FLIM Playground: An interactive, end-to-end graphical user interface for analyzing single cells with fluorescence...

Area of Science:

• Biomedical Imaging
• Cellular Biology
• Data Science

Background:

• Fluorescence lifetime imaging microscopy (FLIM) is a powerful tool for cellular imaging.
• Current FLIM analysis involves fragmented, multi-step processes, hindering efficient data interpretation.
• A unified platform is needed to streamline FLIM data extraction and analysis.

Purpose of the Study:

• To introduce FLIM Playground, an interactive graphical platform for unified single-cell FLIM workflows.
• To provide a user-friendly, open-source solution for FLIM data extraction, analysis, and visualization.
• To accelerate biological insights and promote reproducibility in FLIM studies.

Main Methods:

• Developed FLIM Playground as an interactive, open-source Python application.
• Integrated data extraction with metadata checks, calibration, and feature extraction (lifetime, morphology, texture).
• Implemented real-time visual analytic modules for data analysis, including fitting and phasor methods.

Main Results:

• FLIM Playground successfully unifies single-cell FLIM workflows from data extraction to analysis.
• Lifetime extraction methods (fitting, phasor) were validated against commercial software and published data.
• Demonstrated biological insights from cancer cell line FLIM data, showcasing the platform's utility.

Conclusions:

• FLIM Playground significantly accelerates hypothesis-driven discovery in FLIM research.
• The platform enhances reproducibility through integrated user checks and standardized workflows.
• Its modular design supports future integration of new imaging modalities and analysis techniques.