, an optical method commonly used to distinguish spectrally similar fluorophores. Bioluminescent phasor enabled rapid differentiation of multiple luciferase reporters and resonance energy transfer processes. The merger of bioluminescence and phasor analysis provides a platform for routine, time-lapse imaging of collections of cellular features.

Competing Interest Statement

The authors have declared no competing interest.

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 June 18, 2021.
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Multiplexed bioluminescence microscopy via phasor analysis
Zi Yao, Caroline K. Brennan, Lorenzo Scipioni, Hongtao Chen, Kevin Ng, Michelle A. Digman, Jennifer A. Prescher
bioRxiv 2021.06.18.448905; doi: https://doi.org/10.1101/2021.06.18.448905
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Multiplexed bioluminescence microscopy via phasor analysis
Zi Yao, Caroline K. Brennan, Lorenzo Scipioni, Hongtao Chen, Kevin Ng, Michelle A. Digman, Jennifer A. Prescher
bioRxiv 2021.06.18.448905; doi: https://doi.org/10.1101/2021.06.18.448905

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