FLUTE: A Python GUI for interactive phasor analysis of FLIM data

Area of Science:

• Biophotonics and imaging science
• Computational biology and bioinformatics

Background:

• Fluorescence lifetime imaging microscopy (FLIM) provides insights into fluorophore microenvironments.
• The fit-free phasor approach simplifies FLIM data interpretation.
• A lack of open-source Python GUIs hinders widespread adoption of phasor analysis for FLIM data.

Purpose of the Study:

• To introduce Fluorescence Lifetime Ultimate Explorer (FLUTE), an open-source Python GUI for phasor analysis of FLIM data.
• To address the need for user-friendly tools in FLIM data analysis.
• To facilitate broader application of phasor analysis in biomedical research.

Main Methods:

• Development of a Python-based graphical user interface (GUI) named FLUTE.
• Implementation of automated FLIM data calibration and interactive phasor plot exploration.
• Integration of simultaneous display of phasor plots and FLIM images with lifetime contrasts.
• Inclusion of functionality for calculating distances to known molecular species and data export.

Main Results:

• FLUTE provides a user-friendly platform for phasor analysis of time-domain FLIM data.
• The GUI simplifies complex FLIM data processing, including calibration and filtering.
• Tested with diverse datasets, including zebrafish embryos and in vitro cells, demonstrating robustness.
• Facilitates interactive visualization and analysis of large FLIM datasets.

Conclusions:

• FLUTE democratizes advanced FLIM phasor analysis for researchers without specialized expertise.
• The tool accelerates FLIM data analysis through automation and interactive visualization.
• FLUTE enhances the accessibility and utility of phasor-based FLIM analysis in biomedical research.