<p>Multicolor bioluminescence imaging using near-infrared emitting luciferases is an attractive application to detect two cell populations within one animal model. Herein, we describe how to distinguish dual-color bioluminescent signals co-localized in the same compartment. We tested CBG2 click beetle (λ = 660 nm) and CBR2 click beetle (λ = 730 nm) luciferases paired with NH<sub>2</sub>-NpLH2 luciferin. Following a spectral unmixing algorithm, single spectral contributions can be resolved and quantified, enabling the visualization of multiple cell types in deep tissue by injection of a single substrate. For complete details on the use and execution of this protocol, please refer to Zambito et al. (2020).</p>

doi.org/10.1016/j.xpro.2021.100662, hdl.handle.net/1765/135785
STAR Protocols
Erasmus MC: University Medical Center Rotterdam

G. (Giorgia) Zambito, & L. (Laura) Mezzanotte. (2021). Near-infrared bioluminescence imaging of two cell populations in living mice. STAR Protocols, 2(3). doi:10.1016/j.xpro.2021.100662