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Caged luciferins enable rapid multicomponent bioluminescence imaging.

Mariana X Navarro1, Caroline K Brennan1, Anna C Love1

  • 1Department of Chemistry, University of California, Irvine, Irvine, California, USA.

Photochemistry and Photobiology
|June 1, 2023
PubMed
Summary

Multiplexed bioluminescence imaging is now more accessible. This study introduces caged luciferins and uncaging enzymes for easier multi-component tracking, simplifying complex biological imaging.

Keywords:
bioluminescenceimagingluciferaseluciferinsubstrate unmixing

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Bioimaging

Background:

  • Bioluminescence imaging offers sensitive, time-resolved visualization of biological processes.
  • Multiplexed tracking has been historically limited by the scarcity of orthogonal luciferase-luciferin pairs.
  • Existing methods for multi-parameter imaging often involve complex syntheses and enzyme evolution.

Purpose of the Study:

  • To present a simplified strategy for multiplexed bioluminescence imaging.
  • To demonstrate the utility of caged luciferins and established uncaging enzymes for multi-component tracking.
  • To reduce the complexity and accessibility barriers for multicomponent bioluminescence applications.

Main Methods:

  • Utilized orthogonal caged luciferins and pre-existing uncaging enzymes.
  • Developed a linear unmixing algorithm to distinguish unique bioluminescent signals.
  • Applied the method for two- and three-component imaging on the minutes timescale.

Main Results:

  • Successfully achieved multiplexed bioluminescence imaging using caged luciferins.
  • Demonstrated the ability to distinguish signals from different components via linear unmixing.
  • Validated the use of caged luciferins with endogenous enzymes for combined studies.

Conclusions:

  • This approach provides a more accessible method for multicomponent bioluminescence imaging.
  • Caged luciferins and uncaging enzymes offer a streamlined alternative to complex probe generation.
  • The technique is readily adoptable by the scientific community for advanced bioimaging.