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A novel fluorescence lifetime imaging system that optimizes photon efficiency.

Ryan A Colyer¹, Claudia Lee, Enrico Gratton

¹Department of Biomedical Engineering, University of California, Irvine, USA.

Microscopy Research and Technique

|November 17, 2007

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We developed a low-cost digital frequency domain fluorescence lifetime imaging (DFD-FLIM) system. This system offers high-quality lifetime resolution comparable to existing methods, simplifying FLIM data acquisition and analysis.

Area of Science:

Microscopy
Biophysics
Photonics

Background:

Fluorescence lifetime imaging (FLIM) provides contrast in biological systems based on molecular properties.
High equipment cost and complex data analysis limit FLIM adoption.

Purpose of the Study:

To present a low-cost digital frequency domain FLIM (DFD-FLIM) system.
To enable FLIM with quality comparable to established methods.

Main Methods:

Developed a mathematical model for DFD-FLIM.
Physically implemented a low-cost DFD-FLIM system.
Performed error analysis to optimize lifetime acquisition.

Main Results:

The DFD-FLIM system provides lifetime resolution comparable to time-correlated single photon counting.

Related Experiment Videos

The implementation natively outputs data as phasors.

Error analysis identified parameters for maximizing lifetime acquisition quality.

Conclusions:

The DFD-FLIM method simplifies FLIM hardware and software.
It offers a new interface for data display and interpretation.
Optimizes accuracy in determining fluorescence lifetimes.