Real-time chemiluminescence imaging using nano-lantern probes.

Area of Science:

• Biophotonics
• Molecular Imaging
• Live-cell Imaging

Background:

• Chemiluminescence imaging offers excitation-free visualization from single cells to whole bodies.
• Traditional methods suffer from low temporal resolution due to weak signals and long exposure times.
• Recent development of highly luminescent Nano-lantern proteins offers a potential solution.

Purpose of the Study:

• To introduce Nano-lantern probes for high temporal resolution chemiluminescence imaging.
• To describe protocols for real-time imaging of living samples, including freely moving mice and optogenetic applications.
• To demonstrate the capability of Nano-lanterns for live imaging at video rates.

Main Methods:

• Utilized Nano-lantern and Nano-lantern-based functional indicators for enhanced chemiluminescence.
• Developed protocols for imaging Nano-lantern expressing tumor cells in freely moving mice.
• Integrated Nano-lantern calcium indicators with optogenetic tools for simultaneous observation.
• Synchronized illumination and camera acquisition for real-time imaging.

Main Results:

• Nano-lantern probes significantly improve temporal resolution in chemiluminescence imaging.
• Achieved real-time chemiluminescence imaging of living samples, including cells, plants, and small animals, at video rates.
• Successfully demonstrated imaging of Nano-lantern expressing tumor cells in a moving mouse.
• Showcased compatible use of Nano-lanterns with optogenetic tools.

Conclusions:

• Nano-lantern probes represent a significant advancement in chemiluminescence imaging technology.
• Enables high-temporal-resolution, real-time imaging of dynamic biological processes in vivo.
• Opens new avenues for studying biological functions in living systems with unprecedented detail.