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How to build a time-gated luminescence microscope.

Dayong Jin1, Yiqing Lu1, Robert C Leif2

  • 1Advanced Cytometry Laboratories, MQ BioFocus Research Centre & Photonics Research Centre, Macquarie University, New South Wales, Australia.

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Summary

Time-gated detection suppresses autofluorescence in microscopy, enhancing sensitivity and contrast. This study provides protocols for adapting standard microscopes for time-gated luminescence imaging, improving fluorescence microscopy performance.

Keywords:
autofluorescencelanthanidemicroscopytime-gated luminescence

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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.