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Spectral phasor analysis enables multiplexed microscopy with bioluminescent probes

Bioluminescent phasor is a new technology for multiplexed, excitation-free imaging at the microscale using luciferase–luciferin pairs. This platform can readily unmix the broad, overlapping emission spectra of bioluminescent reporters, making possible the dynamic tracking of cellular and molecular features over prolonged time periods.

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Fig. 1: Bioluminescent phasor.

References

  1. Suzuki, K. et al. Five colour variants of bright luminescent protein for real-time multicolour bioimaging. Nat. Commun. 7, 13718 (2016). This paper discusses a series of ultrabright reporters for bioluminescence microscopy.

    CAS  Article  Google Scholar 

  2. Fereidouni, F., Bader, A. N. & Gerritsen, H. C. Spectral phasor analysis allows rapid and reliable unmixing of fluorescence microscopy spectral images. Opt. Express 20, 12729–12741 (2012). This paper introduces spectral phasor analysis for unmixing fluorescence spectra.

    CAS  Article  Google Scholar 

  3. Cutrale, F. et al. Hyperspectral phasor analysis enables multiplexed 5D in vivo imaging. Nat. Methods 14, 149–152 (2017). This paper showcases rapid unmixing of multiple spectrally overlapping fluorophores via spectral phasor analysis.

    CAS  Article  Google Scholar 

  4. Dvornikov, A. & Gratton, E. Hyperspectral imaging in highly scattering media by the spectral phasor approach using two filters. Biomed. Opt. Express 9, 3503–3511 (2018). This paper demonstrates a streamlined, inexpensive setup for light collection that is relevant to spectral phasor analysis.

    CAS  Article  Google Scholar 

  5. Yeh, H. W. & Ai, H. W. Development and applications of bioluminescent and chemiluminescent reporters and biosensors. Annu. Rev. Anal. Chem. 12, 129–150 (2019). This Review article highlights bioluminescent biosensors, including various BRET-based reporters.

    CAS  Article  Google Scholar 

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This is a summary of: Yao, Z. et al. Multiplexed bioluminescence microscopy via phasor analysis. Nat. Methods https://doi.org/10.1038/s41592-022-01529-9 (2022)

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Spectral phasor analysis enables multiplexed microscopy with bioluminescent probes. Nat Methods 19, 801–802 (2022). https://doi.org/10.1038/s41592-022-01530-2

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