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

STAR Protoc. 2021 Jul 7;2(3):100662. doi: 10.1016/j.xpro.2021.100662. eCollection 2021 Sep 17.

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).

Keywords: Biotechnology and bioengineering; Model Organisms; Molecular/Chemical Probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Cell Tracking / methods*
  • Coleoptera / enzymology
  • Female
  • Luciferases / analysis
  • Luciferases / chemistry
  • Luciferases / metabolism
  • Luciferins / analysis
  • Luciferins / chemistry
  • Luciferins / metabolism
  • Luminescent Measurements / methods*
  • Mice
  • Mice, Nude
  • Spectroscopy, Near-Infrared / methods*

Substances

  • Luciferins
  • Luciferases