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Color-tunable bioluminescence imaging portfolio for cell imaging
Shota Tamaki1, Nobuo Kitada1, Masahiro Kiyama1
1Department of Engineering Science, Graduate School of Informatics and Engineering, The University of Electro-Communications, Chofu, Tokyo, 182-8585, Japan.
Scientific Reports
|January 27, 2021
Summary
Researchers developed a novel color-tunable bioluminescence (BL) imaging system using coelenterazine (CTZ) analogues. This system enables multicolor imaging of molecular events in cells by producing diverse BL hues from blue to far-red.
Area of Science:
- Biochemistry
- Molecular Biology
- Biophotonics
Background:
- Bioluminescence (BL) imaging is a powerful tool for studying molecular and cellular processes.
- Existing BL systems often have limited spectral diversity, hindering multicolor applications.
- Coelenterazine (CTZ) and its analogues are crucial substrates for marine luciferases, influencing BL emission spectra.
Purpose of the Study:
- To develop a color-tunable bioluminescence imaging system with a broad spectral range.
- To synthesize and characterize novel CTZ analogues for enhanced BL properties.
- To investigate the spectral characteristics and luciferase specificity of new CTZ analogues.
Main Methods:
- Synthesis of twelve novel coelenterazine (CTZ) analogues.
- Evaluation of bioluminescence (BL) properties with marine luciferases (e.g., Renilla luciferase 8, Artificial Luciferase 16, NanoLuc).
- Characterization of optical properties including kinetic parameters, dose dependency, and luciferase specificity.
- Density Functional Theory (DFT) calculations to understand the theoretical basis of spectral shifts.
Main Results:
- Three groups of CTZ analogues produced diverse bioluminescence hues from blue to far-red.
- Specific analogues (e.g., 1d) induced significant spectral red-shifts (187 nm with RLuc8, 105 nm with ALuc16).
- CTZ analogues exhibited distinct luciferase specificities, with some selective for ALucs or NanoLuc.
Conclusions:
- A novel, color-tunable bioluminescence imaging system was established using new CTZ analogues.
- The system offers a comprehensive color palette for multicolor imaging applications.
- This technology enables efficient imaging of molecular events in mammalian cells with enhanced spectral resolution.