How to build a time-gated luminescence microscope

Area of Science:

• Biophotonics
• Microscopy
• Optical Imaging

Background:

• Autofluorescence limits sensitivity in filter-based fluorescence microscopy.
• Time-gated detection offers a practical solution to suppress autofluorescence.
• Recent advances have led to robust time-gated luminescence microscopes.

Purpose of the Study:

• To provide detailed protocols for modifying standard fluorescence microscopes.
• To enable time-gated luminescence imaging capabilities.
• To improve contrast and sensitivity in fluorescence microscopy.

Main Methods:

• Modification of standard fluorescence microscopes.
• Implementation of time-gated detection principles.
• Step-by-step protocol development for practical application.

Main Results:

• Successful adaptation of standard microscopes for time-gated luminescence imaging.
• Demonstrated improvement in contrast and sensitivity.
• Provided reproducible protocols for researchers.

Conclusions:

• Time-gated detection is a viable method to overcome autofluorescence limitations.
• The provided protocols facilitate the adoption of time-gated luminescence microscopy.
• Enhanced imaging performance is achievable with modified standard microscopes.