Erasmus Medical Center, Dept. of Radiology and Nuclear Medicine, Rotterdam 3015 CE, the Netherlands.
Erasmus Medical Center, Dept. of Molecular Genetics, Rotterdam 3015 CE, the Netherlands.
STAR Protoc. 2021 Jul 7;2(3):100662. doi: 10.1016/j.xpro.2021.100662. eCollection 2021 Sep 17.
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-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).
使用近红外发射荧光素酶进行多色生物发光成像是一种很有吸引力的应用，可以在一个动物模型中检测两种细胞群体。在此，我们描述了如何区分同一隔室内共定位的双色生物发光信号。我们测试了 CBG2 叩头虫（λ=660nm）和 CBR2 叩头虫（λ=730nm）荧光素酶与 NH-NpLH2 荧光素的配对。通过光谱解混算法，可以解析和量化单个光谱贡献，从而通过注射单个底物在深部组织中可视化多种细胞类型。有关该协议的使用和执行的完整详细信息，请参见 Zambito 等人（2020 年）。
