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Related Experiment Video

Updated: Nov 12, 2025

In vivo Dual Substrate Bioluminescent Imaging
07:33

In vivo Dual Substrate Bioluminescent Imaging

Published on: October 11, 2011

15.8K

Rapid Multicomponent Bioluminescence Imaging via Substrate Unmixing.

Colin M Rathbun, Anastasia A Ionkina, Zi Yao

    ACS Chemical Biology
    |March 17, 2021
    PubMed
    Summary

    Researchers developed a new method for multiplexed bioluminescence imaging. This technique allows for the simultaneous detection of multiple bioluminescent reporters in real-time, overcoming previous limitations in tracking complex biological processes.

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    Fluorescent Protein Chromophore-Based Luciferins for Bioluminescence Imaging.

    Chemical & biomedical imaging·2025

    Area of Science:

    • Biotechnology
    • Molecular Imaging
    • Biochemistry

    Background:

    • Bioluminescence imaging offers sensitive, real-time monitoring of biological processes without external excitation light.
    • Existing methods struggle with simultaneous tracking of multiple bioluminescent reporters, hindering the study of complex cellular dynamics.
    • Engineered luciferases and luciferins have advanced bioluminescence imaging capabilities.

    Purpose of the Study:

    • To develop a streamlined strategy for rapid, multiplexed imaging using various luciferases and luciferins.
    • To enable simultaneous visualization of multiple bioluminescent reporters in vitro and in vivo.
    • To overcome limitations in capturing short-term changes in cell growth or gene expression.

    Main Methods:

    • Sequential addition of orthogonal luciferins to biological samples.

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    Last Updated: Nov 12, 2025

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  • Substrate unmixing techniques to differentiate signals from multiple luciferases.
  • Application of the method for in vitro and in vivo imaging, including in mice.

    • Main Results:

    • Demonstrated facile detection of multiple luciferases using the developed sequential substrate addition and unmixing strategy.
    • Achieved multicomponent bioluminescence imaging in vitro and in vivo.
    • Successfully performed multicomponent imaging in mice on a minutes-to-hours timescale.

    Conclusions:

    • The reported strategy enables rapid, multiplexed bioluminescence imaging with a broad range of luciferases and luciferins.
    • This approach facilitates the simultaneous tracking of multiple biological reporters, advancing real-time biological studies.
    • The method provides a valuable tool for investigating dynamic biological processes in complex systems.