Journal
ISCIENCE
Volume 24, Issue 1, Pages -Publisher
CELL PRESS
DOI: 10.1016/j.isci.2020.101986
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Funding
- European Commission under the H2020-MSCARISE award [777682]
- European Commission under H2020-MSCA-ITN award [675743]
- Applied Molecular Imaging Erasmus MC (AMIE) facility
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In this study, a new click beetle luciferase mutant, CBG2, with red-shifted color emission is developed for in vivo multicolor bioluminescence applications. Paired with NH2-NpLH2 luciferin, CBG2 (λ=660nm) and CBR2 (λ=730nm) luciferases can be used for simultaneous dual-color bioluminescence imaging in deep tissue. A spectral unmixing algorithm tool can distinguish each spectral contribution, expanding the near-infrared bioluminescent toolbox for visualization of multiple biological processes in deep tissue.
For in vivo multicolor bioluminescence applications, red and near-infrared signals are desirable over shorter wavelength signals because they are not as susceptible to light attenuation by blood and tissue. Herein, we describe the development of a new click beetle luciferase mutant, CBG2, with a red-shifted color emission. When paired with NH2-NpLH2 luciferin, CBG2 (lambda = 660 nm) and CBR2 (lambda = 730 nm) luciferases can be used for simultaneous dual-color bioluminescence imaging in deep tissue. Using a spectral unmixing algorithm tool it is possible to distinguish each spectral contribution. Ultimately, this enzyme pair can expand the near-infrared bioluminescent toolbox to enable rapid visualization of multiple biological processes in deep tissue using a single substrate.
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