Near-infrared bioluminescence imaging of two cell populations in living mice

Giorgia Zambito; Laura Mezzanotte

doi:10.1016/j.xpro.2021.100662

Near-infrared bioluminescence imaging of two cell populations in living mice

Giorgia Zambito, Laura Mezzanotte^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

3 Citations (Scopus)

Abstract

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 NH₂-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).

Original language	English
Article number	100662
Journal	STAR Protocols
Volume	2
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2021.100662
Publication status	Published - 17 Sept 2021

Bibliographical note

Funding Information:
We acknowledge the funding for this work provided by the European Commission under the H2020 -MSCA-RISE award grant number 777682 (CANCER). The graphical abstract and figures were partially created with BioRender.com .

Publisher Copyright:
© 2021 The Author(s)

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10.1016/j.xpro.2021.100662

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title = "Near-infrared bioluminescence imaging of two cell populations in living mice",

abstract = "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 NH2-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).",

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AU - Mezzanotte, Laura

N1 - Funding Information: We acknowledge the funding for this work provided by the European Commission under the H2020 -MSCA-RISE award grant number 777682 (CANCER). The graphical abstract and figures were partially created with BioRender.com . Publisher Copyright: © 2021 The Author(s)

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AB - 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 NH2-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).

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Near-infrared bioluminescence imaging of two cell populations in living mice

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